N,N-dimethyltryptamine (DMT) regulates adult neurogenesis in vitro/ vivo

I wonder how much neurogenesis affects our recovery,  because benzo recovery aswell as other psychiatric drugs recovery must englobe more that neurogenesis itself,  but

This is a very interesting article adressing neurgenesis , as neurogenesis in adults ,  worth to read about it i guess that even if you have a difficult time to read (like me) its worth  of an effort to read,

I separated and highlighted some small parts of the article for all to take a look ,

Please  DownloadIng the pdf. file, makes it much easier to read !

This Scientific article of N,N-dimethyltryptamine (DMT) compound found in the hallucinogenic tea ayahuasca, regulates adult neurogenesis in vitro and in vivo

https://www.nature.com/articles/s41398-020-01011-0

Abstract

N,N-dimethyltryptamine (DMT) is a component of the ayahuasca brew traditionally used for ritual and therapeutic

purposes. Here, we have examined, in vitro and vivo, the potential neurogenic effect of DMT.

Our results demonstrate that DMT administration activates the main adult neurogenic niche, the subgranular zone of the dentate gyrus of the hippocampus, promoting newly generated neurons in the granular zone. Moreover, these mice performed better, compared to control non-treated animals, in memory tests,which suggest a functional relevance for the DMT-induced new production of neurons in the hippocampus.

Interestingly, the neurogenic effect of DMT appears to involve signaling via sigma-1 receptor (S1R) activation since S1R

antagonist blocked the neurogenic effect. Taken together, our results demonstrate that DMT treatment activates the

subgranular neurogenic niche regulating the proliferation of neural stem cells, the migration of neuroblasts, and

promoting the generation of new neurons in the hippocampus, therefore enhancing adult neurogenesis and

improving spatial learning and memory tasks.

Introduction

N,N-dimethyltryptamine (DMT) is a natural compound

found in numerous plant species and botanical preparations, such as the hallucinogenic infusion known as ayahuasca1 classified as a hallucinogenic compound that

induces intense modifications in perception, emotion, and

cognition in humans2–4

. DMT is present in several animal tissues, such as the lung and brain being considered as an endogenous trace neurotransmitter with different physiological roles, including neural signaling and brain/peripheral immunological actions7–10.

  DMT is also present in human blood, urine, and cerebrospinal fluid11–13. Furthermore, some evidence suggests that DMT can be sequestered into and stored in the vesicle system of the brain and that environmental stress increases its levels in mammals’ central nervous system (CNS)14–16. DMT binds and exerts an agonist activity on subtypes 1A and 2A of the serotonin receptor (5-HT)17,18. These receptors are Gprotein-coupled receptors (GPCRs) belonging to the family of serotonergic receptors and are involved in numerous cascades of intracellular signaling, with high expression in several regions of the CNS.

Some studies have demonstrated that DMT also binds with low affinity to nonserotonergic receptors, such as the sigma-1 receptor (S1R).

The S1R, traditionally thought to be an opioid receptor, is

now classified as a highly conserved transmembrane protein member of an orphan family and located mainly in the

membrane of the endoplasmic reticulum.

σR‐1 is widespread in the CNS, mainly in the prefrontal cortex, hippocampus, and striatum19. Interestingly, in mammals, one of the natural endogenous ligands of the σR-1 is DMT14.

This receptor has been associated with several cellular

functions, including the brain, such as lipid transport,

metabolism regulation, cellular differentiation, signaling (in

response to stress), cellular protection against oxidant

agents, myelination and,  most recently, neurogenesis20–24. Neurogenesis is the process of generating new functional neurons, mainly in the SVZ and the subgranular zone of the DG of the hippocampus. In mammals,

This process occurs mostly during the prenatal period, being

significantly reduced in adults25–30. In humans, although

the presence of adult neurogenesis has been recently reported during aging31–33,

most of the studies indicate that there are no substantial evidence to support it. A recent review by Duque and Spector suggests that, in adult age, preservation of the existing neurons is more important in contrast to the generation of new ones34.

Neurogenesis is a complex process involving multiple

cellular activities including the proliferation of neural stem cells (NSC; progenitors), migration and differentiation, survival, acquisition of cell fate and maturation, and integration of these newly born neurons in existing neuronal circuits

  All these processes are precisely regulated by multiple factors35. Advancements in the knowledge of these factors and their mechanism of action could help us to investigate possible new instruments that will allow us to expand the limited endogenous neurogenic capacity of the adult brain and, consequently, opening new fields for the development of effective therapies in the treatment of brain damage and neurodegenerative diseases.

