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Offlinerungi
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psilocybin loacation
    #7533041 - 10/18/07 10:40 PM (13 years, 6 months ago)

Does anyone know the location in individuals cells where psilocyin is made and stored? Fungi cell walls seems to be composed of various glucans and chitin.


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Re: psilocybin loacation [Re: rungi]
    #7537473 - 10/20/07 12:05 AM (13 years, 6 months ago)

i always thought it was all over in the mycelium and mushrooms except the spores.


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Offlinemulege
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Re: psilocybin loacation [Re: LayYouIn]
    #7560465 - 10/25/07 09:38 PM (13 years, 6 months ago)

I think you misunderstood his question... He means what individual part of the cell, i.e. mitochondria, cell walls, etc.


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Re: psilocybin loacation [Re: mulege]
    #7560992 - 10/25/07 11:59 PM (13 years, 6 months ago)

Quote:

mulege said:
I think you misunderstood his question... He means what individual part of the cell, i.e. mitochondria, cell walls, etc.




:confused:

sorry, i cheated my way through biology.  :wink:


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Re: psilocybin loacation [Re: LayYouIn]
    #7561921 - 10/26/07 08:22 AM (13 years, 6 months ago)

I would assume in the Golgi complex and in the vacuoles. They are made jointly in the ribosomes and modified in the Rough E.R. Just like every other amide/protein.


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Re: psilocybin loacation [Re: Pooter]
    #7566219 - 10/27/07 11:53 AM (13 years, 6 months ago)

Quote:

Pooter said:
I would assume in the Golgi complex and in the vacuoles. They are made jointly in the ribosomes and modified in the Rough E.R. Just like every other amide/protein.



:nonono:
Go elsewhere with your disinformation.
This is just bullshit in its purest form. Seriously, I have seldom heard anything quite this horribly incorrect. There isn't an grain of truth or knowledge behind this.
Many proteins are synthesized in the cytoplasm others are made in the Mitochondria. Amides are made in pretty much every part of the cell. Proteins are mainly synthesized in the ER neither psilocin nor psilocybin are proteins though. Rather: they are biosynthesized by enzymes(which are proteins).

I haven't been able to find any clear explanations where the biosynth of these alkaloids happen but the best bet would be the cytoplasm as they are not intended for excretion.


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Re: psilocybin loacation [Re: rungi]
    #7569569 - 10/28/07 03:27 PM (13 years, 6 months ago)

Quote:

Nalim said:
Proteins are mainly synthesized in the ER



When I took biochemistry and cell biology at university, we were clearly taught that protein synthesis (with the exception of mitochondrial proteins) starts in the cytoplasm where ribosomes translate mRNA into peptides. But of course, the complete synthesis of a particular protein (eg. with sugar moieties or the like) usually entails subsequent modification in the ER and other organelles.

As to the intracellular location of psilocybin, I'd assume that cytoplasmic enzymes carry out the synthesis. The storage would likely be in secretory vesicles (if secreted), or possibly freely in the cytoplasm, depending on what the organism uses the alkaloids for. The presence of a cell wall does not hinder secretion: Many fungi and bacteria secrete antibiotics/toxins. If secreted (to the intercellular space), the alkaloids could perhaps have a signalling function like hormones or neurotransmitters do.

Quote:

Nalim said:
they are not intended for excretion.



How do you know?

Fungal cells don't intend anything, neither does the mycelium nor the mushroom. Fungi don't think, therefore they don't intend. Animals can intend. Psilocybes presumably can't. The same goes for nature and natural selection. I know that it's common for people to see intention everywhere in nature, but that's simply incorrect.

With the relative lack of research having been done in this field, I can't see any reason to assume that psilocin/-cybin is not secreted by individual fungal cells. The molecules certainly resemble other signalling molecules (serotonin) which human neurons secrete abundantly.

Some people like to think that the alkaloids are synthesized solely for sentient beings to cherish the mushrooms, thereby ensuring the survival of the genome (= intention). This, however, doesn't explain its function in prehistoric times before the emergence of sentience, so I don't buy it. Whatever their function, the alkaloids must give the organism a selective advantage. If not, those without alkaloid synthesis would quickly gain an advantage by being more energy efficient.

Btw rungi, why the interest in the location? Are you developing a new alkaloid extraction method?
:cool:


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Offlinerungi
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Re: psilocybin loacation [Re: Hyoscyamus]
    #7577181 - 10/30/07 05:11 PM (13 years, 6 months ago)

No, but that is my interest, and it comes from a book
There is a book called Fungal Cell Wall by Jose Ruiz Herrera In it are some protocol for extracting component of cells and cell walls using glass beads,ultrasonic oscillators centrifuges,French presses, and various other techniques. There is no mention of alkaloids and or antibiotics (are they different?)being components in cell walls. My guess is that psilocyin is made and stored in a similar location that penicilin is. Maybe it has a similar biological function as well. I do not know how penicilin is extracted and purified from fungal cells.

