Thanx for the links, benzyme. That was kind of you.

Arrivederci

Benzyme posted the KEGG pathway of tryptophan metabolism, it is pretty well known, as im sure you are well aware. Here's another link to serotonin biosynthesis, and as you see all the enzymes necessary for it are found in the human brain (as opposed to DMT), such as tryptophane hydroxylase and aromatic l-amino acid decarboxylase (AADC) .




Is there any sign anywhere in nature (and specially in man) of methylation that is not done by the known methyltransferases and therefore would point to a significant hole in the endogenous trace amine production pathway?



Im not aware of other experiments as detailed as that one regarding INMT expression.

Did you read the INMT research article? Is there anything in the method you think is a possible weakspot?

I guess we'll just have to disagree on this one then, while you think that 'the fact experiments can be wrong' is good evidence, I personally feel that before one points out a significant flaw in the experiment method and/or repeats it and shows otherwise, there is no reason to a priori consider it wrong and think that this is evidence for some alternative explanation.



Maybe you're right, but considering these pathways in mammals are AFAIK explained by methyltransferases leaves you again with a sticky problem. You would probably think about 'mistery uncharacterized enzymes', which I would love to get some more data about this if you can link me up. So far though it doesnt seem to me like enough evidence to call it 'very likely'. And damn you for not giving me the grant for my dmt-in-the-ear study!





Oh do notice I started answering from when lodi used history channel as source of evidence and said it is 'real' for sure. Here in the Nexus (and anywhere in the scientific community) history channel wont be considered good reference, and to say anything with certainty such as dmt production here will need to be accompanied by good sources.

Then of course the discussion continued with you. You are right you didnt use unreliable sources and I appreciate the discussion, though I will disagree about what evidence points to, before the biggest hole is really explained. So my request for reliable sources and not spread misinformation was a more general one. Considering your generally reasonable way of putting yourself, that request isnt really directed at you.

What I would ask from you though is that if you ever do say around the Nexus (maybe elsewhere too? though thats up to you of course) that it is very likely DMT is produced in the brain, that you make it clear that a fundamental enzyme is still missing from that equation and hence its still very debatable what evidence is pointing to.



No worries I didnt feel anything negative about the bet, just really wanted to express that my skepticism and prudent orientation isnt implying either way of the discussion, to me it would all be interesting.

But if it makes you happy I bet you some potatos and onions, that would save me some walk to the market



Nope I agree there is no conclusive evidence it is a misconception but its definitely only speculation at this point, so still better that we are clear about it



Thanks, agreed the site is awesome and look forward to future dialogues too. See ya

Endless thanks again for the linking article. I've read it and I stand by my belief for a couple of reasons.

First off I think these guy's did some good work, but I'd still like to see it repeated in another lab....hear me out.

1) It's just nice to have independent results that match.

2) I agree with them that based on the Ki of INMT on human tryptamine (2 micro molar) that it's role in humans may very well be something other than N-Methylating Tryptamines. In the rat it was (~0.2 micro molar) in human it (~2 micor molar) or an order of magnitude difference. To be sure there are some human protein/ligand complexes that have micro molar affinities, but they are the exception and not the rule in my experience. So seeing that they are finding N-methylated tryptamines I say it's still very plausible that another route could be found. I also say it's plausibly that it's just a trace byproduct of INMT that eventually gets into the brain. My gut still thinks it's produced in the brain since it's found there. Since it's a trace amine, perhaps the neurons that produce it are also found in very trace quantities? I'm not ready to through out INMT as the explanation yet either because of....

3) They did find INMT it in the spinal cords which means methylation hardware is found in the CNS... I'd like to see where it was isolated from the spinal chord. What kind of cells? Perhaps it would give some clues about MORE specific brain areas to search in.
"It was of interest that, even though this mRNA was not expressed in either total brain preparations or tissue obtained from specific sublocalizations within the brain, it was well expressed in the spinal cord. The possible functional implications of that observation should be explored in the future."

