One for the geeks. This is an article from O'shaughnessy's journal of cannabis in clinical practice. I am chucking it in for future reference and search facility...I hope i'm in the right forum

Winter/Spring 2005

O'Shaughnessy's
Journal of the California Cannabis Research Medical Group


"Retrograde Messengers": Scientific American Explains how Cannabinoids Work

"Marijuana has clear medicinal benefits," Roger A. Nicoll and Bradley N. Alger state unambiguously in an article in the December 2004 Scientific American. "Marijuana alleviates pain and anxiety. It can prevent the death of injured neurons. It suppresses vomiting and enhances appetite--useful features for patients suffering the severe weight loss that can result from chemotherapy."

Nicoll is a professor of pharmacology at UCSF, Alger a professor of physiology and psychiatry at the University of Maryland School of Medicine. Their article, entitled “The Brain’s Own Marijuana,” focuses on mechanism of action. Its clear, expository style and illuminating graphics are in the magazine’s finest tradition. A lengthy excerpt follows.

In 1964, after nearly a century of work by many individuals, Raphael Mechoulam of the Hebrew University in Jerusalem identified delta-9-tetrahydrocannabinol (THC) as the compound that accounts for virtually all the pharmacological activity of marijuana. The next step was to identify the receptor or receptors to which THC was binding.
Receptors are small proteins embedded in the membranes of all cells, including neurons, and when specific molecules bind to them—fitting like one puzzle piece into another—changes in the cell occur. Some receptors have water-filled pores or channels that permit chemical ions to pass into or out of the cell. These kinds of receptors work by changing the relative voltage inside and outside the cell. Other receptors are not channels but are coupled to specialized proteins called G-proteins. These G-protein-coupled receptors represent a large family that set in motion a variety of biochemical signaling cascades within cells, often resulting in changes in ion channels.
In 1988 Allyn C. Howlett and her colleagues at St. Louis University attached a radioactive tag to a chemical derivative of THC and watched where the compound went in rats’ brains. They discovered that it attached itself to what came to be called the cannabinoid receptor, also known as CB1. Based on this finding and on work by Miles Herkenham of the National Institutes of Health, Lisa Matsuda, also at the NIH, cloned the CB1 receptor [located the gene that encodes it].
The importance of CB1 in the action of THC was proved when two researchers working independently—Catherine Ledent of the Free University of Brussels and Andreas Zimmer of the Laboratory of Molecular Neurobiology at the University of Bonn—bred mice that lacked this receptor. Both investigators found that THC had virtually no effect when administered to such a mouse: the compound had nowhere to bind and hence could not trigger any activity. (Another cannabinoid receptor, CB2, was later discovered; it operates only outside the brain and spinal cord and is involved with the immune system.)
As researchers continued to study CB1, they learned that it was one of the most abundant G-protein coupled receptors in the brain. It has its highest densities in the cerebral cortex, hippocampus, hypothalamus, cerebellum, basal ganglia, brain stem, spinal cord and amygdala. This distribution explains marijuana’s diverse effects. Its psychoactive power comes from its action in the cerebral cortex. Memory impairment is rooted in the hippocampus, a structure essential for memory formation. The drug causes motor dysfunction by acting on movement control centers of the brain. In the brain stem and spinal cord, it brings about the reduction of pain; the brain stem also controls the vomiting reflex. The hypothalamus is involved in appetite, the amygdala in emotional responses. Marijuana clearly does so much because it acts everywhere.
Over time, details about CB1’s neuronal location emerged as well. Elegant studies by Tamás F. Freund of the Institute of Experimental Medicine at the Hungarian Academy of Sciences in Budapest and Kenneth P. Mackie of the University of Washington revealed that the cannabinoid receptor occurred only on certain neurons and in very specific positions on those neurons. It was densely packed on neurons that released GABA (gamma-aminobutyric acid), which is the brain’s main inhibitory neurotransmitter (it tells recipient neurons to stop firing). CB1 also sat near the synapse, the contact point between two neurons. This placement suggested that the cannabinoid receptor was somehow involved with signal transmission across GABA-using synapses. But why would the brain’s signaling system include a receptor for something produced by a plant?
The same question had arisen in the 1970s about morphine, a compound isolated from the poppy and found to bind to so-called opiate receptors in the brain. Scientists finally discovered that people make their own opioids—the enkephalins and endorphins. Morphine simply hijacks the receptors for the brain’s opioids.
It seemed likely that something similar was happening with THC and the cannabinoid receptor. In 1992, 28 years after he identified THC, Mechoulam discovered a small fatty acid produced in the brain [arachidonic acid ethanolamine, AEA] that binds to CB1 and that mimics all the activities of marijuana. He named it anandamide, after the Sanskrit word ananda, “bliss.” Subsequently, Daniele Piomelli and Nephi Stella of the University of California at Irvine discovered that another lipid, 2-arachidonoyl glycerol (2-AG), is even more abundant in certain brain regions than anandamide is. Together the two compounds are considered the major endogenous cannabinoids, or endocannabinoids. (Recently investigators have identified what look like other endogenous cannabinoids, but their roles are uncertain.) The two cannabinoid receptors clearly evolved along with endocannabinoids as part of natural cellular communication systems. Marijuana happens to resemble the endocannabinoids enough to activate cannabinoid receptors.
