Can anyone make sense of this to explain to a layperson?

September 13, 2018

I'm trying to 'finalize' things.

As for that direct taper, I recall how that used to be many years ago when I was healthy and able to taper normally.

During my first real taper from 4 mg (single dose) I ran into an issue when the dose went below 2 mg. Aside from anything else, I did get strong hormonal reactions which manifested in different ways, including a skin disorder. Pretty muc everytime I hit a lower dose, I had to stop working out in the gym (for me, that included running). Intuitively or otherwise, I knew I had to be more active at lower levels of intensity, for a longer time. I even made up activities.

I can't replicate what happened in 2010/2011 etc. It's basically that bad that my sleep is usually fragmented, I may fall asleep again due to exhaustion or some other reason, and some sleep late morning/early afternoon gives me energy to get through the day !

Consideration: direct forced taper, single dose. Complicated. A long time ago, after that lorazepam I tried a small cut. The most obvious effect was reduced functioning/ much slower passing of time during the day. That's a simple effect. Basically, I'd have to get through the day on that one dose. There is underlying damage in the CNS, damage to the body. I'm inclined to think and worried that such a taper would just cause further damage. I'm no longer 'active' or healthy. So I wonder if that makes sense. (exposing damage covered up by clonazepam, and causing further damage)

I'm very hesitant to introduce a new messy benzo like Librium or diazepam, I'd never have done that till I let that GP get to me and put me on lorazepam.I can't taper on the basis that I'm fine, or all benzos are the same. Clonazepam is unique. And in some unique and dysfucntional way, clonazepam helps me sleep. Possibly by increasing the allostatic load ... (or similar)

Does it make any sense ? I'm a bit at a loss. I was never good with pills. I'd give my left arm and leg to go back to 2013 and do a plain C/T. (I know the drugs/health/aging problems are kiling me, albeit very slowly)

September 13, 2018

Liberty, I would give a leg and arm as well. For crying out loud, this experience has cost me my hair anyway!!!!

I accept that at some point I cannot figure everything out. I am trying to, I have tried to. It is in my nature to continue to do so but I think that the fear of not understanding maybe has kept me from going forward in some ways.

I'm trying to keep my plasma levels stable but am now in tolerance. I could taper were it not for the overwhelming and never-ending symptoms. They just won't stop. A few others have spoken of difficulty with gabapentin at low doses. But frankly, I think it's more of a duality in that it exposes the unresolved ativan damage as well as the ongoing influence of gabapentin.

I think I have figured out one of my issues with gabapentin and it is really simple in retrospect. I continue to work, albeit with significant difficulty. On some days, when the gabapentin has been more effective, I find it induces sleepiness during the day. Of course, I've been at work and had to fight that. I believe I was causing my CNS to upregulate the excitatory system as it fought the gabapentin's somnolence side-effect. Further, after a day or two of fighting daytime drowsiness, it would reduce somewhat and then I would end up with heightened withdrawal symptoms.

It is so simple, but I was chasing information so much I could not see the forest for the trees. I also think this was a significant issue with Ativan which I took 3 or 4 times during the day (out of the 11 original days I took it) when I had highly stressful issues to manage.

I am doing everything by the rules right now but I do not seem to be able to go forward so that is very, very frustrating.

Tapering has essentially become impossible, the pain just does not resolve; even with 1 micro taper of 0.5% I will be in agony that does not remit for days and days. My guess is that enough of my circuits are dysfunctional as to essentially keep me chronically hyperexcited.

-RST

September 14, 2018

dm123,

Do you have a definition of 'circuit' in this context ?

And I get the feeling that getting off this drug would increase my allostatic load ...

I would start here.

https://canvas.brown.edu/courses/851434/pages/neural-circuits

There’s hundreds of links of source material in this area online.

If you want to understand neural circuit dynamics you will have to study the basics like cable theory, neural oscillators , Central pattern generators (CPGs), etc.

Then google Eve Marder and her work on the STNS (stomatogastric nervous system). You can find excellent videos and papers from her.

This will get you started.

THen once you get through some of the basics you can restudy some specific neural circuits

Indirect and direct MSN motor circuits.

Voluntary motor circuits

Reflex motor circuit

ENS which deals with the digestive nervous system. This is independent of the CNS but also affected by the sympathetic and parasympathetic nervous systems, which are dependent on the CNS. So all these circuits are connected at an even higher level.

Hippocampal nervous system

Limbic circuits which affect the parasympathetic and sympathetic nervous systems which affect the ENS, etc, etc..

Once you do this you will understand how the balance of excitatory and inhibitory signaling is the yin and yang of the biological universe, with homeostasis as The overriding goal. When this balance is disrupted or distorted you become symptomatic.

Lots of good stuff.

September 14, 2018

Liberty, I would give a leg and arm as well. For crying out loud, this experience has cost me my hair anyway!!!!

I accept that at some point I cannot figure everything out. I am trying to, I have tried to. It is in my nature to continue to do so but I think that the fear of not understanding maybe has kept me from going forward in some ways.

I'm trying to keep my plasma levels stable but am now in tolerance. I could taper were it not for the overwhelming and never-ending symptoms. They just won't stop. A few others have spoken of difficulty with gabapentin at low doses. But frankly, I think it's more of a duality in that it exposes the unresolved ativan damage as well as the ongoing influence of gabapentin.

I think I have figured out one of my issues with gabapentin and it is really simple in retrospect. I continue to work, albeit with significant difficulty. On some days, when the gabapentin has been more effective, I find it induces sleepiness during the day. Of course, I've been at work and had to fight that. I believe I was causing my CNS to upregulate the excitatory system as it fought the gabapentin's somnolence side-effect. Further, after a day or two of fighting daytime drowsiness, it would reduce somewhat and then I would end up with heightened withdrawal symptoms.

It is so simple, but I was chasing information so much I could not see the forest for the trees. I also think this was a significant issue with Ativan which I took 3 or 4 times during the day (out of the 11 original days I took it) when I had highly stressful issues to manage.

I am doing everything by the rules right now but I do not seem to be able to go forward so that is very, very frustrating.
Tapering has essentially become impossible, the pain just does not resolve; even with 1 micro taper of 0.5% I will be in agony that does not remit for days and days. My guess is that enough of my circuits are dysfunctional as to essentially keep me chronically hyperexcited.

-RST

Stress always keeps popping up as a recurring theme.

I also understand the impatience. Healing is often times very slow.

I agree with the theory overload. I had to figure it out just so that i could understand how this could happen in the human body. However, i agree, at some point you just have to jump into the taper, crossover, or do whatever you need to do, and hope it works out.

We never know how any therapy will work out until we try it. Models are good, but never always correct when it comes to the human body. We are all different in how we response to thereapies.

If there’s one theme that i hope comes through on this thread, it’s that it goes way way beyond the GABAaR itself and those subunits. At a higher level none of this matters. Hence we needed a model to explain why or how the symptoms could persist long after washout......

September 14, 2018

dm123,

Lots of stuff.

Quick question. When you stated earlier that you believed that the damage caused by the lorazepam on the receptor level would have healed already (my choice of words), and you mentioned that the real issue is elsewhere (I'm not so sure), you mentioned the word 'circuit'.

I think that basically refers to the ways neurons interact with other neurons ? Is this an accurate albeit simplistic summarization ?

I can image that the 'circuits' have been disrupted. And as an analogy of someone who suffered a more 'conventional' brain injury, 'revalidation' (is that word correct in common English medical context?) would be required.

So some kind of revalidation would be required, most likely informal (without medical supervision). My take is that I'd have be be functioning reasonably well (can't completly forego sleep), and that I'd have to rebuild neural networks through (repetitive) activity.

Does that make any sense ?

That seems daunting, given everything that has happened and is occurring.

September 14, 2018

dm 123 I try to limit my questions to you , but as I recall you did taper on Librium? I just read another post that seemed to indicate Librium worked really well for them. Do you think it is an underrated option

September 15, 2018

dm123,

Lots of stuff.

Quick question. When you stated earlier that you believed that the damage caused by the lorazepam on the receptor level would have healed already (my chocie of words), and you mentioned that the real issue is elsewhere (I'm not so sure), you mentioned the word 'circuit'.

I think that basically refers to the ways neurons interact with other neurons ? Is this an accurate albeit simplistic summarization ?
I can image that the 'circuits' have been disrupted. And as an analogy of someone who suffered a more 'conventional' brain injury, 'revalidation' (is that word correct in common English medical context?) would be required.

So some kind of revalidation would be required, most likely informal (without medical supervision). My take is that I'd have be be functioning reasonably well (can't completly forego sleep), and that I'd have to rebuild neural networks through (repetitive) activity.

Does that make any sense ?

That seems daunting, given everything that has happened and is occurring.

Hi,

I think you meant ‘rehabilitation’ perhaps.