4) If you look at Fig 2....I do believe I see trace spots for INMT in a couple of brain sections. Notice the faint line around ~2.5 KB's. It's most likely noise, but notice that there is total absence of noise on the whole brain run. Also the positve control using B-actin appears to show something for the corpus callosum that is above the noise level. I'm going to openly admit I'm outside my area a little bit here.... I'm going to talk to a couple of biologists at work today who I'm sure will have a much better handle on this as they run western blots quite frequently.

6) The pineal gland is completely absent from their study...yes it's just outside the BBB..but still part of the brain none-the-less. I suppose we could debate that...but in any event I'd sure like to see what pops up with the pineal used alone. The pineal obviously has the methylating enzymes at it donates a methyl group to the 5-OH of serotonin in melatonin production. That suggests to me that s-Adenosyl transferase is involved and quite possibly INMT as well I mean how else to you account for melatonin being methylated if there aren't any methylation pathways? I'm going to again claim a little ignorance, because I suppose that O-Methylation is competely different from N-mehtylaltion...this will give me something to read up on!

So I guess what I'm saying is that no I'm not willing to toss the baby out with the bath water over one study.
The evidence as I see it is:

1) DMT has been found in nano molar concentrations in the brain.
2) The pineal gland has Methylation enzymes as evidenced by the formation of melatonin.
3) DMT production has been verified in other mammals.
4) I doubt even trace amounts can make it to the brain and survive MAO metabolism.
5) One study showed that INMT wasn't found in the brain.
6) The same study found it in the CNS.

Based on the above I say there are 4 points #1,#2, #3, #6 that are hinting at DMT being produced in the brain, and one #5 showing it's not...and #6 is my speculation about the MAO system being to robust to find even trace amounts of DMT in a brain full of MAO after more than a few minutes...

We don't have to agree about which direction the evidence points...it doesn't really matter until someone finds out for sure. I'm also not shooting down the study you listed. I'm just siding with caution because to me the simplest answer is still that it will be found to be produced in at least a small degree in the brain.

I also thing that anyone that frequents the nexus more than a couple of times becomes aware of the fact that the pineal is just a theory of Dr. Strassmans. Yes a lot of web sites are touting it as a fact and they are wrong...it's not a fact, but...well you already know what I'm going to say!

Anyway I will post back if the biologists at work have any additional insights. If not I think it's time to let this thread die down. Your points about not spreading misconceptions and false information are valid. DMT-Nexus has been one of the better sources for information out there. I'd like to keep it that way as well since it's one of the only places on the internet I call home!

<---------- Newbie here who has never tried a psychedelic... ever, yet.

I enjoyed this conversation, and want to throw a few thoughts into it that have nothing to do with the science. (BTW, for those who aren't biochemist types in here like me, for the most part, a good starting point is The Biology of Belief by Bruce Lipton... novice terms for cellular makeup/behavior imho.)

It is obviously a good thing to study the biology behind any physical process we are curious about. It is easy to understand the appeal to many, especially when very personally involved in the attempt to validate (for various reasons) a currently illegal and stigmatized substance/practice. The human propensity to label everything in any environment is without question and obviously deeply ingrained in our conditioning. It may come from a primal psychological need to understand the mysterious, thereby making it less fearful (not suggesting anyone here is doing that, just pointing out what we all have in common to some degree). While there is obviously incredible usefulness in this trait, I wonder if we extend it into realms of experience that cannot possibly be begun to be explained by science when our current scientific limitations are inadequate at best, and of an entirely different "dimension" at worst.

The potential of the human brain, that is, the potential of conscious awareness, is still a vast mystery. That is an infinite understatement. A quick analogy: To fully understand everything of the biochemical process of any psychedelic, in relation to the formless experiences and truths the process can point to, would be to fully understand the marital status of a leaf on a rose bush. Ultimately, the questions are unanswerable. It's the difference between knowledge (which ultimately is limited by the phenomena of human perception) and experience. In other words, maybe a mental concept of attained knowledge has nothing to do with the implications of an experience that far transcend the relative scale of one branch of science. From what I have gathered from what has been posted on this forum, is this not the case?