The location of penicilin synthesis and storage? I think this has been studied in great detail because of the profits high yields produce.


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Re: psilocybin loacation [Re: rungi]
    #7577981 - 10/30/07 08:43 PM (13 years, 6 months ago)

Penicilin is made from the metabolites, not the fungi itself.
RR


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Re: psilocybin loacation [Re: rungi]
    #7580628 - 10/31/07 02:48 PM (13 years, 6 months ago)

In my biochemistry course we used ultrasound to disintegrate cell membranes, whereafter the substrate was centrifuged so we easily could extract certain wanted cell contents (in our case rat liver cell mitochondrial inner membrane proteins :tongue:). This process could be used for a wide range of things, but there must be better ways of extracting psilocybin. If psilocybin is in fact secreted, one would benefit immensely by in some way triggering the fungal cells to secrete, thereby being able to both collect the alkaloid and keep the fungal cells alive after delivery. Otherwise you'd have to break open the cells or extract otherwise (methanol?).


Quote:

rungi said:
There is no mention of alkaloids and or antibiotics
(are they different?)being components in cell walls.



An alkaloid is a molecule of a certain structure (an amine) and origin (mostly plants, but also fungi, dream fish, bamboo worms, ...) which often has some effect on humans/animals, whereas an antibiotic is a toxin secreted against competing organisms (eg. bacteria) with the "purpose" to kill or inhibit.

The main function of a cell wall is to prevent the cell within from bursting due to high pressure. That's why penicillin kills some bacteria - the cell wall components are prevented from assembling and therefore the cell bursts. But even though the cell wall keeps the cell from bursting, it's not completely impenetrable; substances that the cell secretes are able to pass through. Some bacteria even have a capsule on the outside of the cell wall which prevents antibiotics from reaching the wall and do damage.


Quote:

rungi said:
My guess is that psilocyin is made and stored in a similar location
that penicilin is. Maybe it has a similar biological function as well.



If "meant" for secretion (like antibiotics), psilocybin would most likely be found in secretory vesicles more or less continually flowing from the golgi apparatus towards the cell surface and either promptly expelled or aggregated right beneath the plasma membrane waiting for the cell to be triggered so that a large quantum can be discharged simultaneously (like human insulin).

Nobody(?) knows the function of psilocybin (or the other psilocybian alkaloids), so it could serve some function in the cell wall, however I seriously doubt it. I can't see what function a small molecule like psilocybin would serve there. Other fungi and bacteria (both with cell walls) live quite happily without psilocybin being present anywhere.

At first glance psilocybin sure looks like a neurotransmitter, but once you test a substance on a different (not too closely related) organism "anything" could happen. Some substances are harmless to rats, but deadly toxic to humans, and vice-versa. So it could have an antibiotic function on some competing organism in nature which psilocybian mycelia often encounter, but the molecule just resembles a neurotransmitter so much that I'd personally put my money on a kind of cell-to-cell communication.

:alert: Like molecules often have like functions, even in very distant cousins.


Quote:

rungi said:
I do not know how penicilin is extracted and purified from fungal cells.
The location of penicilin synthesis and storage? I think this has been studied in great detail because of the profits high yields produce.



The original extraction method can hardly be a secret (no original patents effective), but might be very heavy reading. But remember: Psilocybin is not penicillin, so don't assume too much. Rather, find some scientific articles on the subject. Try searching PubMed online and then get the articles from a research library. A quick search gave me this article title: "A novel extraction procedure for psilocybin and psilocin determination in mushroom samples". Maybe that's what you're looking for?

Extraction of psilocybin from mushroom culture is supposedly inefficient, which is the reason Sandoz makes it synthetically (Indocybin). But of course, "inefficient" may simply be in the eyes of a corporation which must focus on the bottom line. It may perhaps not be too silly if one wants to "play around" with it hobbywise. I've thought about it myself.


Quote:

RogerRabbit said:
Penicilin is made from the metabolites, not the fungi itself.



I assume you mean medically used penicillins? The substance secreted by the Penicillium molds does by itself inhibit bacterial cell wall synthesis (thereby killing the offender). But the stuff used in medicine today is a wide range of analogs, so in that sense "medical penicillins" are metabolites of the fungal secretion.