Yes, please research the topic emergent level of neural circuits. You will find tons of information on the topic. It’s a very very new area of research, and the tools that are required to clinically observe such behavior of neural circuits are decades away. However, they are slowly being developed. I will present some of the tools in a later Part. These tools will eventually show us how neural circuits truly behave at the higher level in real life biological entities. These tools are required to understand what happens to our neural circuits at the higher level when stressed. And as you know, by stressed, i mean pharmaceutical , emotional, physical, etc. So once the tools are developed, clinically they will be able to stress the test subject and observe how the circuits accomodate. Benzodiazaphine research would have to include controlled studies, with specific Benzodiazaphines, used in specifically controlled settings (dosages, steady state vs non steady state), in combination with these nanotools to discern the effect that they have on neural circuit homeostasis. They would then have to withdraw or taper a chronically exposed test subject, and observe how the circuit recovers. This is a long ways off.

So once these plastic changes of accomodation occur in the brain (think of the circuits as being adjusted to accomodate the stress), there are changes in many of the 6 pillars and in many of the component systems of the model that simply do not reverse when the insulted receptors (in this case the GABAaR) have long since “recovered” and the drug is washed out. These systems and pillars have a hard time adjusting back to “normal” because of reinforcing feedback loops as i describe in the next paragraph.

These plastic changes normally revert when the stress is removed, but stress removal does not occur sometimes to a sufficient degree during a long protracted withdrawal , tolerance, etc. Hormones are disrupted, neurotransmitters are off kilter (think in terms of the pillars), neurogeneisis is aberrant (this takes time to revert, especially in the hippocampus). So these changes become semi-permanent, and need the proper environment to revert. The GABAaR receptors may have healed, but these collateral systems are still a mess.

Proper environment obviously means slowly tapering off the benzo, “enriched environment” that I’ve discussed in the past, reduction in negative stress, proper rest and nutrition, moderate exercise, etc. Much of this was discussed a while back. I think that maintaining steady state is the easiest and most important thing one can do while on the benzo. It’s just my opinion based on the model that has been developed. As yoiu said, clonazepam has a pretty long half life, so perhaps that’s not as important for your recovery. Perhaps you need not crossover. But perhaps the onset of action or the way the clonazepam binds to the receptor (that’s very unique among benzos) might be what needs to be addressed for your case. How we address that, i do not know.

Also, tolerance, in and of itself, irrespective of half life, is a massive stressor on the neural circuitry.

Regarding “rehabilitation” , i don’t know how effective it will be if you are indeed in tolerance, because we aren’t removing the primary stressor on the circuits.

As I’ve said, once primary stressors are alleviated or at least moderated, the plastic changes can and do revert over time. But they need to be given the chance and the right environment. Neurotransmitters need to normalize, hormones (especially the stress hormones) need to normalize, etc.

I do think that your exercising is good, but just be careful, because your CNS cannot take much on the side of excitatbility. So moderate exercise is good. I know you have a good diet and most of your hormones are ok. They cannot test for neurotransmitter levels in brain (yet), so we don’t know where you are on that.

I hope this helps a bit.

September 15, 2018

Please see this excellent post from Terry on rTMS.

This is something that does affect neural circuit distortions like entrainment and hyperexcitability.

RST posted an article on this a long time back on this thread and there’s tons of studies on the therapeutic use of TMS for entrainment based vestibular dysfunction.

http://www.benzobuddies.org/forum/index.php?topic=209916.0

September 15, 2018

dm 123 I try to limit my questions to you , but as I recall you did taper on Librium? I just read another post that seemed to indicate Librium worked really well for them. Do you think it is an underrated option

Hi Matt,

I am in the last portions of the taper. Yes, i crossed over from assorted z drugs (ambien,lunesta) and lorazepam. I could not have tapered from them directly.

I do think it is a very underrated option, as i was just going to post a piece on this thread about the very same, but could not get to it due to lack of time.

My thoughts on the issue are this:

There are many people on BB who get ill from valium itself. By “many” we should take it into the proper context. Most people on BB are in pain and not doing well. So the population here is biased to the negative. So if we see 50% of the people here that get sick from valium, it is not representative of the real population in the outside world.

If we look at Ashton’s clinical studies that’s more representative of the effectiveness of valium as a crossover benzo for tapering. Here stats are very good (positive). I don’t have the time now to pull the stats but i am guessing around 90% success rate tapering off. Now that’s not to say that there are no relapses post taper. She might not have followed up. But the point is, there’s probably perhaps 10% of the people who crossover to Valium that get sick from valium itself.

So this is a very real lissue and from what I’ve read on this forum it can be very very disabiling.

They typically get sick when they take the drug, and sometimes moving to three doses a day does not help at all. So it’s more than just steady state.

I thought about it, and perhaps it’s due to valiums fast onset of action. It has a very fast onset of action. I’ve only taken it a few times, but it seems much more “powerful” than Librium in this respect. It’s like a sledghammer, to the librium’s hammer, if i were to describe it.

So librium has a very slow onset of action, and i would be curious to find out what would happen if some of the valium 3 times a day suffererers were to move to librium with 3 dosing a day.

So i do think it is an underrated option in this respect.

I hope this helps.

Best wishes.

Just wanted to add, that the taper has not been easy. Anything but. I don’t know if there is any easy way out of this, but perhaps librium might be an option for some who are doing worse on valium.

P.S. Sorry for typos if any, as i typed it very quick and did not have a chance to spellcheck.

September 15, 2018

dm123,

I was taking a look back at the six pillars, as described in your ´page guide´.

I got it a little messy without reading through every single page ...

Something like this: ´1. The action potential dynamics at the individual neuron level. This is directly affected by the stress system (via LTP and LTD), the glutamatergic system, the GABAergic system, as well as a number of “other systems” that have not been presented yet.

2. The neurogenesis component, which accounts, in part, for benzodiazaphine PWS. Neurogenesis initially can be destabilized by dysfunction in the GABAergic system (alcohol or benzodiazaphines; see the alcohol study referred to in an earlier post, one of many references in this area) and perpetuated through the negative stress system.Neurogenesis is directly affected by the glutamatergic, GABAergic and stress systems, as well as many other different neurogenic modulators (including serotonin) that were briefly discussed in the Neurogenesis (PART 1) post above.

3. The neuroplasticity overlay, which includes neural circuit dynamics, and homeostatic plasticity, as well as hebbian plasticity (LTP and LTD; note this is not the same as non-hebbian glutamatergic LTP which can be associated with PWS under abrupt withdrawal). Neuroplasticity is also affected by the stress system, the GABAergic system, and the glutamatergic system, as well as many many (the list is huge!)different neuromodulators.

The 3 major components above are affected by benzodiazaphines through the 3 major mechanistic systems or “pillars” alluded to above (reduced to 3 for simplicity):

1. The glutamatergic system,

2. the GABAergic system, and of course

3. the stress system.

ON/ONOO- cycles, Peroxynitrate, and Blood pressure ´

The last one isn´t entirely clear to me, and to be honest I´m not very interested/there´d be little I could do about that. Likely not my issue.

And there is the GCPR and the circuit issue ...

The latter two are reasonably clear to me.

Well, not looking good for me.

For me: GABA, GCPRs, hormones, circuit level, chronic stress, abberant neurogenesis, iatrogenic damage (outside the CNS) caused by other health problems that does not neatly fall into the category of ´disease´.

I think that clonazepam reconfigures the GCPRs, and that all those health problems, drug switches, interdose etc. geconfigured the GCPRs even more. I have been on this drug for a decade or so.

´I could not handle clonazepam. I rapidly developed tolerance within weeks.´ Not me. The 2 mg dose did last for less than a year, the full 4 mg a full year. Then I decided tapering was the wisest course of action. Rough, lots of problems. I made numerous mistakes. But what did I know ?

Since 2013 at about 2 mg. Till the lorazepam in 2015 it continued working, possibly a bit less.

In reality, health problems were the greater issue. That mysterious exercise factor, I could go lower if I abstained from exercise. A not so benzo like benzo. (antimyoclic, anti absence, anticonvulsant properties that extend beyond GABA) I know you were at a lower dose. I didn´t know how dangerous this drug really was.

Well, I´m a mess in a country that has an inferior healthcare system. Basically, the drug is no longer compatible with my body. And how to get a Dutch doctor to diagnose iatrogenic injuries ?

As for diazepam or Librium, there is always a risk in switching. While Librium acts relatively strong at alpha 3, diazepam has a very strong affinity for alpha 1 and 5. Librium low potency, diazepam medium potency. It´s very individual. Ashton never tried clonazepam.

September 16, 2018

dm123,

I was taking a look back at the six pillars, as described in your ´page guide´.

I got it a little messy without reading through every single page ...

Something like this: ´1. The action potential dynamics at the individual neuron level. This is directly affected by the stress system (via LTP and LTD), the glutamatergic system, the GABAergic system, as well as a number of “other systems” that have not been presented yet.

2. The neurogenesis component, which accounts, in part, for benzodiazaphine PWS. Neurogenesis initially can be destabilized by dysfunction in the GABAergic system (alcohol or benzodiazaphines; see the alcohol study referred to in an earlier post, one of many references in this area) and perpetuated through the negative stress system.Neurogenesis is directly affected by the glutamatergic, GABAergic and stress systems, as well as many other different neurogenic modulators (including serotonin) that were briefly discussed in the Neurogenesis (PART 1) post above.