Neurodegenerative diseases (including Parkinson’s,

Alzheimer’s, Hungtinton’s, etc.) and acute neural damage

(such as stroke and traumatic brain injury) are characterized by a gradual and selective loss of neurons in the

affected regions of the nervous system. One common

feature in these disorders is an impairment in the proliferation of progenitor cells in the neurogenic niches36,37.

In animal models reproducing the pathological hallmarks

of Alzheimer disease, a loss of neurogenic capacity has

been described in the SVZ38. This decrease is also

observed in the postmortem brains of Parkinson’s

patients, suggesting that the loss of neurogenic activity is

due to the loss of dopamine, affecting the neural precursors in the adult39. These data support the fact that in

neurodegenerative diseases, such as Alzheimer’s and

Parkinson’s, not only degeneration and death of mature

neurons occur but also the process of formation of new

neuronal progenitors in the adult brain is negatively

affected. According to these data, the stimulation of

endogenous populations of stem cells and neuronal progenitors could be a promising approach to improve the

functionality of some of the regions affected by neurodegenerative pathologies. In fact, the stimulation of neurogenesis has already been proposed as a new therapeutic strategy for psychiatric and neurological diseases40–44, and several studies have reported that the

clinical efficacy of antidepressant drugs is frequently

linked to the capacity of these drugs to induce neurogenesis45–48.

Based on the data above mentioned including our

results on the potent neurogenic effect of the other

components of the Ayahuasca49, the main objective of this

work was to analyze the possible role of DMT in adult

neurogenesis, as well as to elucidate its mechanism of

action.

Results

DMT controls the stemness of neural progenitors in vitro

through the S1R

We first analyzed whether the sigma-1 receptor (S1R)

was expressed on murine NSCs isolated from the subgranular zone of the dentate gyrus of the hippocampus.

Figure 1a shows S1R expression on neurospheres in the

basal state determined by immunocytochemistry and

western blot analysis. To analyze the “stemness” of cultured neurospheres, we determined the expression of

potentiality markers of this state. Then, we performed WB

analysis after treatment of these cultures during 7 days

under proliferative conditions (see “Materials and methods”) with DMT alone or in combination with the different antagonists. Our results (Fig. 1b) show significant

reductions in protein levels of musashi-1, nestin, and

SOX-2 in the SGZ-derived neurospheres after treatment

with DMT, suggesting a loss of stemness in NSCs in the

NS cultures. When these cultures were pre-treated with

BD1063, a specific antagonist for S1R, this effect was

reversed, and stemness marker levels were similar to those

observed in basal conditions. On the contrary, the

expression of stemness markers in those cultures treated

with DMT together with the mixed serotonin 5-HT1A/

2A receptor antagonist methiothepin, the selective

5-HT2A receptor antagonist ritanserin, or the selective

5-HT1A receptor antagonist WAY100635, significantly

decreased stemness as occurred in DMT-treated cultures.

These results suggest that DMT promotes a loss of

“stemness” or and undifferentiated state of the neurospheres, through the S1R.

DMT promotes the proliferation in vitro of NSCs  neural stem cells (NSC; progenitors),

Other NS cultures were used to study proliferation;

thereby, the number and diameter of the neurospheres

were evaluated.

DMT notably increased thenumber and size of the neurospheres in NS cultures after 7 days of treatment, indicating that DMT promotes the proliferation of adult hippocampal-derived neural progenitors. DMT proliferative effect was blocked when cultures were treated with BD1063 showing a significant decrease in the number and size of neurospheres, similar to basal conditions.

Also, significant differences, in number and size of neurospheres, were observed when cultures were treated with DMT combined with methiothepin, ritanserin, or WAY100635.. We next analyzed changes in two well-known markers for proliferation, ki67 and proliferating cell nuclear antigen (PCNA) (Fig. 1d, e). Fluorescent immunocytochemical analysis of ki67 expression (Fig. 1d) showed an increase in the number of ki67+ cells in the NS after treatment with DMT,

suggesting a direct effect of DMT on the proliferation ability of NSCs. This effect was clearly reverted when cultures were also incubated with the antagonist BD1063 (BD). Similar results were obtained by western blot analysis and subsequent quantification of PCNA (Fig. 1e). No significant differences in the expression of ki67 and PCNA were observed when cultures were preincubated with other DMT antagonists.

These results indicate that DMT stimulates in vitro, through the S1R, the proliferation of neural progenitors of the adult neurogenic niche of the hippocampus.