I don't think many doctors would dare inject a pure fungal extraction into a severely ill patient today - even though old remedies such as maggots are getting a rennaisance in medicine. :syringe: :syringe: :syringe: :shocked:


Edited by Hyoscyamus (10/31/07 03:00 PM)


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Re: psilocybin loacation [Re: Hyoscyamus]
    #7580672 - 10/31/07 03:03 PM (13 years, 6 months ago)

extracting psilocybin is not that hard.

just grind up mushrooms, and place the powder in methanol over steady heat no more than 65c and constant stirring in a reflux apparatus simple condenser.

Once completed filter the liquid thru filter paper (or under suction thru a filter core).

evaporate most of the methanol off over low heat (65c) and then allow to cool, first in a refrigerator, then in a freezer. This will allow the crystals of psilocybin to form. filter them out while still cold and re-warm/cool the leftover methanol to allow for a second recrystallization


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InvisibleHyoscyamus
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Re: psilocybin loacation [Re: Slimz]
    #7580877 - 10/31/07 03:57 PM (13 years, 6 months ago)

Methanol, yes. But in what range of purity would you guess the resulting extract would be? Many other substances would be extracted along with psilocin and psilocybin in this manner.

Of course, none of them would be more toxic than the mushrooms themselves, but for the true nerd high purity of the sought chemical is vital. :mushroom2:

Is methanol easy to acquire in the US? It isn't here, you need a "poison permit". :thumbdown:


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Re: psilocybin location [Re: rungi]
    #7581033 - 10/31/07 04:27 PM (13 years, 6 months ago)

Let's keep to the subject at hand please, and not let it degenerate into another extraction thread. Thanks.
RR


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Re: psilocybin loacation [Re: Hyoscyamus]
    #7585377 - 11/01/07 09:55 PM (13 years, 6 months ago)

Quote:

Hyoscyamus said:
Quote:

Nalim said:
Proteins are mainly synthesized in the ER



When I took biochemistry and cell biology at university, we were clearly taught that protein synthesis (with the exception of mitochondrial proteins) starts in the cytoplasm where ribosomes translate mRNA into peptides. But of course, the complete synthesis of a particular protein (eg. with sugar moieties or the like) usually entails subsequent modification in the ER and other organelles.

As to the intracellular location of psilocybin, I'd assume that cytoplasmic enzymes carry out the synthesis. The storage would likely be in secretory vesicles (if secreted), or possibly freely in the cytoplasm, depending on what the organism uses the alkaloids for. The presence of a cell wall does not hinder secretion: Many fungi and bacteria secrete antibiotics/toxins. If secreted (to the intercellular space), the alkaloids could perhaps have a signalling function like hormones or neurotransmitters do.

Quote:

Nalim said:
they are not intended for excretion.



How do you know?

Fungal cells don't intend anything, neither does the mycelium nor the mushroom. Fungi don't think, therefore they don't intend. Animals can intend. Psilocybes presumably can't. The same goes for nature and natural selection. I know that it's common for people to see intention everywhere in nature, but that's simply incorrect.

With the relative lack of research having been done in this field, I can't see any reason to assume that psilocin/-cybin is not secreted by individual fungal cells. The molecules certainly resemble other signalling molecules (serotonin) which human neurons secrete abundantly.

Some people like to think that the alkaloids are synthesized solely for sentient beings to cherish the mushrooms, thereby ensuring the survival of the genome (= intention). This, however, doesn't explain its function in prehistoric times before the emergence of sentience, so I don't buy it. Whatever their function, the alkaloids must give the organism a selective advantage. If not, those without alkaloid synthesis would quickly gain an advantage by being more energy efficient.

Btw rungi, why the interest in the location? Are you developing a new alkaloid extraction method?
:cool:




This is all way over my head, just doing some reading in the wrong section i guess :wink:  I had the thought Maybe the production of alkaloids from the mushrooms somehow attract species of insects which eat on, and even reproduce on mushrooms. At some point maybe this insect is then attracted to manure in whatever form.  This would help would help the spread of the mushrooms and ensure their overall survival.  Anyway it might be possible that insects could be attracted to certain types of mushies because both benefit each other ( i know ive had loads of maggots living inside my wild mushies before ) and the key to this attraction being the production of said alkaloids?


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Re: psilocybin loacation [Re: Myconut]
    #7638527 - 11/15/07 09:30 AM (13 years, 5 months ago)

Many things are possible and your example of spreading of the mushrooms by insects is indeed common in nature. But remember that insect species aren't the same across the world, and that not all psilocybian mushrooms are associated with manure. There are also very large environmental differences between Thailand, Germany, and Oregon.

