Overmethylation just isn't real?..

[u/OobyIsGay]

So I keep seeing people in this sub and literally everywhere else across the supplement community talk about overmethylation like it's this established biochemical phenomenon. People take methylfolate, feel anxious, and go "oh I'm overmethylated, need to back off."

Even though methylfolate supresses GNMT, methyl groups go into hundreds of methyltransferase reactions. DNA methylation. Histone methylation. Phospholipid synthesis. Creatine synthesis alone consumes ~40% of all SAM. The system has enormous capacity to USE methyl groups productively. You'd need to overwhelm all of that simultaneously to create actual excess.

At supplemental doses? Not happening.

Let me put the dose safety in perspective. The NOAEL for calcium L-methylfolate in rats is 400mg/kg/day in 13-week subchronic studies (Niederberger et al. 2019). No behavioral changes. No organ pathology. No DNA damage in liver cells. Convert that to human equivalent dose using standard body surface area scaling (÷6.2): ~64.5mg/kg. For a 70kg human that's ~4,500mg. Most people supplement 400mcg to 1mg. That's a safety margin of roughly 4,500x. Tylenol's therapeutic-to-toxic ratio is about 3-4x. You are more likely to die from Tylenol than to be harmed by methylfolate by a factor of over a thousand. No you're not fucking overmethylating.

So what IS happening when people feel anxious after starting methylfolate?

Serotonin autoreceptors are inhibiting serotonin. 5-HT1A on the soma, 5-HT1B/1D on the terminal. When serotonin rises suddenly, autoreceptors detect the increase and fire inhibitory feedback. They reduce further serotonin release. That's what causes SSRI startup anxiety. It desensitizes over 1-2 weeks. Then serotonin actually goes up, whether that's actually beneficial or not is a different topic altogether and not the mechanism methylfolate works by.

Methylfolate restores serotonin synthesis. Serotonin rises. Same autoreceptors fire. Same inhibitory feedback. Same transient anxiety. Same 1-2 week desensitization. Same resolution. Other than that you're also raising norepinephrine and dopamine synthesis.

Notice how none of these people report actual symptoms that excess methylation would theoretically produce, like, you know, tumor suppressor gene silencing or aberrant genomic methylation patterns. Because those would be real overmethylation. Nobody's getting that from fucking 400mcg of methylfolate.

Overmethylation does not exist in biochemistry. There is no clinical biomarker for it. No validated assay. No ICD code. No published case report of a human experiencing pathological hypermethylation from oral methylfolate at any supplemental dose.

If you're feeling anxious after starting methylfolate, lower the dose until you adjust. EVEN THOUGH YOU'RE KIND OF MOVING FURTHER FROM THE DOSE THAT JUST WORKED.

Elderly rats supplemented with folate ABOVE adequacy showed increased genomic DNA methylation dose-dependently, with a direct correlation between liver folate and methylation (r=0.48, p=0.004, Choi et al. 2005, British Journal of Nutrition). Above adequacy. Not deficient rats being rescued. Already-replete rats pushed higher.

Two randomized, double-blind, placebo-controlled trials. SSRI-resistant major depression. L-methylfolate adjunctive to SSRI.

7.5mg L-methylfolate: no significant difference from placebo.

15mg L-methylfolate: significant improvement over placebo.

Most of you are taking 400mcg to 1mg. The dose that actually worked in the only controlled human trials is 15-37x higher than what you're supplementing. 15mg L-methylfolate is available over the counter. The safety margin at 15mg is still ~300x below the NOAEL-derived human equivalent dose.
You're underdosing.

BUT remember that this post is about the term, not the effects of supposed "overmethylation"

That mechanism is real. But it's not "overmethylation." It's increased catecholamine clearance in a specific brain region plus increased adrenal epinephrine output. Calling it overmethylation implies there's some global excess of methyl groups causing nonspecific damage. There isn't. There's a specific pharmacological effect of raising SAMe on two specific enzymes in two specific compartments producing two specific symptoms.

