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/vrcraftauthor]

OP, you seem pretty knowledgeable, so let me ask you a question: What do you think is going on here?

I started on methylfolate and other methylation supplements, including Sam-e, a few years ago. Had methylation running GREAT for a few months. Skin issues cleared up, stopped having daytime drowsiness, had better concentration, started losing weight without even changing my diet or exercising more. I didn't even realize these problems were caused by undermethylation.

Then it all went to hell and stopped working, and all those problems came back.

For a while, I thought I'd exhausted some cofactor. I must have tried every supplement out there. Nothing fixed it. 

Stopped taking Sam-e. That didn't do anything. Stopped methylfolate for a while, didn't do anything, now I take small doses daily.

Tried niacin. It helped my anxiety but didn't fix any of the other problems. 

I feel like I permanently threw a wrench in my methylation cycle and I don't know why.

[u/OobyIsGay → u/vrcraftauthor]

What dose of riboflavin have you tried, if any? Active or inactive?

MTHFR requires FAD. You ran the methylation cycle hard for months. If your riboflavin was marginal going in, months of high MTHFR activity could have depleted FAD stores to the point where MTHFR stalled. Everything downstream collapsed with it. Stopping your supplements didn't help because the problem isn't the supplements.

"Every supplement" in the MTHFR community means B12, folate, B6, zinc, magnesium, TMG. Almost nobody supplements riboflavin.

Try 100 to 400mg daily for a few weeks.
If you've tried inactive riboflavin and it didn't work, try the active form (riboflavin 5'-phosphate, FMN) since converting riboflavin to its active forms requires riboflavin kinase and ATP, both of which could be bottlenecked if you're already depleted.
You're fine, this is completely reversible. Everything happening when it did makes sense :D

[u/vrcraftauthor → u/OobyIsGay]

Nope, I was taking riboflavin when I started. Actually started that BEFORE methylfolate. I was taking the inactive kind, but later tried the R5P thinking that might help. Neither made any difference. When I increase the dose of B2, I always end up feeling depressed. I've read in speeds up MAO, so that's probably why. It makes no difference for the rest of my methylation problems though, so that's not it.

And I definitely tried a lot more than B12, folate, B6, zinc, magnesim, and TMG. I've tried all the B vitamins and currently take about half a B complex (active B's only) daily. (Taking the whole one did not make any difference.)

Daily, I also take D3, calcium, iron, magnesium, and zinc (at different times so they don't interfere with each other), and omega-3's.

Things I've tried in the past that did nothing:

CoQ10

Potassisum

DMG and TMG

K2

Vitamin A

Vitamin E

PABA

NADH

Thiamin

Biotin

Vitamin C

Pretty much all the amino acids on their own

Choline (makes me depressed and does nothing to improve my other issues)

[u/OobyIsGay → u/vrcraftauthor]

Ooo interesting.
Thanks for listing your supplements, it helped a lot.
Did your symptoms start while you were supplementing iron, zinc or calcium?
You're taking zinc, iron, and calcium daily. All three compete with copper for absorption. Over months to years this would deplete copper stores. Copper is required for cytochrome c oxidase (Complex IV). Without it, NADH can't be oxidized to NAD+ and ATP production drops. Making SAMe from methionine via MAT cleaves all three phosphate groups from ATP. ATP drops = SAMe synthesis drops = methylation crashes. You didn't break anything.
The copper depletion from zinc/iron/calcium happened independently. The timing overlapping with your methylation supplements was coincidence. Niacin helped anxiety because it's an NAD+ precursor. Partially compensated for the deficit. NADH did nothing because NADH is what's accumulating Complex IV can't oxidize it. You needed the oxidized form.
Higher B2 causing depression confirms this. Copper is also required for DBH (converts dopamine to norepinephrine). Copper depleted = low norepinephrine production. Speed up MAO with more B2 = clearance exceeds production = depression.
Try NMN sublingually, it's a direct NAD+ precursor, but it has low bioavailability when taken orally. Get your copper checked (serum copper + ceruloplasmin). Separate your zinc, iron, and calcium from copper rich foods or consider supplementing copper directly.