Discussion

We have previously described that β-carbolines alkaloids, the three main alkaloids present in Banisteriopsis

caapi and harmol, the main metabolite of harmine in

humans, play an important role as key regulators on

adult neural stem cell activity49. Using an in vitro model

of adult neurogenesis, we showed that they promote the

proliferation and migration of progenitor cells and

induced their differentiation mainly into a neuronal

phenotype. The main limitation to that work was that

the potential role of DMT, other active compounds

contained in ayahuasca brews, was not described.

Moreover, previous studies performed on rodents and

primates55–58, and more interestingly in humans4,59,

suggest that ayahuasca infusion has antidepressant

activity, a therapeutic effect usually linked to hippocampal neurogenesis. This work extends our previous

results indicating the role of DMT, one of the main

compounds of the hallucinogenic infusion ayahuasca, in

adult neurogenesis.

Our results in vitro and in vivo show that DMT is a key

regulator in the activity of adult NSCs neural stem cells (NSC; progenitors),, since this compound plays an important role in regulating the expansion and differentiation of the stem cell population located in the SGZ, one of the main adult neurogenic niches. This is revealed in vitro by an increase in the number and size of primary neurospheres and an increased expression of ki67and PCNA, which indicates a high rate of proliferation and loss of stemness after treatment with DMT. Increased proliferation does not indicate neuronal commitment60;

however, DMT also induced an increase in β-III-tubulin+

and MAP-2 + cells, suggesting promotion of differentiation toward a neuronal phenotype and increasing the total

numbers of the neuron that reach neuronal maturity.

Interestingly, in contrast to that previously described on

the action of carbolines in vitro49, we have also found an  increase in the number of other neural cells such as

astrocytes and oligodendrocytes after DMT treatment.

Similar results were observed in vivo, with an increased

proliferation rate of the NSCs and a larger population of

doublecortin expressing neuroblasts migrating to the

hippocampal granular layer to generate new neurons.

Moreover, these have a functional impact since DMT

treatment during 21 days clearly improved mouse performance in learning and memory tasks, in which the

hippocampus is considered to play an essential role. These

observations are in agreement with previous works

showing that adult hippocampal neurogenesis plays an

important role in these cognitive functions61–65.

Considering these effects, we can determine that the DMT has the capacity to regulate the expansion and destination of

stem cell populations and therefore contribute to memory

and learning processing in the dentate gyrus.

Neurogenesis consists of proliferation and loss of stemness of the NSCs, migration of neuroblasts and differentiation into functional neurons. Results here obtained

demonstrate that DMT controls all these stages. Interestingly, in addition to the neurogenic potential, DMT also induced the formation of astrocytes and oligodendrocytes.

This ability for controlling neurogenesis is of great interest,

since in pathological conditions, the renewal of the neurons

must be optimized by acting simultaneously on several

processes40,66. We have previously indicated that many

molecules67–71 and recently β-carbolines contained in ayahuasca49 exerted and effect on cell proliferation and differentiation, therefore the effect of DMT stimulating cell

proliferation and differentiation is not exclusive to this

compound.

One of the goals of this work is that additionally to its neurogenic effect, DMT also stimulated migration and

new generation of astroglial cells and oligodendrocytes,

what highlights the versatility of this compound as it can

promote all the processes involved in full adult neurogenesis.

Specifically, astrocytes are known to support the proliferation, survival, and maturation of developing neurons

and neuroblasts that have already committed to neuronal

lineages, But also to promote neurogenesis73,74.

In fact, previous works demonstrated that astrocytes in vitro could be directly converted into neurons or stem-like cells, pointing to the plasticity of these somatic glial cells75–77. No previous studies on the neurogenic effect of DMT have

been described, but in comparison with the effect of other

ayahuasca components such as β-carbolines49, we can

conclude that the effect of DMT on adult neurogenesis is

considerably more potent.

  As an additional value to the generation of neurons, the glial cells formation induced by DMT might be an ideal target for in vivo neuronal conversion after neural injury, since some studies have achieved to generate proliferating, non-tumorigenic neuroblasts from resident astrocytes78.

The main therapeutical implication of the results here obtained is derived from the close relationship between neurogenesis and antidepressant activity described in several animal models79.

DMT is considered a serotoninergic drug because its

mechanism of action consists in agonism at different

serotoninergic receptors, especially the 5-HT2A receptors

widely described as inducers of neurogenesis80, but also

psychedelics (reviewed by Dos Santos and Hallak81).

One of the main limitations that arise when designing a

possible drug from the results obtained is to achieve the

desired neurogenic effect without causing the patient

hallucinogenic effects secondary to treatment with DMT,

through the activation of 5-HT2A receptors.