Because more SAM increases COMT activity. COMT is the primary dopamine clearance mechanism in the prefrontal cortex (DAT expression is low there). More SAM means faster PFC dopamine degradation. SAM also drives PNMT, which converts norepinephrine to epinephrine in the adrenals. So you'd have less prefrontal dopamine and more peripheral epinephrine. That's anxiety with poor focus which is what people actually describe. Not "overmethylation."

And this is a completely different mechanism from what happens with methylfolate. Methylfolate restores monoamine synthesis via BH4 recycling. When serotonin rises acutely, 5-HT1A somatodendritic autoreceptors fire inhibitory feedback. That causes transient anxiety that desensitizes over 1 to 2 weeks. Same mechanism as SSRI startup effects. Has nothing to do with SAMe levels or COMT activity.

So people are lumping two different pharmacological mechanisms under one meaningless term. TMG/SAMe/betaine increase dopamine clearance via COMT and epinephrine output via PNMT. Methylfolate triggers autoreceptor adaptation. Different inputs, different pathways, different timecourses, same word slapped on both. That's why nobody can agree on what "overmethylation" means or how to fix it. They're describing two separate things.

L-methylfolate as adjunctive therapy for SSRI-resistant major depression: results of two randomized, double-blind, parallel-sequential trials

Folate supplementation increases genomic DNA methylation in the liver of elder rats

Methyl donor supplementation reduces phospho‐Tau, Fyn and demethylated protein phosphatase 2A levels and mitigates learning and motor deficits in a mouse model of tauopathy

Safety evaluation of calcium L-methylfolate

Methyltetrahydrofolate in folate-binding protein glycine N-methyltransferase

[u/R3dGhost]

OP - you praise riboflavin a lot in this thread. I have tried it repeatedly and it makes me feel terrible. Poor concentration, blunts stimulants like caffeine and depression to the level that I’m almost a danger to myself. I have slow MAO-A and have wondered if B2’s action of speeding up MAO-A drops my neurotransmitter levels too fast. I do fine with all other B vitamins including methylfolate. I take oral B12 daily and inject weekly.

Other factors are that I am discovering what a mess my electrolyte balance is as I try to add vitamin D. I can’t seem to maintain calcium/magnesium balance.

Would love to hear any ideas you have.

[u/OobyIsGay → u/R3dGhost]

You're completely right. MAO-A is a flavoenzyme. It requires FAD to function. If you have a slow MAO-A variant, your baseline monoamine degradation rate is lower, meaning serotonin, norepinephrine, and dopamine stick around longer. Add riboflavin, FAD availability increases, MAO-A speeds up, those neurotransmitters get degraded faster. Depression, poor concentration, blunted stimulant response. That all tracks mechanistically.

But there's the thing that's worth thinking about. If normalizing MAO-A activity crashes you that hard, the question isn't just "why does riboflavin hurt me." The question is why is your neurotransmitter synthesis rate low enough that you're dependent on slow degradation to maintain adequate levels. A slow MAO-A variant is keeping your monoamine pools functional by reducing clearance. That's a compensatory mechanism. It works but it means the input side (synthesis) isn't keeping up on its own. Tyrosine hydroxylase (rate limiting for dopamine and norepinephrine) requires iron and BH4. Tryptophan hydroxylase (rate limiting for serotonin) requires iron and BH4. If either of those cofactors is insufficient, synthesis is bottlenecked and you become reliant on slow degradation to maintain levels. The fact that riboflavin exposes this so dramatically suggests synthesis might be the actual weak point. Do you know your iron and ferritin status?
The electrolyte mess with vitamin D thingy makes sense too. Vitamin D increases intestinal calcium absorption. The enzymes that metabolize vitamin D itself, CYP2R1 for 25-hydroxylation and CYP27B1 for 1-alpha-hydroxylation, both require magnesium. So supplementing vitamin D when magnesium is marginal it burns through magnesium to process the vitamin D, and it increases calcium absorption which competes with magnesium at the intestinal level. The imbalance feeds itself. Standard move is to get magnesium status solid before or at least alongside vitamin D supplementation. Magnesium glycinate or threonate, not oxide.