3. The neuroplasticity overlay, which includes neural circuit dynamics, and homeostatic plasticity, as well as hebbian plasticity (LTP and LTD; note this is not the same as non-hebbian glutamatergic LTP which can be associated with PWS under abrupt withdrawal). Neuroplasticity is also affected by the stress system, the GABAergic system, and the glutamatergic system, as well as many many (the list is huge!)different neuromodulators.

The 3 major components above are affected by benzodiazaphines through the 3 major mechanistic systems or “pillars” alluded to above (reduced to 3 for simplicity):

1. The glutamatergic system,

2. the GABAergic system, and of course

3. the stress system.

ON/ONOO- cycles, Peroxynitrate, and Blood pressure ´
The last one isn´t entirely clear to me, and to be honest I´m not very interested/there´d be little I could do about that. Likely not my issue.

And there is the GCPR and the circuit issue ...

The latter two are reasonably clear to me.

Well, not looking good for me.

For me: GABA, GCPRs, hormones, circuit level, chronic stress, abberant neurogenesis, iatrogenic damage (outside the CNS) caused by other health problems that does not neatly fall into the category of ´disease´.
I think that clonazepam reconfigures the GCPRs, and that all those health problems, drug switches, interdose etc. geconfigured the GCPRs even more. I have been on this drug for a decade or so.

´I could not handle clonazepam. I rapidly developed tolerance within weeks.´ Not me. The 2 mg dose did last for less than a year, the full 4 mg a full year. Then I decided tapering was the wisest course of action. Rough, lots of problems. I made numerous mistakes. But what did I know ?
Since 2013 at about 2 mg. Till the lorazepam in 2015 it continued working, possibly a bit less.

In reality, health problems were the greater issue. That mysterious exercise factor, I could go lower if I abstained from exercise. A not so benzo like benzo. (antimyoclic, anti absence, anticonvulsant properties that extend beyond GABA) I know you were at a lower dose. I didn´t know how dangerous this drug really was.

Well, I´m a mess in a country that has an inferior healthcare system. Basically, the drug is no longer compatible with my body. And how to get a Dutch doctor to diagnose iatrogenic injuries ?

As for diazepam or Librium, there is always a risk in switching. While Librium acts relatively strong at alpha 3, diazepam has a very strong affinity for alpha 1 and 5. Librium low potency, diazepam medium potency. It´s very individual. Ashton never tried clonazepam.

Yes, I know, it is very overwhelming.

It is all very individual. One size does not fit all.

September 17, 2018

dm123,

I doubt anyone has ever collected and integrated this knowledge the way you have !

I do have a question though.

[/i]
'Section 2
Quote
Activity-dependent GABAAR ubiquitination The ER is responsible for the retention and degradation of misfolded or unassembled subunits and, accordingly, homomeric unassembled GABAARs subunits have been shown to be degraded in this organelle14,18. ER-associated degradation (ERAD) involves protein ubiquitination and degradation via the ubiquitin-proteasome system (UPS)19. GABAAR subunits have recently been shown to be ubiquitinated in an activity-dependent manner20. Chronic blockade of neuronal activity dramatically increased the levels of GABAAR ubiquitination within the ER, resulting in decreased insertion at the plasma membrane20. Correspondingly, increasing the level of neuronal activity resulted in a decrease in the level of GABAAR ubiquitination and an enhancement of receptor cell surface expression20. Thus, neuronal activity can regulate the ubiquitination of GABAARs in the ER, affecting their rate of degradation via the UPS. This may be one mechanism neurons use to homeostatically regulate synaptic inhibition.
End quote'

I'm having a very hard time making sense of this. I did look up 'ubiquitination'. This blockade of neuronal activity, what kind are we talking about ? Inactivity, drugs, taking benzodiazepines ?

Hi liberty, the blockade of neural activity is extreme, As is the way they caused neuronal activity. However, consistent with the theme of this thread, this is a grand example of another homeostatic response at the cellular level. I hope this is not too much info, but I wanted to answer your question in every way. This next ADDENDUM 7 could be thought of as being part of the downregulation bucket, item 3 in our GABAa list from ADDENDUM 6

I could not do much formatting on this, due to time, but here it is. If anyone knows of a way to easily import word docs with the formatting intact, let me know

This one is definitely worth reading.

ADDENDUM 7: cellular regulatory homeostasis in action: Chronic changes in neuronal activity regulate the numbers of GABAAR expressed on the neuronal cell surface

~"You gonna see, but you ain't gonna believe." - Sergio Oliva

Here is the source reference cited in that quote above. This is the full text. You have great questions. I had a similar question way back when I started researching this area. This study gives us incredible insight into a very complex cellular homeostatic response to external stimuli, via The level of Neuronal activity in the postsynaptic neuron.

http://www.jneurosci.org/content/27/48/13341.long

The 2011 reference goes into much further detail about the ubiquitination process. Section X, Part B, and it references the same footnote as that in the 2008 article that I quoted above. I read the section B and it’s quite confusing, but here is the relevant part. We can then work backwards.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4382539/#!po=22.1154

Quote1
The regulation of GABAAR trafficking by the ubiquitin-related protein Plic1 suggests that GABAARs may also be direct targets for modification by the polypeptide ubiquitin. The covalent attachment of one or more copies of the 76-amino acid ubiquitin monomer to lysine residues of target proteins is referred to as ubiquitination. Monoubiquitination is reversible and serves as an active signal in diverse intracellular trafficking pathways, including as a trigger for endocytosis. In contrast, polyubiquitination is required for the translocation of proteins from the ER back into the cytosol, where they are degraded by the proteasome. Activity-dependent polyubiquitination of GABAAR β3 subunits has been shown to reduce the stability of newly-translated and assembled receptors in the ER via a mechanism dependent on the activity of the proteasome (Saliba et al., 2007).
Coincident with a loss of cell surface expression levels, chronic blockade of neuronal activity by tetrodotoxin (TTX) treatment reduced both the amplitude
frequency of mIPSCs (Saliba et al., 2007). TTX had no effect on the enhanced functional expression of GABAARs incorporating β3 subunits in which all twelve lysine residues within the ICD of this subunit had been mutated to arginines (β3K12R). These mutations did not alter GABAAR cell surface half-life or internalization rates but did significantly enhance receptor insertion into the plasma membrane (Saliba et al., 2007).

End quote1

We can start by figuring out some terms.
First, in reading the very confusing quote 1 above, we want to focus on polyubiquitination.

I’m sure you read the ubiquitination article on Wikipedia.
The important part is quoted below

https://en.m.wikipedia.org/wiki/Ubiquitin

Quote2
The addition of ubiquitin to a substrate protein is called ubiquitination or less frequently ubiquitylation. Ubiquitination affects proteins in many ways: it can mark them for degradation via the proteasome, alter their cellular location, affect their activity, and promote or prevent protein interactions.[4][5][6] Ubiquitination involves three main steps: activation, conjugation, and ligation, performed by ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), and ubiquitin ligases (E3s), respectively. The result of this sequential cascade is to bind ubiquitin to lysine residues on the protein substrate via an isopeptide bond, cysteineresidues through a thioester bond, serine and threonine residues through an ester bond, or the amino group of the protein's N-terminus via a peptide bond.[7][8][9]
End quote2

The key here is binding to lysine residues on the target protein substrate by ubiquitin = ubiquitination
Our target protein for the purposes of the study referenced above(the original 2008 quote and quote1 above ) are the beta3 subunits of the GABAa receptors.
Next, we need to find out what this substance is called tetrodotoxin or TTX, because that’s what they are using to cause a chronic blockade of all neuronal activity.

As suspected TTX is a potent neurotoxin

https://en.m.wikipedia.org/wiki/Tetrodotoxin

This is where our knowledge presented in the previous ADDENDUMs on action potentials comes in
Here is what TTX does

Quote3

Tetrodotoxin is a sodium channel blocker. It inhibits the firing of action potentials in neurons by binding to the voltage-gated sodium channels in nerve cell membranes and blocking the passage of sodium ions (responsible for the rising phase of an action potential) into the neuron. This prevents the nervous system from carrying messages and thus muscles from flexing in response to nervous stimulation.[1]
Its mechanism of action, selective blocking of the sodium channel, was shown definitively in 1964 by Toshio Narahashi and John W. Moore at Duke University, using the sucrose gap voltage clamp technique.[2]
End quote3

It completely blocks the voltage gated sodium channels in the post synaptic neuron. This will completely prevent any current from going down the axon, because even if threshold potential is reached via the ion channel influx of Na+, the action potential or AP , for short, won’t be triggered because the sodium channels won’t be able to open to let all of the voltage gated sodium in required for that massive depolarization that is needed to create an AP.

So this is their definition of blocking neuronal activity. So when they did this in that 2007 study, nothing was getting through in terms of AP, and they found that this dramatically increased the ubiquitination of the GABAaR subunits in the Cell. This resulted in a loss of cell surface expression levels of the receptor. The end result was lower frequency and amplitude of mIPSCs or miniature inhibitory post synaptic currents. Not good

From the quote above “polyubiquitination is required for the translocation of proteins in the ER or endoplasmic reticulum back into the cytosol, where they are degraded by the proteasome…..”. The ER contains unassembled GABAa receptor subunits. These are targeted for ER-associated degradation by being ubiquitinated and degraded by the proteasome. A proteasome is a protein that degrades another protein that has been tagged by or bound by ubiquitin.