I guess the point is this: let's say that tomorrow, irrefutable proof is published that states DMT (or even just practiced meditation) is caused by __________, and all biochemical processes are mapped, tracked, explained, etc... Will the "knowledge" of this really add to the ultimate comprehension of these experiences, and their implications?

Certainly the intent here is not to disvalidate this particular pursuit of knowledge, but only to put it in perspective while enjoying the attempt.

It sure would be nice to know

Sorry to keep this alive,but I just spoke to our head of biology and would like to share his thoughts on this....I trust him a good bit more than myself when it comes to biological assays.

Looking at figure 2, Part C, he said that the spots we are seeing for the various brain regions are in all likely hood just noise. He walked me through the entire experiment protocol and I agree with him fully.

He did however make the exact same observation that I did about the spinal cords and was pretty intrigued by it.

I walked him through the various evidence for and against it and he can see it both way's (as can I), but ultimately he seems to think that the simplest answer is that it will be produced there.... So that's at least 2 scientists that share that view. one biologist and one chemist.

One further point that he was able to shed light on that I wasn't even considering is the mRNA expression. Note they are measuring the mRNA and not DMT in this experiment. It could very well be that the mRNA's for INMT isn't expressed in the brain at all times. Just like the mRNA's for the enzymes necessary for conversion of melatonin are likely to be absent or very reduced during the noon sun light. I don't have links here, but his argument was pretty compelling that mRNA's are used almost as fast as they are made and are then metabolized to keep the expression from becoming infinite. Makes sense, but I haven't done any leg work here.

So if go back to Rick's original hypothesis that DMT is produced in the pineal gland during times of stress...I'd have to say that this paper does not address that theory in a very good manner for a couple of reasons:

1) They don't address the pineal gland in this research
2) They aren't using a brain that is stressed.

I say Rick's theory is still a pretty reasonable theory in light of the evidence I've looked at thus far. Certainly it's not proof, but it's certainly not proven otherwise either....or at least I have not come across said proof.

Questions I'd like to answer:

1) Does the cerebral spinal fluid have direct access across the BBB? There results clearly show INMT in the spinal cord. It could very well be coming from there....and potentially not have to cross the BBB and be subject to higher MAO concentrations. I'm intrigued about this results from a metaphysical point of view as well, but I'm not going down that path in a science discussion... That thread will come later when I've done more fleshing out of the idea!

2) Does the pineal gland produce INMT or other N-Methylating enzymes and if it so does it do it at different times of the day? We'd obviously expect it at night since melatonin is produced then. But is it found during the day as well...and if so is the concentration greatly reduced?


I still keep my friendly,no wager, bet alive. I say within 2 years we have the proof. Endless I'm not coming to you for potatoes, but if I'm ever on your side of the pond I'd meet up for a beer for sure!

I have no shame in being wrong. I'm just judging the theory as it I see it. the beauty of science is that it won't care what we think. The data will tell the final story.

Peace

Keep an open mind and a positive attitude, and soak up good info and you are on the right path before you have even tried a psych. This is the main goal, to take away the nonsense and stick with what we know to be proven but open to new information. An open mind, a flexible attitude like that is important, but to make false claims only clouds these waters. You are correct when you state we are not trying to invalidate the pursuit of knowlege.

This nexus is full of intellectuals, chemists, mystics, etc, and everyone is entitled to their own opinion. But to claim something as fact with nothing to back it up seems to go against the pursuit of true knowledge.

Also nothing wrong with debating in here, this topic really puts it into perspective.

Good work there joedirt, I appreciate the investigative effort. If only more people would be like this... Some interesting observations and food for thought! I'll research some more on this and come back to the discussion.