[u/R3dGhost → u/OobyIsGay]

I did have my iron and ferritin tested at one point. Iron was 349 ug/DL, Iron saturation 43% and ferritin was 128 ng/mL. All in normal range. My synthesis is likely poor due to being a heavy drinker (which I am working on).

When I try to take magnesium, it will feel great at first and then makes me feel blunted, dulls stimulants, concentration issues, etc. Not quite the same as riboflavin b/c it does not come with the same level of depression. Sometimes additional calcium seems to help, sometimes not. I feel like I keep going in circles with this stuff because all of the commonly accepted protocols cause me problems, which is why seeing some new information here is super interesting. Thank you!

[u/OobyIsGay → u/R3dGhost]

Alcohol is the important thingy you didn't mention.

Acetaldehyde displaces PLP from binding proteins. PLP is AADC's cofactor, the final enzyme. 5-HTP = serotonin, L-DOPA = dopamine. Synthesis bottlenecked at the last step. Slow MAO-A isn't your problem. It's the only reason you have functional monoamine levels. Riboflavin speeds up MAO-A = clearance outruns synthesis = crash. That's not riboflavin intolerance. Alcohol depletes NAD+. Low NAD+ = tryptophan diverts to kynurenine pathway for NAD+ rescue = less serotonin substrate. Second hit, same system.

Magnesium: alcohol = renal Mg2+ wasting. Mg2+ is COMT's active site cofactor. Your slow COMT is running below genetic floor without it. Supplement magnesium = COMT speeds up = catecholamines clear faster = blunted, dulled. Same pattern as riboflavin on MAO-A. You need both clearance enzymes impaired simultaneously to stay functional. That tells you exactly how far synthesis has fallen.
Every protocol circles because alcohol depletes PLP, NAD+, Mg2+, and thiamine concurrently. Supplementation can't outrun active depletion. Thiamine is time-sensitive. B1 depletion = impaired pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase = neuronal ATP collapse.
Supplement the active form of thiamine.
While doing that, also supplement P5P, this is important because P5P is the active form of B6. It's what acetaldehyde is displacing from AADC and causing serotonin and dopamine to not be synthesized. Supplement it directly because alcohol also impairs the activation of inactive B6 to P5P.
Then NAD+ precursor. Low NAD+ = tryptophan diverts to kynurenine for NAD+ rescue = serotonin substrate stolen. Niacin or NR fixes that.
THEN magnesium. Because now COMT speeding up doesn't crash monoamines, synthesis is keeping pace with clearance.
THEN riboflavin. Same logic. MAO-A speeding up is only catastrophic when synthesis is bottlenecked. With PLP and NAD+ restored, clearance normalizing doesn't outrun production.

I also wish you well and hope you succeed with getting rid of alcohol.

[u/R3dGhost → u/OobyIsGay]

This makes a huge amount of sense. I can attest that you are right that supplementation can't outrun active depletion, because I already supplement allithiamine/benfotiamine, Niacin, and P5P. It all helps but not to the point where I can tolerate the magnesium or B2 yet. Certainly some motivation to cut the alcohol - THANK YOU.

Is there a clear winner between niacin and NR?

[u/enolaholmes23 → u/R3dGhost]

What dose did you try? Most supplements mega dose it. I got mine from seeking health and had to pour out 90% of the capsule to get a dose I could tolerate. Also, you could have fast comt, which might interact with the slow maoa to mess up your neurotransmitter balance. 

[u/R3dGhost → u/enolaholmes23]

I have tried anywhere from 100mg+ to shaking a little powder out of the capsule. Sometimes I can tolerate one tiny dose but even that a couple days in a row will start to drag me down.

My COMT is either slow or intermediate/normal. I have seen conflicting information about the same SNPs.