They found the opposite was true when neuronal activity was increased, i.e. Ubiquitination was decreased and there was an enhancement of receptor cell surface expression. More receptors made it to the surface.…. More about how they increased neuronal activity later. (Note : don’t confuse Plic-1 and 2 ubiquitin like proteins with ubiquitin)

To test the hypothesis that ubiquitination was involved and that there was a loss of cell surface expression of GABAaRs due to ubiquitination, they created mutated variants of GABAaRs where all the lysine residues in the beta3 unit were mutated to arginines. As explained above ,the ubiquitin cannot bind to the beta3 subunit without lysine present, thus it can’t ubiquitinate the receptor subunits . When they did that the TTX had no effect on the expression of these mutated receptors!!!!!
Note, These mutations did not alter the receptor half life or internalization rates but increased receptor insertion into the plasma membrane because ubiquitination was not occurring.
So , in summary and in layman’s terms, lack of neuronal activity via this neurotoxin that completely knocks out the AP from occurring, increases ubiquitination, i.e. Degradation , of these subunits inside the cell, thereby decreasing the receptor surface expression. The opposite, increasing neural activity did the opposite. This was confirmed by blocking ubiquitination via the mutated beta receptor. This is what they called “activity dependent ubiquitination “

So what did they do to increase neuronal activity? In going through the original study from 2007, see quote 6 below. They used a GABA antagonist. It was not flumazenil, but this might be why flumazenil therapy is so effective in tapering recovery! Amazing.

Using the body’s homeostatic response to our advantage by increasing neuronal stimulation and activity. Homeostatically, this all makes sense. You try to put the neuron to sleep by excessive inhibitory potentials, and the neuron responds by producing less GABAa receptors that make it to the surface, You excite and stimulate the neuron, as painful as it is, and the neuron responds by slowing the degradation of subunits (inside the cell)thereby increasing the number of assembled receptors that make it to the surface. This gives a whole new appreciation to the term downregulation, and gives us insight into one of the mechansims that the cell uses to achieve this.

So now the original study should make sense. Here are some quotes
And here are some main points. What an awesome study:
-Chronic changes in neuronal activity regulate the numbers of GABAAR expressed on the neuronal cell surface
-Chronic changes in neuronal activity regulate the levels of GABAAR β3-subunit ubiquitination
-Identification of major sites for ubiquitination within the GABAAR β3 subunit
- Decreasing β3 subunit ubiquitination increases receptor accumulation on the neuronal cell surface
- Ubiquitination of the β3 subunit modulates GABAAR insertion into the plasma membrane
- The effects of neuronal activity on GABAAR cell surface expression levels are mediated via β3 subunit ubiquitination

-Chronic blockade of neuronal activity regulates the accumulation of GABAARs containing β3 subunits at synaptic sites

Quote 4
Treatment of cultures with 2 μm TTX for 24 h resulted in a significant decrease (to 47.8 ± 5.3% of control) (Fig. 1A) in cell surface levels of GABAARs containing β3 subunits. The total expression of receptor β3 subunits was also reduced by this treatment (to 45 ± 6.7% of control) (Fig. 1A). Similar statistically significant decreases in the cell surface expression levels of GABAAR α2 and γ2 subunits were also evident (supplemental Fig. 1, available at www.jneurosci.org as supplemental material), suggesting that chronic neuronal inactivity leads to the loss of functional heteromeric cell surface GABAARs. In addition TTX treatment also significantly decreased the number of cell surface GABAARs containing β3 subunits in 16 DIV hippocampal neurons
End quote

This is really interesting, with Glutamate antagonists to reduce neuronal activity.

Quote 5
To corroborate our experiments with TTX, we assessed the effects of blocking excitatory synaptic transmission on the cell surface levels of GABAARs with glutamate receptor antagonists, which have been established to reduce neuronal activity in culture (Rao and Craig, 1997; O'Brien et al., 1998). Blockade of glutamate receptors significantly decreased the cell surface expression levels of GABAARs containing β3 subunits to 45.8 ± 13.1% of control (Fig. 1B)
End quote

They used a GABAa antagonist, PTX to increase neuronal activity,

Quote 6
We also examined the effects of chronically increasing neuronal activity on the cell surface expression levels of GABAARs using picrotoxin (PTX) (40 μm), a GABAAR antagonist and has been shown to increase the activity of neuronal cultures (Rao and Craig, 1997; O'Brien et al., 1998). In contrast to the effects of TTX and glutamate receptor antagonists, treatment of neurons with PTX significantly increased both the cell surface (by 55 ± 2.3% of control) and total (by 47 ± 8.5% of control) expression levels of the GABAARs β3 subunit (Fi
End quote

So now the abstract should make sense.....

Quote 7
Abstract
GABA(A) receptors (GABA(A)Rs) are the major mediators of fast synaptic inhibition in the brain. In neurons, these receptors undergo significant rates of endocytosis and exocytosis, processes that regulate both their accumulation at synaptic sites and the efficacy of synaptic inhibition. Here we have evaluated the role that neuronal activity plays in regulating the residence time of GABA(A)Rs on the plasma membrane and their targeting to synapses. Chronic blockade of neuronal activity dramatically increases the level of the GABA(A)R ubiquitination, decreasing their cell surface stability via a mechanism dependent on the activity of the proteasome. Coincident with this loss of cell surface expression levels, TTX treatment reduced both the amplitude and frequency of miniature inhibitory synaptic currents. Conversely, increasing the level of neuronal activity decreases GABA(A)R ubiquitination enhancing their stability on the plasma membrane. Activity-dependent ubiquitination primarily acts to reduce GABA(A)R stability within the endoplasmic reticulum and, thereby, their insertion into the plasma membrane and subsequent accumulation at synaptic sites. Thus, activity-dependent ubiquitination of GABA(A)Rs and their subsequent proteasomal degradation may represent a potent mechanism to regulate the efficacy of synaptic inhibition and may also contribute to homeostatic synaptic plasticity.
End quote

I took lysine 500mg last night after a particularly rough day after I read a post on the board where it helped with sleep. I slept 7 hours straight with no adrenaline surges last night. I had been sleeping 2-3 hours a night with atleast two adrenaline surges a night and none of this. I think by reading this post that lysine works on gaba receptors is that the take away? My benzo brain isn’t really comprehending what the article states.

September 17, 2018

dm123,

I'm sorry I don't remember all the theory.

For a while I was looking at a few things. It's true that not only gaba and glutamate are involved in the process of the action potential, but also neurotransmitters like acetylcholine and serotonin (I could provide links) ?

If that's true, then things like what we eat (serotonin), exercise/stress (acetylcholine) could also affect the action potential ? And thus GABA. You may have put it in different words ...

September 17, 2018

dm123,

I doubt anyone has ever collected and integrated this knowledge the way you have !

I do have a question though.

[/i]
'Section 2
Quote
Activity-dependent GABAAR ubiquitination The ER is responsible for the retention and degradation of misfolded or unassembled subunits and, accordingly, homomeric unassembled GABAARs subunits have been shown to be degraded in this organelle14,18. ER-associated degradation (ERAD) involves protein ubiquitination and degradation via the ubiquitin-proteasome system (UPS)19. GABAAR subunits have recently been shown to be ubiquitinated in an activity-dependent manner20. Chronic blockade of neuronal activity dramatically increased the levels of GABAAR ubiquitination within the ER, resulting in decreased insertion at the plasma membrane20. Correspondingly, increasing the level of neuronal activity resulted in a decrease in the level of GABAAR ubiquitination and an enhancement of receptor cell surface expression20. Thus, neuronal activity can regulate the ubiquitination of GABAARs in the ER, affecting their rate of degradation via the UPS. This may be one mechanism neurons use to homeostatically regulate synaptic inhibition.
End quote'

I'm having a very hard time making sense of this. I did look up 'ubiquitination'. This blockade of neuronal activity, what kind are we talking about ? Inactivity, drugs, taking benzodiazepines ?

Hi liberty, the blockade of neural activity is extreme, As is the way they caused neuronal activity. However, consistent with the theme of this thread, this is a grand example of another homeostatic response at the cellular level. I hope this is not too much info, but I wanted to answer your question in every way. This next ADDENDUM 7 could be thought of as being part of the downregulation bucket, item 3 in our GABAa list from ADDENDUM 6

I could not do much formatting on this, due to time, but here it is. If anyone knows of a way to easily import word docs with the formatting intact, let me know

This one is definitely worth reading.