I must agree that there is no evidence that shows dmt is definitely not made in the pineal and that more specific research could help elucidate our question. But again I see no reason to also jump the other way and say that this is evidence that dmt is made in pineal, but I respect your opinion if from your position you see the likelihood in this direction of biosynthetic pathway. Personally I choose to abstain at this moment from believing one or the other and will just wait for new information to come up. I agree that the beauty is that truth comes out at the end and Im also happy with whichever truth that is

The meet up sounds fine by me, I would preffer some juice though

Very interesting! Thanks joedirt

HIOMT

not INMT.
the former is well referenced, and the latter isn't magically expressed in the pineal when stressed. no mRNA activation if the mRNA transcripts aren't present.

again, separate pathways in tryptophan metabolism. show evidence otherwise, then this may be refuted.

in the pathway, you'll notice two separate pathways where INMT is expressed... the first path is N-methylation of tryptamine, and the second is N-Methylation of serotonin. activation of the first path inactivates the second.
the second, analogous to the first, likely leads to bufotenine.

Ok I've done a little more research, no it's not complete, but I want to offer a couple of more thoughts to this thread.

I believe most people are interested in whether DMT is produced in the brain because that would make it an endogenous neurotransmitter. Today I did enough research to convince myself that it is indeed an endogenous neurotransmitter whether or not it's made in the brain because the proper N-Methylations enzyme INMT is found in the spinal chord which is surrounded by the cerebral spinal fluid which flows uninhibited throughout the CNS. At this point the main thing needed to lock this down as fact is proof that the spine produces DMT...being that the spine has INMT and tryptamines like serotonin floating around I'd be pretty well amazed if there weren't N-methyl tryptamines there as well.

The worst source I'll cite...if it's really important I'll dig out my Anatomy and Physiology book...
http://users.rcn.com/jki...S.html#BloodBrainBarrier

But these wiki's cover it in more words...
http://en.wikipedia.org/wiki/Cerebrospinal_fluid
http://en.wikipedia.org/wiki/Blood-brain_barrier

Combined with thae fact that we now have trace amine receptors in the brain I'd say that for me at least I'm beginning to believe that DMT is an endogenous neurotransmitter even if it's not produced in the brain directly.



Now back to whether or not DMT is produced in the brain...

There are potentially other N-methylation enzymes that are found in the brain which "could" possibly catalyze trace amounts of tryptamines. The first one I'd suspect would be:

phenylethanolamine N-methyltransferase (PNMT) which is found in the adrenal medulla and is known to converts Norepinephrine to Epinephrine.

http://en.wikipedia.org/...mine_N-methyltransferase


These are phenethylamine and not tryptamine enzyemes, but being that both add a methyl group to a primary amine makes it seem plausible. I have not as of yet looked at the literature to see if someone else has checked to see if tryptamine undergoes an N-methylation in the presence of this enzyme.. More work I suppose. I'll try and dig that up later when I have more time.

Another reason PNMT seems plausible to me is that many of the receptors in the brain bind several classes of neurotransmitters. For instance serotonin has some affinity for dopamine receptors and vice verso. Of course the non specific affinities are much higher. But in the case of trace amounts of DMT it seems plausible that a small amount of tryptamine or serotonin could be N-Methylated via this route. If it happened to serotonin we should be able to find some 5-methoxy DMT floating around as well because we know for sure that the pineal gland has O-methylation enzymes. Guess we will have to wait on LSU's endogenous trace amine assay to come online...I wonder what the status of that is?

There is also histamine N-methyl transferase which is found in most body tissues but is not present in fluid. Histamine is also a primary amine, but I don't as of yet know it it's found in the brain.
http://en.wikipedia.org/...mine_N-methyltransferase


So from here on out when someone says is DMT made in the brain my answer is going to be, "We don't know for sure, but the proper enzymes are found in the Spine which is surrounded by the cerebral spinal fluid which provides direct access to the brain...."