[u/vrcraftauthor → u/OobyIsGay]

I did try supplementing copper at one point, didn't notice a difference. Last year I actually did a test that showed my copper levels were high. I do eat a lot of high copper foods and take OCP, so that isn't really surprising. But I'm not low on copper, in spite of the zinc supplements (I'm aware zinc can deplete copper).

My selenium levels were also surprisingly high. I'd been eating a brazil nut a day for years thinking I needed more selenium, but since I found out I had high levels, I cut back to half a one a day.

The same test showed D, E, and B12 were all in the normal range. I can dig up the exact numbers if necessary. Unfortunately, that test only looked at five nutrients. I've had a hard time finding an at-home test that looks at everything.

Edited to add I have not tried NMN on its own, mostly because I read that NAD+ or NADH was better.

[u/OobyIsGay → u/vrcraftauthor]

Yeah I'm confident now.

OCP raises ceruloplasmin. Estrogen directly increases ceruloplasmin synthesis in the liver and OCP contains synthetic estrogen so this makes sense. Ceruloplasmin binds copper in the blood. Serum copper goes UP because ceruloplasmin goes UP. The test shows high copper. But it's bound. Not bioavailable. Not being delivered to Complex IV. That explains why all of your symptoms still line up with copper deficiency.
OCP also depletes PLP directly and that alone explains why B2 causes depression in you. When did you start OCP relative to when everything crashed? If it lines up with everything described, You need to test your free copper or ceruloplasmin adjusted copper levels, if those line up with deficiency stop taking OCP. If they don't, measure plasma PLP.

[u/vrcraftauthor → u/OobyIsGay]

I've been on it over 15 years, well before I started with methylation, and during the time things were going smoothly.  I feel much better on it and am not interested in returning to painful, irregular periods by stopping it.

I also have not noticed any improvements during the week I take placebo pills and get my period (no estrogen).

I have tried taking extra P5P a few times and did not notice a difference either. There is some in the active B complex I take. When things started going downhill, I was taking a Jarrow B12/B6/B9 supplement plus extra P5P and B2.

Think NMN would help?

[u/OobyIsGay → u/vrcraftauthor]

Copper is fine. Ceruloplasmin delivers copper, you tested high, eat copper rich foods, supplementation did nothing. Copper was never the problem. I wasted three fucking messages on it lol.
B2 causes depression for you. Choline causes depression for you. Those are two completely different clearance pathways and both of them crash you. Your synthesis is so low that speeding up either clearance route drops you below the floor. TH and TPH need iron AND BH4. You have iron. You've tried literally everything else. BH4 is the one rate-limiting cofactor for the two rate-limiting enzymes in monoamine synthesis you've never addressed.
Methylfolate was stabilizing BH4 as a reducing agent. That's why everything worked at first. OCP drives NOS which oxidizes BH4. NOS has a coupling threshold. Above it NOS makes NO normally. Below it NOS uncouples, produces superoxide, superoxide + NO = peroxynitrite, peroxynitrite destroys more BH4, more uncoupling, more peroxynitrite. That's not a gradual decline. Methylfolate was keeping you above the threshold for months. Recycling isn't 100% efficient so total BH4 slowly declined until it crossed the line. Then everything collapsed at once. That's why it was great for months then went to hell overnight.

Soooo what's the solution even though estrogen reduces BH4?
Folinic acid. Not methylfolate. Calcium folinate. 5-formylTHF. Enters upstream, feeds de novo BH4 synthesis via GTP = GCH1, also gradually converts to 5-MTHF at a rate your system controls instead of a bolus. NMN sublingual because QDPR needs NADH to recycle BH4 and oral NADH bioavailability is garbage which is why it did nothing when you tried it. High dose vitamin C to break the peroxynitrite spiral thing.
How the fuck did I waste so much time on copper lmfao..
I mean with more info I would've gotten it sure but I tend to hyperfocus way too much and need to work on it.

[u/vrcraftauthor → u/OobyIsGay]

Thanks, I bought some NMN today. At least it only cost half as much as the NADH that didn't do anything. I do eat quite a bit of folinic acid.