ADDENDUM 7: cellular regulatory homeostasis in action: Chronic changes in neuronal activity regulate the numbers of GABAAR expressed on the neuronal cell surface

~"You gonna see, but you ain't gonna believe." - Sergio Oliva

Here is the source reference cited in that quote above. This is the full text. You have great questions. I had a similar question way back when I started researching this area. This study gives us incredible insight into a very complex cellular homeostatic response to external stimuli, via The level of Neuronal activity in the postsynaptic neuron.

http://www.jneurosci.org/content/27/48/13341.long

The 2011 reference goes into much further detail about the ubiquitination process. Section X, Part B, and it references the same footnote as that in the 2008 article that I quoted above. I read the section B and it’s quite confusing, but here is the relevant part. We can then work backwards.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4382539/#!po=22.1154

Quote1
The regulation of GABAAR trafficking by the ubiquitin-related protein Plic1 suggests that GABAARs may also be direct targets for modification by the polypeptide ubiquitin. The covalent attachment of one or more copies of the 76-amino acid ubiquitin monomer to lysine residues of target proteins is referred to as ubiquitination. Monoubiquitination is reversible and serves as an active signal in diverse intracellular trafficking pathways, including as a trigger for endocytosis. In contrast, polyubiquitination is required for the translocation of proteins from the ER back into the cytosol, where they are degraded by the proteasome. Activity-dependent polyubiquitination of GABAAR β3 subunits has been shown to reduce the stability of newly-translated and assembled receptors in the ER via a mechanism dependent on the activity of the proteasome (Saliba et al., 2007).
Coincident with a loss of cell surface expression levels, chronic blockade of neuronal activity by tetrodotoxin (TTX) treatment reduced both the amplitude
frequency of mIPSCs (Saliba et al., 2007). TTX had no effect on the enhanced functional expression of GABAARs incorporating β3 subunits in which all twelve lysine residues within the ICD of this subunit had been mutated to arginines (β3K12R). These mutations did not alter GABAAR cell surface half-life or internalization rates but did significantly enhance receptor insertion into the plasma membrane (Saliba et al., 2007).

End quote1

We can start by figuring out some terms.
First, in reading the very confusing quote 1 above, we want to focus on polyubiquitination.

I’m sure you read the ubiquitination article on Wikipedia.
The important part is quoted below

https://en.m.wikipedia.org/wiki/Ubiquitin

Quote2
The addition of ubiquitin to a substrate protein is called ubiquitination or less frequently ubiquitylation. Ubiquitination affects proteins in many ways: it can mark them for degradation via the proteasome, alter their cellular location, affect their activity, and promote or prevent protein interactions.[4][5][6] Ubiquitination involves three main steps: activation, conjugation, and ligation, performed by ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), and ubiquitin ligases (E3s), respectively. The result of this sequential cascade is to bind ubiquitin to lysine residues on the protein substrate via an isopeptide bond, cysteineresidues through a thioester bond, serine and threonine residues through an ester bond, or the amino group of the protein's N-terminus via a peptide bond.[7][8][9]
End quote2

The key here is binding to lysine residues on the target protein substrate by ubiquitin = ubiquitination
Our target protein for the purposes of the study referenced above(the original 2008 quote and quote1 above ) are the beta3 subunits of the GABAa receptors.
Next, we need to find out what this substance is called tetrodotoxin or TTX, because that’s what they are using to cause a chronic blockade of all neuronal activity.

As suspected TTX is a potent neurotoxin

https://en.m.wikipedia.org/wiki/Tetrodotoxin

This is where our knowledge presented in the previous ADDENDUMs on action potentials comes in
Here is what TTX does

Quote3

Tetrodotoxin is a sodium channel blocker. It inhibits the firing of action potentials in neurons by binding to the voltage-gated sodium channels in nerve cell membranes and blocking the passage of sodium ions (responsible for the rising phase of an action potential) into the neuron. This prevents the nervous system from carrying messages and thus muscles from flexing in response to nervous stimulation.[1]
Its mechanism of action, selective blocking of the sodium channel, was shown definitively in 1964 by Toshio Narahashi and John W. Moore at Duke University, using the sucrose gap voltage clamp technique.[2]
End quote3

It completely blocks the voltage gated sodium channels in the post synaptic neuron. This will completely prevent any current from going down the axon, because even if threshold potential is reached via the ion channel influx of Na+, the action potential or AP , for short, won’t be triggered because the sodium channels won’t be able to open to let all of the voltage gated sodium in required for that massive depolarization that is needed to create an AP.

So this is their definition of blocking neuronal activity. So when they did this in that 2007 study, nothing was getting through in terms of AP, and they found that this dramatically increased the ubiquitination of the GABAaR subunits in the Cell. This resulted in a loss of cell surface expression levels of the receptor. The end result was lower frequency and amplitude of mIPSCs or miniature inhibitory post synaptic currents. Not good

From the quote above “polyubiquitination is required for the translocation of proteins in the ER or endoplasmic reticulum back into the cytosol, where they are degraded by the proteasome…..”. The ER contains unassembled GABAa receptor subunits. These are targeted for ER-associated degradation by being ubiquitinated and degraded by the proteasome. A proteasome is a protein that degrades another protein that has been tagged by or bound by ubiquitin.

They found the opposite was true when neuronal activity was increased, i.e. Ubiquitination was decreased and there was an enhancement of receptor cell surface expression. More receptors made it to the surface.…. More about how they increased neuronal activity later. (Note : don’t confuse Plic-1 and 2 ubiquitin like proteins with ubiquitin)

To test the hypothesis that ubiquitination was involved and that there was a loss of cell surface expression of GABAaRs due to ubiquitination, they created mutated variants of GABAaRs where all the lysine residues in the beta3 unit were mutated to arginines. As explained above ,the ubiquitin cannot bind to the beta3 subunit without lysine present, thus it can’t ubiquitinate the receptor subunits . When they did that the TTX had no effect on the expression of these mutated receptors!!!!!
Note, These mutations did not alter the receptor half life or internalization rates but increased receptor insertion into the plasma membrane because ubiquitination was not occurring.
So , in summary and in layman’s terms, lack of neuronal activity via this neurotoxin that completely knocks out the AP from occurring, increases ubiquitination, i.e. Degradation , of these subunits inside the cell, thereby decreasing the receptor surface expression. The opposite, increasing neural activity did the opposite. This was confirmed by blocking ubiquitination via the mutated beta receptor. This is what they called “activity dependent ubiquitination “

So what did they do to increase neuronal activity? In going through the original study from 2007, see quote 6 below. They used a GABA antagonist. It was not flumazenil, but this might be why flumazenil therapy is so effective in tapering recovery! Amazing.

Using the body’s homeostatic response to our advantage by increasing neuronal stimulation and activity. Homeostatically, this all makes sense. You try to put the neuron to sleep by excessive inhibitory potentials, and the neuron responds by producing less GABAa receptors that make it to the surface, You excite and stimulate the neuron, as painful as it is, and the neuron responds by slowing the degradation of subunits (inside the cell)thereby increasing the number of assembled receptors that make it to the surface. This gives a whole new appreciation to the term downregulation, and gives us insight into one of the mechansims that the cell uses to achieve this.

So now the original study should make sense. Here are some quotes
And here are some main points. What an awesome study:
-Chronic changes in neuronal activity regulate the numbers of GABAAR expressed on the neuronal cell surface
-Chronic changes in neuronal activity regulate the levels of GABAAR β3-subunit ubiquitination
-Identification of major sites for ubiquitination within the GABAAR β3 subunit
- Decreasing β3 subunit ubiquitination increases receptor accumulation on the neuronal cell surface
- Ubiquitination of the β3 subunit modulates GABAAR insertion into the plasma membrane
- The effects of neuronal activity on GABAAR cell surface expression levels are mediated via β3 subunit ubiquitination

-Chronic blockade of neuronal activity regulates the accumulation of GABAARs containing β3 subunits at synaptic sites

Quote 4
Treatment of cultures with 2 μm TTX for 24 h resulted in a significant decrease (to 47.8 ± 5.3% of control) (Fig. 1A) in cell surface levels of GABAARs containing β3 subunits. The total expression of receptor β3 subunits was also reduced by this treatment (to 45 ± 6.7% of control) (Fig. 1A). Similar statistically significant decreases in the cell surface expression levels of GABAAR α2 and γ2 subunits were also evident (supplemental Fig. 1, available at www.jneurosci.org as supplemental material), suggesting that chronic neuronal inactivity leads to the loss of functional heteromeric cell surface GABAARs. In addition TTX treatment also significantly decreased the number of cell surface GABAARs containing β3 subunits in 16 DIV hippocampal neurons
End quote

This is really interesting, with Glutamate antagonists to reduce neuronal activity.

Quote 5
To corroborate our experiments with TTX, we assessed the effects of blocking excitatory synaptic transmission on the cell surface levels of GABAARs with glutamate receptor antagonists, which have been established to reduce neuronal activity in culture (Rao and Craig, 1997; O'Brien et al., 1998). Blockade of glutamate receptors significantly decreased the cell surface expression levels of GABAARs containing β3 subunits to 45.8 ± 13.1% of control (Fig. 1B)
End quote

They used a GABAa antagonist, PTX to increase neuronal activity,

Quote 6
We also examined the effects of chronically increasing neuronal activity on the cell surface expression levels of GABAARs using picrotoxin (PTX) (40 μm), a GABAAR antagonist and has been shown to increase the activity of neuronal cultures (Rao and Craig, 1997; O'Brien et al., 1998). In contrast to the effects of TTX and glutamate receptor antagonists, treatment of neurons with PTX significantly increased both the cell surface (by 55 ± 2.3% of control) and total (by 47 ± 8.5% of control) expression levels of the GABAARs β3 subunit (Fi
End quote

So now the abstract should make sense.....