The results here obtained indicate that the observed effects of DMT are mediated by the activation of the S1R. In this regard, it has been shown that the stimulation of the S1R by different agonists enhances neurogenesis in the hippocampus23,82.

  Moreover, in vivo evidence suggests that the

σ1R deficiency interrupts the adult neurogenesis22. In

humans, the use of S1R agonists, such as fluvoxamine,

shows its involvement in neuroplasticity83, suggesting an

important role in improving learning mechanism. In

clinical studies, some S1R agonists, including fluvoxamine, donepezil, and neurosteroids, improve cognitive impairment84,85

Adult hippocampal neurogenesis iswidespread in mammals, including humans, and may act as a key regulator in cognition, memory, and emotionrelated behavior86.

Deficits in adult neurogenesis are associated with the physiopathology of depression and modulation of neurogenesis is behind the action of several

antidepressants79.

Recently, a previous study has described the role of other psychoactive tryptamine, 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT), in neurogenesis87. In contrast to the intracerebral injection of 5-MeO-DMT administered by these authors, we used DMT i.p. that can cross the blood–brain–barrier, which facilitates its future administration in humans. Moreover, the neurogenic effect of DMT through the S1R activation is combined to the antagonism of 5-HT2A receptor,

avoiding the hallucinogenic effects of these tryptamine

derivatives. This information could be very useful for

the future development of new treatments against

neurodegeneration.

In conclusion, this study shows that DMT present in the

ayahuasca infusion promotes neurogenesis by stimulating

the expansion of neural progenitors populations, and by

inducing the differentiation of these NSCs. Moreover, the

neurogenic stimulation observed after DMT treatment

correlates with an improvement in spatial learning and

memory tasks in vivo.

Stimulation of the neurogenic niches of the adult brain can contribute substantially to the antidepressant effects of ayahuasca in recent clinical

studies. The versatility and complete neurogenic capacity

of the DMT guarantee future research regarding this

compound. In addition, its ability to modulate brain

plasticity indicates its therapeutic potential for a wide

range of psychiatric and neurological disorders, among

which are neurodegenerative diseases.

I hope that some folks/buddies could give their time to read and express their opnions and point of view on this subject

The preparation of the drink contains crushed stalks of the Banisteriopsis caapi (mariri or jagube) vine with the leaves of the Psychotria viridis (queen or chacrona) shrub, undergoing a decoction (cooking) of the plants  concentrated is prepared with only the two plants (jagube and chacrona), in the same decoction process as any other Ayahuasca. What makes the difference is only the method of preparation that takes place in a larger and more detailed process

The decoction (cooking) time is longer, in this process of uninterrupted boiling and the continuous mixing of the components with water until a thicker consistency occurs, acidity is eliminated (because it contains 0% acidity, it does not make you feel bad, or if it occurs cleaning it is subtle without unpleasant symptoms).In this process of liquid evaporation and addition of Jagube and Chacrona until the tea becomes well concentrated (12×1), Ayahuasca Pura allows for a smaller dosage in the consecration (20ml) without compromising the final effect (safe dose that allows for an experience). intense and special.

  there are buddies around here who tried this aproach themselves,  i hope those could chime in to share  their experience with ayahuaca(dmt) 

I can't read all that lol. But I have read buddies having good success with DMT and claimed it healed them. I believe there's definitely something in using psycodelics to help heal the brain.

Here's 1 success story

http://www.benzobuddies.org/forum/index.php?topic=246876.msg3142483#msg3142483

and another who had success with it

http://www.benzobuddies.org/forum/index.php?topic=261657.msg3304470#msg3304470

Not sure if my brain could take it, but I wouldn't rule it out completely

I can't read all that lol. But I have read buddies having good success with DMT and claimed it healed them. I believe there's definitely something in using psycodelics to help heal the brain.

 

Here's 1 success story

http://www.benzobuddies.org/forum/index.php?topic=246876.msg3142483#msg3142483

 

and another who had success with it

http://www.benzobuddies.org/forum/index.php?topic=261657.msg3304470#msg3304470

  downloading Turns it to be much easier to read,

  i highlighted some interesting parts above, its a lot of reading (hard to read) but very interesting, that said folks with a better cognition could bring us their views, a psychedelic experience is not pleasant in this state, but probably not unpleasent too, thanks for posting this links supporting,

im looking forward to try this aproach,  using a this tea once a month, or  once every 6.months? ( im not aware of how its done) its better that a setback due to any other neurotoxic agent,

  cheers mate!