The alkaloids, however, are the same (although differing in content) in the mushrooms found in these diverse locations, so their function must be associated with something found in every single geographical location where you find psilocybian mushrooms, for else the process of natural selection will "weed out" those strains producing unneeded alkaloids since this won't be energy efficient.

Now, what function could these alkaloids possibly have in such diverse locations? My own bet is one inside the mushrooms themselves, but it could of course also be something external, perhaps symbiotic with other organisms as you point out yourself. Parasitic organisms often have quite crazy life cycles, and perhaps psilocybin and the other alkaloids are a part in some strange interaction between the mushrooms and other organisms in their environment.

The maggots that you may find in wild Psilocybes don't necessarily get something that they can't get anywhere else, but they may have a preference for them over other mushrooms.


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Re: psilocybin loacation [Re: Hyoscyamus]
    #7642184 - 11/16/07 12:20 AM (13 years, 5 months ago)



I took the above picture in a microscopy class that I attended last week. One of my classmates brought a "Psilocybe cyanofriscosa" specimen; this is a tiny gill fragment which has been thoroughly crushed under the cover slip to release the cystidia. The crushing caused the tissue to blue quite a bit. There is no dye, all the bluing is caused by psilocin degradation. I noticed that some areas were much bluer than other areas and thought of this thread as I took a picture. It was very obvious that the psilocin is not equally distributed in the mushroom tissue - it looks like the cells had organelles which were so dark blue that they were almost opaque.


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Re: psilocybin loacation [Re: Alan Rockefeller]
    #7642265 - 11/16/07 12:40 AM (13 years, 5 months ago)

Quote:

Alan Rockefeller said:

...There is no dye, all the bluing is caused by psilocin degradation...




That is an awesome pic, Alan.

*Heads for POTM thread*


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Re: psilocybin loacation [Re: Alan Rockefeller]
    #7643024 - 11/16/07 07:30 AM (13 years, 5 months ago)

Interesting. However, the thorough crushing dispersing and oxidizing the psilocin makes things difficult to identify. Wherever psilocin is stored it will be free from contact with oxygen, so there might be an aggregation of (non-blue) vesicles in the cytoplasm, most probably requiring a dye of some kind to be added.

Would it be possible to get a picture of lesser crushed "au naturel" non-gill tissue, dyed for contrast? Or has someone perhaps got some hi-res TEM pics?

Btw, has "cyanofriscosa" gotten a proper name yet?
:confused:


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Re: psilocybin loacation [Re: Hyoscyamus]
    #7643479 - 11/16/07 11:43 AM (13 years, 5 months ago)

The reason you see blue in that one spot is because that's the only part that is in focus. Microscopes have a narrow depth of field range, and as you can see, the only spores visible are also in that same area with the bluing. There's also hundreds, if not thousands of individual cells in that picture which I estimate at around 100X to 200X.
RR

Who has any proof that blue is caused by psilocin? What about non-active mushrooms that also bruise when damaged?


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Re: psilocybin loacation [Re: RogerRabbit]
    #7646493 - 11/17/07 02:51 AM (13 years, 5 months ago)

> The reason you see blue in that one spot is because that's the only part that is in focus. Microscopes have a narrow depth of field range, and as you can see, the only spores visible are also in that same area with the bluing.

The fresh gill fragment was very small in the first place and was crushed between the cover slip and slide by putting a LOT of force on a pencil eraser. We are looking at a very thin layer here, it wasn't focusing in on different cells at different depths. It looked similar to the naked eye - you could clearly see blue spots.

> There's also hundreds, if not thousands of individual cells in that picture which I estimate at around 100X to 200X.

It is at 400X and there are closer to dozens of cells in view. You can see parts of some cystidia near the center.

> Who has any proof that blue is caused by psilocin?

All dark spored mushrooms which bruise blue contain psilocin, making me think that the psilocin and bluing are very closely related.

> What about non-active mushrooms that also bruise when damaged?

They all have light colored spores.

> Btw, has "cyanofriscosa" gotten a proper name yet?

No.

> That is an awesome pic, Alan.
> *Heads for POTM thread*

Thanks, I really wish I had taken a macro shot of the microscope slide with my camera - it was really beautiful.

> Would it be possible to get a picture of lesser crushed "au naturel" non-gill tissue, dyed for contrast?

I don't have any samples, this was at a weekend microscopy class with a whole bunch of people. A shroomery member showed up with one tiny very blue psilocybe.

> Or has someone perhaps got some hi-res TEM pics?

I hope to have some in a few months.


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