Quote 7
Abstract
GABA(A) receptors (GABA(A)Rs) are the major mediators of fast synaptic inhibition in the brain. In neurons, these receptors undergo significant rates of endocytosis and exocytosis, processes that regulate both their accumulation at synaptic sites and the efficacy of synaptic inhibition. Here we have evaluated the role that neuronal activity plays in regulating the residence time of GABA(A)Rs on the plasma membrane and their targeting to synapses. Chronic blockade of neuronal activity dramatically increases the level of the GABA(A)R ubiquitination, decreasing their cell surface stability via a mechanism dependent on the activity of the proteasome. Coincident with this loss of cell surface expression levels, TTX treatment reduced both the amplitude and frequency of miniature inhibitory synaptic currents. Conversely, increasing the level of neuronal activity decreases GABA(A)R ubiquitination enhancing their stability on the plasma membrane. Activity-dependent ubiquitination primarily acts to reduce GABA(A)R stability within the endoplasmic reticulum and, thereby, their insertion into the plasma membrane and subsequent accumulation at synaptic sites. Thus, activity-dependent ubiquitination of GABA(A)Rs and their subsequent proteasomal degradation may represent a potent mechanism to regulate the efficacy of synaptic inhibition and may also contribute to homeostatic synaptic plasticity.
End quote

I took lysine 500mg last night after a particularly rough day after I read a post on the board where it helped with sleep. I slept 7 hours straight with no adrenaline surges last night. I had been sleeping 2-3 hours a night with atleast two adrenaline surges a night and none of this. I think by reading this post that lysine works on gaba receptors is that the take away? My benzo brain isn’t really comprehending what the article states.

Hi Jlav

The lysine mentioned in the quote above was referring to lysine residues in the GABAaR itself. As you know, lysine is an amino acid, which is basically a building block of proteins.

In organic chemistry terms you can google it and will find that an amino acid is a simple organic compound containing both a carboxyl (—COOH) and an amino (—NH2) group

GABAaRs consist of (are made up of) proteins. Pers has a good paper on this, but it is not pinned so you will have to search on it. It involves a lot of RNA and DNA stuff.

The study above found that it was the lysine residue in the receptor that made the ubiquitination possible. In very simple terms ubiquitination is the “destruction” or degradation of the GABAaR inside the neuron membrane (this would affect what is called receptor trafficking, which is kind of like how receptors pop in and out of the membrane surface of the neuron, depending upon “need”)

In the study above they replaced the lysine residues with arginine and there was no longer any ubiquitination. It was the ubiquitination that was responsible for the homeostatic response of the neuron to exogenous excitation or lack of (blocking of) excitation. This lines up well with the plasticity that i mentioned in Parts 1 and 2 of the neural circuit dynamics paper. In part 3 we will take high level view of what homeostatic plasticity is (there are many many different types) and how it contrasts to Hebbian plasticity.

Neuroplasticity and Neural circuit dynamics are at the heart of the Benzodiazaphine model, and the answers to the “mysteries” of withdrawal , PWS, and tolerance lie within these areas of neuroscience. It is my belief that therapeutics will come out of research in this area, but our understanding of benzo effects on neural circuit dynamics is just in its infancy. We have a long way to go. A very long way to go.

I’m not sure if the lysine supplement that you took had any affect whatsoever in terms of the articles and quotes above. It may have helped you through more direct mechanisms. I am glad it is helping you.

September 17, 2018

dm123,

I'm sorry I don't remember all the theory.

For a while I was looking at a few things. It's true that not only gaba and glutamate are involved in the process of the action potential, but also neurotransmitters like acetylcholine and serotonin (I could provide links) ?

If that's true, then things like what we eat (serotonin), exercise/stress (acetylcholine) could also affect the action potential ? And thus GABA. You may have put it in different words ...

Hi Liberty, yes. It is true. 5-HTP, DA and ACh and many many other neurotransmitters and neuromodulators have potent effects on the AP of the neurons.

Hence the pillar #5 GPCRs. was added. I don’t think i will be getting to posting much on that pillar, but it is a big part of the benzo model, as are the other 5 pillars.

These neurotranmitters can be thought of as collateral modulators that compensate for the lack of inhibitory signaling present when one enters tolerance. In the same way they would counteract (once again to achieve balance) the overabundance of inhibitory signaling present when benzos are working fine on the GABAaRs. This is one reason (of many) why neurotransmitter levels are altered with drugs like benzos. The emergent circuit behavior must be maintained at all costs, and if benzos are binding to the GABAaRs then the other neurotransmitters will be altered to compensate. If tolerance is excessive these accomodation processes and mechanisms get stretched and if they can no longer accomodate, we get symptoms (circuit distortions, phasic firing relationships break down, etc)

The neurons themselves will respond to overstimulation or understimulation in a homeostatic way, ie it will always strive to maintain balance (not too much AP and not too little AP). The study Jlav pulled from an old ADDENDUM (see above) is one example of this cell level homeostatic response..

Something i never got to is what constitutes too much AP and too little AP. Frequency modulation is how “information” is encoded via APs. The faster the APs fire (more APs per period or unit time), the “more AP” is said to occur, ie the more stimulation generated in the neuron and down its axon. And the more APs per unit time the more neurotransmitters that will be released from that neuron’s presynaptic terminal axons....to the next postsynaptic neuron. So everything is connnected. I’m using terminology really loose to make the point. To make matters even more complex, cable theory must be considered if neural circuit dynamics are being modeled. Cable theory attempts to consider how APs fired down the axon terminal propagate and how much “fade” in the signal occurs down the axon.

THere are intracellular signaling proteins and processes called phosphorylation and de phosphorylation, etc. that control receptor expression from the “inside of the neuron, out”. What goes on inside the membrane is used as feedback by the neuron to control receptor expression on the cell membrane surface, and what goes on at the surface of the neuron (receptors) in turn feeds back to the internals of the neuron.

So you can think of DA, ACh, and 5-HTP as just another set of receptors (called GPCRs which are coupled to ionotropic and other types of receptors) that modulate intrinsic excitability and inhibition inside the neuron.

And remember there are circuit level neuromodulators present that make sure circuit level function is maintained. So you have macro circuit level homeostasis (that affects many synapses....homeostatic plasticity), and micro cellular level homeostasis (like Hebbian plasticity, etc) working together.

I won’t be getting into GPCRs much, but there’s tons of information on them on the internet.

September 18, 2018

DM123

You seem to have a good understanding about all this stuff which I appreciate. I am so happy the lysine helped. I have read not to take anything that works on gaba receptors because this prevents your own GABA receptors to heal or something like that. I don’t really understand all if it. I don’t want to do anything that prevents healing but my thought has always been to try stuff and see how my body reacts. Many things I have not reacted well to but lysine seems to be a good one. I found this article and am not sure what it is trying to say. Would it means lysine is not a good idea in withdrawal?

https://www.ncbi.nlm.nih.gov/m/pubmed/8587651/

September 18, 2018

dm123,

I'm sorry I don't remember all the theory.

For a while I was looking at a few things. It's true that not only gaba and glutamate are involved in the process of the action potential, but also neurotransmitters like acetylcholine and serotonin (I could provide links) ?

If that's true, then things like what we eat (serotonin), exercise/stress (acetylcholine) could also affect the action potential ? And thus GABA. You may have put it in different words ...

Liberty, you are absolutely right. Exercise was/is a major factor for me. I have said it a lot here but I never made the link because I was blaming all the issues only on the taper.

Exercise destabilized my circuits and the next thing you know hyperexcitability sets in and the new level gets established.

"Glutamate receptors form complexes in the brain with auxiliary proteins, which control their activity during fast synaptic transmission through a seemingly bewildering array of effects. Here we devise a way to isolate the activation of complexes using polyamines, which enables us to show that transmembrane AMPA receptor regulatory proteins (TARPs) exert their effects principally on the channel opening reaction. A thermodynamic argument suggests that because TARPs promote channel opening, receptor activation promotes AMPAR-TARP complexes into a superactive state with high open probability."

See the following: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4729862/

Interestingly, in perusing other articles on TARP proteins one article described an evolutionary preserved role for them.

"In vertebrates, at least four TARPs appear to have redundant roles as obligate chaperones for AMPARs, thus greatly complicating analysis of TARP participation in synaptic function. We have overcome this limitation by identifying and mutating the essential set of TARPs in C. elegans (STG-1 and STG-2). In TARP mutants, AMPAR-mediated currents and worm behaviors are selectively disrupted despite apparently normal surface expression and clustering of the receptors. Reconstitution experiments indicate that both STG-1 and STG-2 can functionally substitute for vertebrate TARPs to modify receptor function. Thus, we show that TARPs are obligate auxiliary subunits for AMPARs with a primary, evolutionarily conserved functional role in the modification of current kinetics."