That's my story and I'm stick'in to it...well until I see other evidence to change my mind!

I agree about INMT vs HIOMT I just need to look into it a bit more...see my post before this one. I'm also very well aware of the tryptaphan metablism and conversion in the body. I'm also well aware that most of the pathway's people show don't include all the side products and trace pathway's that are also present. Since DMT is a trace amine it's plausible to argue for a minor route in the body.

Can you explain this part to me. "no mRNA activation if the mRNA transcripts aren't present. "

DNA is present in all cells... What is the mRNA trnscript that isn't present...and can you provide a link for this...sorry my biology is a little rough.

all i know is that in autopsy's of schizophrenics they did find higher concentrations of dmt im not sure if it was in the brain or somewhere else in the body tho

the so-called 'central dogma' of biochemistry is DNA --> mRNA --> protein.
INMT is obviously a protein; therefore, the transcript must be present in the cell for expression.
while it's true that DNA is present in every cell, less than 2% of human DNA encodes for protein synthesis.

you can find this in any biochemistry textbook.

yeah I know...I was being a smart ass. I was just wondering if you were going to contribute references or links to back up your statement that the mRNA coFactors aren't ever present. I mean at this point it's pretty obvious that we are just speculating in the realm of what's scientifically possible. I have no skin in the game and if you can show references that prove that I'd be happy to rule it out as a possibility!

Interesting thread.

Are any other endogenous substrates of INMT known?

Concerning methylation by other methylating enzymes. Its possible and it can at least be measured simply in vitro to see if the reactions can occur at all. Just can't say for sure until someone does some assays.


I found a paper that demonstrates N-methylation of tryptamine to n-methyltryptamine occurs in human brain tissue removed during surgery:

Psychomimetic N-methylated Tryptamines: Formation in Brain in vitro and in vivo. Juan M Saavedra and Julius Axelrod (haha his name is axelrod )
Science Volume 175.

Its an old ass paper. Someone should repeat (or has it been repeated?) using modern chromatographic analysis and gene expression analysis.

yea, i posted them earlier, smartass
gave you food for thought, didn't it?
I've actually posted several references on the topic, scattered throughout the nexus, i'm just too lazy to dredge them all; it's a topic that's been kicked around and beaten to death.
nevertheless, it's always an interesting puzzle shrouded in mystery, and the current model
doesn't show any significant production in the brain; not that it won't be shown otherwise, but i wouldn't hold my breath. it's pretty obvious that the chances are greater for DMT and bufotenine production in the peripheral tissues, being that 80-90% of serotonin is produced in the gut.



unlike AADC, I think its exclusive to endogenous tryptamines.
the binding pocket is probably structured to accept that indole moiety, excluding catecholamines, which lack the pyrrole



I think that study was scrutinized by Thompson et al. (1999), and this reviewed by Presti et al. (2004).

Big ups to you all for bringing up this subject ,, it's a shame that such an interesting thing like this don't get research funds

I couldn't follow most of the discussion going on here,, but I am secretly hoping that there is truth to the pineal and DMT relation
because pineal is where the third eye is,, and I have strong personal faith in ancient chakra knowledge

what if scientific studies showed evidence that DMT is produced in epithelial cells near the sphincter? would that disappoint you?

It's also produced in the spine which is present in the CNS and has direct access to the brain via the cerebral spinal fluid.

Your point is more than valid....ie...it is produced in the body, it can cross the BBB...and there is no evidence yet that it's produced in the brain.


That facts are that DMT is produced in the CNS, found in the brain, and that the INMT (N methylation enzymes) is not found in full brain scans, but the studies that have been performed ( to my knowledge ) do not cover the pineal gland....it is after all outside the Blood Brain Barrier.



I love mystery! It challenge's man to debate!

Sorry I need to amend my statement there. There is no proof the DMT is produced in the spine. However INMT ( The n methylation enzymes) are present...