See the following: https://www.ncbi.nlm.nih.gov/pubmed/18817737

So, how do we stop all these TARPs from doing this? Who knows, they have only recently been elucidated. What about modulating post-synaptic P2X Receptors? Good grief, right!

"P2X receptors (P2XRs) are ATP-gated cation channels widely expressed in the brain where they mediate action of extracellular ATP released by neurons or glia. Although purinergic signaling has multiple effects on synaptic transmission and plasticity, P2XR function at brain synapses remains to be established. Here, we show that activation of postsynaptic P2XRs by exogenous ATP or noradrenaline-dependent glial release of endogenous ATP decreases the amplitude of miniature excitatory postsynaptic currents and AMPA-evoked currents in cultured hippocampal neurons. We also observed a P2X-mediated depression of field potentials recorded in CA1 region from brain slices. P2X2Rs trigger dynamin-dependent internalization of AMPA receptors (AMPARs), leading to reduced surface AMPARs in dendrites and at synapses. AMPAR alteration required calcium influx through opened ATP-gated channels and phosphatase or CamKII activities. These findings indicate that postsynaptic P2XRs play a critical role in regulating the surface expression of AMPARs and thereby regulate the synaptic strength."

See the following: https://www.ncbi.nlm.nih.gov/pubmed/25033184

It's a little overwhelming, frankly!!!

-RST

September 18, 2018

The GCPRs ... I was thinking of those as more or less 'fixed' , changable but not quickly.

Another aspect is the effect these have the way they are , causing a certain action under certain acute circumstances such as exercise, food.

'These neurotranmitters can be thought of as collateral modulators that compensate for the lack of inhibitory signaling present when one enters tolerance. In the same way they would counteract (once again to achieve balance) the overabundance of inhibitory signaling present when benzos are working fine on the GABAaRs. This is one reason (of many) why neurotransmitter levels are altered with drugs like benzos. The emergent circuit behavior must be maintained at all costs, and if benzos are binding to the GABAaRs then the other neurotransmitters will be altered to compensate. If tolerance is excessive these accomodation processes and mechanisms get stretched and if they can no longer accomodate, we get symptoms (circuit distortions, phasic firing relationships break down, etc)'

With clonazepam, prior to entering tolerance at least if you're sensitive to that. The antimyoclonic properties of clonazepam are blocked by serotonin antagonists but not gaba antagonists, and the general suggestion is that serotonin is involved. the anti absence properties are suggested to depend on the GABAB component in the thalamic reticular nucleus, where the alpha 3 subunit (??) has immense affinity/effect. Myoclonic seizures, absence seizures.

The effects of chronic stress, affecting the cholinergic and serotonergic system, thus affecting the effects of clonazepam even aside from physical health, and by affecting the cholinergic and serotonergic GCPRs also affecting the GABAA receptors ... sounds like fun !

Comment on that last dose of solifenacin I took ... (antimuscarinic drug). Long half life, aside from anything else, it was sedating for a while. Cross over to diazepam and adding a little solifenacin ... I know crazy ... but it blocks/disrupts signalling from those affected muscles and would relieve some of that 'stress'... It's bad.

September 20, 2018

DM123
You seem to have a good understanding about all this stuff which I appreciate. I am so happy the lysine helped. I have read not to take anything that works on gaba receptors because this prevents your own GABA receptors to heal or something like that. I don’t really understand all if it. I don’t want to do anything that prevents healing but my thought has always been to try stuff and see how my body reacts. Many things I have not reacted well to but lysine seems to be a good one. I found this article and am not sure what it is trying to say. Would it means lysine is not a good idea in withdrawal?

https://www.ncbi.nlm.nih.gov/m/pubmed/8587651/

Yes, if one were to go by that article above alone, it would not be a good idea. It would appear that lysine is modulating the GABAaR. However, i have found many of these “petri” dish style neuron studies to be outdated in terms of explaining PWS, tolerance, etc. because they do not consider the circuit level, nor the homeostatic plasticity responses that occur in real life biological entities.

I’m sure there a tons of other lysine articles like this. 2 were directly linked on pubmed

https://www.ncbi.nlm.nih.gov/m/pubmed/2560434/?i=2&from=/8587651/related

https://www.ncbi.nlm.nih.gov/m/pubmed/1904823/?i=3&from=/8587651/related

These studies are very old and i was unable to pull up the full citation to read what were the methods implored in the lab to arrive at these conclusions. If they were done with live rats or something, there might be some merit to the findings. If they were done with cultured isolated brain slices, its less reliable.

It does appear to affect the Benzodiazaphine binding site on the GABAaR, so it might be potentiating the receptor.

Here is the quote:

“The above results suggest that L-lysine and pentobarbital acted at the same site of the GABA/benzodiazepine receptor complex which was different from the GABA binding site.”

September 20, 2018

The GCPRs ... I was thinking of those as more or less 'fixed' , changable but not quickly.
Another aspect is the effect these have the way they are , causing a certain action under certain acute circumstances such as exercise, food.

'These neurotranmitters can be thought of as collateral modulators that compensate for the lack of inhibitory signaling present when one enters tolerance. In the same way they would counteract (once again to achieve balance) the overabundance of inhibitory signaling present when benzos are working fine on the GABAaRs. This is one reason (of many) why neurotransmitter levels are altered with drugs like benzos. The emergent circuit behavior must be maintained at all costs, and if benzos are binding to the GABAaRs then the other neurotransmitters will be altered to compensate. If tolerance is excessive these accomodation processes and mechanisms get stretched and if they can no longer accomodate, we get symptoms (circuit distortions, phasic firing relationships break down, etc)'

With clonazepam, prior to entering tolerance at least if you're sensitive to that. The antimyoclonic properties of clonazepam are blocked by serotonin antagonists but not gaba antagonists, and the geenral suggestion is that serotonin is involved. the anti absence properties are suggested to depend on the GABAB component in the thalamic reticular nucleus, where the alpha 3 subunit (??) has immense affinity/effect. Myoclonic seizures, absence seizures.

The effects of chronic stress, affecting the cholinergic and serotonergic system, thus affecting the effects of clonazepam even aside from physical health, and by affecting the cholinergic and serotonergic GCPRs also affecting the GABAA receptors ... sounds like fun !

Comment on that last dose of solifenacin I took ... (antimuscarinic drug). Long half life, aside from anything else, it was sedating for a while. Cross over to diazepam and adding a little solifenacin ... I know crazy ... but it blocks/disrupts signalling from those affected muscles and would relieve some of that 'stress'... It's bad.

Yes, the GPCRs play an important part in the model. As i said, i won’t be getting into individual GPCRs, but i will discuss them in the context of the model, and particularly in the context of Neuroplasticity and neural circuit dynamics (ie, in part 3).

So there is homeostatic plasticity which has different temporal aspects from hebbian and other forms of synaptic plastiicty. By temporal, i mean over what time it acts. Most of the mechanisms of homeostatic plasticity act over several (~4) hours to days. So homeostatic plastiicty is a great accommodative mechanism but it is very slow to act. The plastic changes that occur due to homeostatic plasticity are much more profound and often have a greater spatial characteristic than the synaptic plasticity mechanisms. By spatial i mean how much of the neuron, its dendrites, spines and receptors it affects at once. Synaptic scaling is a good example of homeostatic plasticity and i will present it in my own words in part 3.

So when hebbian plasticity changes occur (it is a form of synaptic plasticity), they typically occur quickly on the order of seconds or minutes. Since homeostatic plasticity is so slow to respond, there has to be another mechanism and another form of plasticity that acts to counterbalance the hebbian plasticity changes that occur at the synapse level. There is a form of plastiicty called heterosynaptic plasticity, which occurs at the synapse, is fast acting and is a good candidate for the requirement of a quick homeostatic counter response to hebbian plasticity changes. It just so happens that one form of heterosynaptic plasticity involves GPCRs, in the form of interneurons. These interneurons are typically dopamine and serotonin interneurons (but I’m sure acetylcholine interneurons are involved as well). These interneurons modulate adjacent neuron synapses in a homeostatic fashion in response to synaptic plastiicty changes like hebbian plasticity.

So this is how all these things fit into the benzo model, and how GPCRs as pillar #5 fits into the neural circuit dynamics (Neuroplasticity at the circuit level) system component.

When one takes a benzo, one is either in tolerance or not. If not in tolerance there will be synaptic plastic level changes and if in tolerance there will also be synaptic plastic level changes. Either way, there will be a requirement for a quick acting counterbalancing plastic mechanism, and one (Of many) such mechanism involves these GPCR interneurons that modulate the benzo affected synapses in adjacent neurons.

As I said earlier, this makes sense now, because we know that neurotransmitter levels are all jumbled up when on chronic benzos and regardless of whether or not one is in tolerance. The counterbalancing changes will be in a different direction (tolerance vs. nontolearnce), but there will be changes from baseline of GPCR neurotransmitter levels regardless.

So in this way GPCRs are acting as true neuromodulators. It’s a distinction from their “normal” capacity as excitatory and inhibitory neurotransmitters on the postsynaptic membrane itself. And both of these GPCR functionalities are always at work to maintain homeostasis.

So once clinical research develops better tools in which to study neural circuit dynamics (nanotools), the next step will be to clinically prove all this stuff in real life biological entities, in real time, and with various Benzodiazaphine dosing combos and with various types of benzos. We still need the tools before we can even start to move towards this type of analysis. I am guessing that clinically this is still decades away.

P.S. sorry for typos but typed this out rather quickly. I hope it helps.

September 20, 2018

dm123,

I'm sorry I don't remember all the theory.

For a while I was looking at a few things. It's true that not only gaba and glutamate are involved in the process of the action potential, but also neurotransmitters like acetylcholine and serotonin (I could provide links) ?

If that's true, then things like what we eat (serotonin), exercise/stress (acetylcholine) could also affect the action potential ? And thus GABA. You may have put it in different words ...

Liberty, you are absolutely right. Exercise was/is a major factor for me. I have said it a lot here but I never made the link because I was blaming all the issues only on the taper.

Exercise destabilized my circuits and the next thing you know hyperexcitability sets in and the new level gets established.

"Glutamate receptors form complexes in the brain with auxiliary proteins, which control their activity during fast synaptic transmission through a seemingly bewildering array of effects. Here we devise a way to isolate the activation of complexes using polyamines, which enables us to show that transmembrane AMPA receptor regulatory proteins (TARPs) exert their effects principally on the channel opening reaction. A thermodynamic argument suggests that because TARPs promote channel opening, receptor activation promotes AMPAR-TARP complexes into a superactive state with high open probability."

See the following: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4729862/

Interestingly, in perusing other articles on TARP proteins one article described an evolutionary preserved role for them.

"In vertebrates, at least four TARPs appear to have redundant roles as obligate chaperones for AMPARs, thus greatly complicating analysis of TARP participation in synaptic function. We have overcome this limitation by identifying and mutating the essential set of TARPs in C. elegans (STG-1 and STG-2). In TARP mutants, AMPAR-mediated currents and worm behaviors are selectively disrupted despite apparently normal surface expression and clustering of the receptors. Reconstitution experiments indicate that both STG-1 and STG-2 can functionally substitute for vertebrate TARPs to modify receptor function. Thus, we show that TARPs are obligate auxiliary subunits for AMPARs with a primary, evolutionarily conserved functional role in the modification of current kinetics."

See the following: https://www.ncbi.nlm.nih.gov/pubmed/18817737

So, how do we stop all these TARPs from doing this? Who knows, they have only recently been elucidated. What about modulating post-synaptic P2X Receptors? Good grief, right!

"P2X receptors (P2XRs) are ATP-gated cation channels widely expressed in the brain where they mediate action of extracellular ATP released by neurons or glia. Although purinergic signaling has multiple effects on synaptic transmission and plasticity, P2XR function at brain synapses remains to be established. Here, we show that activation of postsynaptic P2XRs by exogenous ATP or noradrenaline-dependent glial release of endogenous ATP decreases the amplitude of miniature excitatory postsynaptic currents and AMPA-evoked currents in cultured hippocampal neurons. We also observed a P2X-mediated depression of field potentials recorded in CA1 region from brain slices. P2X2Rs trigger dynamin-dependent internalization of AMPA receptors (AMPARs), leading to reduced surface AMPARs in dendrites and at synapses. AMPAR alteration required calcium influx through opened ATP-gated channels and phosphatase or CamKII activities. These findings indicate that postsynaptic P2XRs play a critical role in regulating the surface expression of AMPARs and thereby regulate the synaptic strength."

See the following: https://www.ncbi.nlm.nih.gov/pubmed/25033184

It's a little overwhelming, frankly!!!

-RST

Yikes !!!, that is some heavy stuff. I will need to read it over.

Initially i thought you were referring to TRPs or transient receptor potential channels, which interestingly enough are heavily associated with neuropathic pain (and i mentioned them a bit during the NO/ONOO- discussion0. However, TARPs are a totally different thing.

Here’s a bit on TRPs which are not TARPs. The research in this area shows some promise for neuropathic pain, and there are lots of articles on it (full citations)

https://en.m.wikipedia.org/wiki/Transient_receptor_potential_channel

September 20, 2018

I suppose there is no specific theory about getting the circuits 'back in order' ? I feel like it's a dumb qustion.

September 20, 2018

I suppose there is no specific theory about getting the circuits 'back in order' ? I feel like it's a dumb qustion.

Not a dumb question at all, Liberty. Rather, the BILLION dollar question!!!

-RST

September 21, 2018

DM123
You seem to have a good understanding about all this stuff which I appreciate. I am so happy the lysine helped. I have read not to take anything that works on gaba receptors because this prevents your own GABA receptors to heal or something like that. I don’t really understand all if it. I don’t want to do anything that prevents healing but my thought has always been to try stuff and see how my body reacts. Many things I have not reacted well to but lysine seems to be a good one. I found this article and am not sure what it is trying to say. Would it means lysine is not a good idea in withdrawal?

https://www.ncbi.nlm.nih.gov/m/pubmed/8587651/

Yes, if one were to go by that article above alone, it would not be a good idea. It would appear that lysine is modulating the GABAaR. However, i have found many of these “petri” dish style neuron studies to be outdated in terms of explaining PWS, tolerance, etc. because they do not consider the circuit level, nor the homeostatic plasticity responses that occur in real life biological entities.

I’m sure there a tons of other lysine articles like this. 2 were directly linked on pubmed

https://www.ncbi.nlm.nih.gov/m/pubmed/2560434/?i=2&from=/8587651/related

https://www.ncbi.nlm.nih.gov/m/pubmed/1904823/?i=3&from=/8587651/related

These studies are very old and i was unable to pull up the full citation to read what were the methods implored in the lab to arrive at these conclusions. If they were done with live rats or something, there might be some merit to the findings. If they were done with cultured isolated brain slices, its less reliable.

It does appear to affect the Benzodiazaphine binding site on the GABAaR, so it might be potentiating the receptor.
Here is the quote:
“The above results suggest that L-lysine and pentobarbital acted at the same site of the GABA/benzodiazepine receptor complex which was different from the GABA binding site.”

Thank you for your input. It seem anything that May help is a no go.

September 21, 2018

DM123
You seem to have a good understanding about all this stuff which I appreciate. I am so happy the lysine helped. I have read not to take anything that works on gaba receptors because this prevents your own GABA receptors to heal or something like that. I don’t really understand all if it. I don’t want to do anything that prevents healing but my thought has always been to try stuff and see how my body reacts. Many things I have not reacted well to but lysine seems to be a good one. I found this article and am not sure what it is trying to say. Would it means lysine is not a good idea in withdrawal?

https://www.ncbi.nlm.nih.gov/m/pubmed/8587651/

Yes, if one were to go by that article above alone, it would not be a good idea. It would appear that lysine is modulating the GABAaR. However, i have found many of these “petri” dish style neuron studies to be outdated in terms of explaining PWS, tolerance, etc. because they do not consider the circuit level, nor the homeostatic plasticity responses that occur in real life biological entities.

I’m sure there a tons of other lysine articles like this. 2 were directly linked on pubmed

https://www.ncbi.nlm.nih.gov/m/pubmed/2560434/?i=2&from=/8587651/related

https://www.ncbi.nlm.nih.gov/m/pubmed/1904823/?i=3&from=/8587651/related

These studies are very old and i was unable to pull up the full citation to read what were the methods implored in the lab to arrive at these conclusions. If they were done with live rats or something, there might be some merit to the findings. If they were done with cultured isolated brain slices, its less reliable.

It does appear to affect the Benzodiazaphine binding site on the GABAaR, so it might be potentiating the receptor.
Here is the quote:
“The above results suggest that L-lysine and pentobarbital acted at the same site of the GABA/benzodiazepine receptor complex which was different from the GABA binding site.”

Thank you for your input. It seem anything that May help is a no go.

So far it's not proving to be easy to find something that is directly helpful with the problem. At best we have some discussion around support systems and various supplements which have anecdotally been helpful for some and otherwise for the rest of us. I have come to regard the CNS as a large group of jugglers assembled in a circle who collectively juggle all the balls in an intricately synchronized pattern that includes juggling several balls individually as well as exchanging balls through the air with neighbors to either side as well as with team members assembled in different positions around the circle. Benzodiazepines are like a new juggler that joins the group with 1 cool trick but eventually grows bored and when he suddenly leaves the group the beautiful synchronicity dissolves in a catastrophic crash. Sometimes you can cajole him to rejoin the group but he's still bored and doesn't want to be there so his effort is half-hearted and the synchronicity only kind of returns. Eventually, the other jugglers spend so much time dealing with the recalcitrant benzo-juggler that they start dropping more balls and suddenly they can only keep a few critical balls in the air while the rest are ignored. Now, the team can only but exist. The team would prefer he leaves altogether but know he will likely steal some of the balls when he goes this time and they risk never getting them back!

It's a silly analogy of course but it's what I see in my mind when I'm visualizing a progressive return of effiecieny to my CNS jugglers.

-RST

Can anyone make sense of this to explain to a layperson?

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