Conventional neurotransmitters are water-soluble and are stored in high concentrations in little packets, or vesicles, as they wait to be released by a neuron. When a neuron fires, sending an electrical signal down its axon to its tips (presynaptic terminals), neurotransmitters released from vesicles cross a tiny intercellular space (the synaptic cleft) to receptors on the surface of a recipient, or postsynaptic, neuron. In contrast, endocannabinoids are fats and are not stored but rather are rapidly synthesized from components of the cell membrane. They are then released from places all over the cells when levels of calcium rise inside the neuron or when certain G-protein-coupled receptors are activated.
As unconventional neurotransmitters, cannabinoids presented a mystery, and for several years no one could figure out what role they played in the brain. Then, in the early 1990s, the answer emerged in a somewhat roundabout fashion. Scientists (including one of us, Alger, and his colleague at the University of Maryland School of Medicine, Thomas A. Pitler) found something unusual when studying pyramidal neurons, the principal cells of the hippocampus. After calcium concentrations inside the cells rose for a short time, incoming inhibitory signals in the form of GABA arriving from other neurons declined.
At the same time, Alain Marty, now at the Laboratory of Brain Physiology at the René Descartes University in Paris, and his colleagues saw the same action in nerve cells from the cerebellum. These were unexpected observations, because they suggested that receiving cells were somehow affecting transmitting cells and, as far as anyone knew, signals in mature brains flowed across synapses in one way only: from the presynaptic cell to the postsynaptic one.
It seemed possible that a new kind of neuronal communication had been discovered, and so researchers set out to understand this phenomenon. They dubbed the new activity DSI, for depolarization-induced suppression of inhibition. For DSI to have occurred, some unknown messenger must have traveled from the postsynaptic cell to the presynaptic GABA-releasing one and somehow shut off the neurotransmitter’s release.
“Such backward, or ‘retrograde,’ signaling was known to occur only during the development of the nervous system. If it were also involved in interactions among adult neurons, that would be an intriguing finding—a sign that perhaps other processes in the brain involved retrograde transmission as well. Retrograde signaling might facilitate types of neuronal information processing that were difficult or impossible to accomplish with conventional synaptic transmission. Therefore, it was important to learn the properties of the retrograde signal. Over the years, countless molecules were proposed. None of them worked as predicted.
Then, in 2001, one of us (Nicoll) and his colleague at the University of California at San Francisco, Rachel I. Wilson—and at the same time, but independently, a group led by Masanobu Kano of Kanazawa University in Japan—reported that an endocannabinoid, probably 2-AG, perfectly fit the criteria for the unknown messenger. Both groups found that a drug blocking cannabinoid receptors on presynaptic cells prevents DSI and, conversely, that drugs activating CB1 mimic DSI. They soon showed, as did others, that mice lacking cannabinoid receptors are incapable of generating DSI. The fact that the receptors are located on the presynaptic terminals of GABA neurons now made perfect sense. The receptors were poised to detect and respond to endocannabinoids released from the membranes of nearby postsynaptic cells.
Over time, DSI proved to be an important aspect of brain activity. Temporarily dampening inhibition enhances a form of learning called long-term potentiation—the process by which information is stored through the strengthening of synapses. Such storage and information transfer often involves small groups of neurons rather than large neuronal populations, and endocannabinoids are well suited to acting on these small assemblages. As fat-soluble molecules, they do not diffuse over great distances in the watery extracellular environment of the brain. Avid uptake and degradation mechanisms help to ensure that they act in a confined space for a limited period. Thus, DSI, which is a short-lived local effect, enables individual neurons to disconnect briefly from their neighbors and encode information.
A host of other findings filled in additional gaps in understanding about the cellular function of endocannabinoids. Researchers showed that when these neurotransmitters lock onto CB1 they can in some cases block presynaptic cells from releasing excitatory neurotransmitters. As Wade G. Regehr of Harvard University and Anatol C. Kreitzer, now at Stanford University, found in the cerebellum, endocannabinoids located on excitatory nerve terminals aid in the regulation of the massive numbers of synapses involved in coordinated motor control and sensory integration. This involvement explains, in part, the slight motor dysfunction and altered sensory perceptions often associated with smoking marijuana.
Recent discoveries have also begun to precisely link the neuronal effects of endocannabinoids to their behavioral and physiological effects. Scientists investigating the basis of anxiety commonly begin by training rodents to associate a particular signal with something that frightens them. They often administer a brief mild shock to the feet at the same time that they generate a sound. After a while the animal will freeze in anticipation of the shock if it hears the sound. If the sound is repeatedly played without the shock, however, the animal stops being afraid when it hears the sound—that is, it unlearns the fear conditioning, a process called extinction. In 2003 Giovanni Marsicano of the Max Planck Institute of Psychiatry in Munich and his co-workers showed that mice lacking normal CB1 readily learn to fear the shock-related sound, but in contrast to animals with intact CB1, they fail to lose their fear of the sound when it stops being coupled with the shock.
The results indicate that endocan-nabinoids are important in extinguishing the bad feelings and pain triggered by reminders of past experiences. The discoveries raise the possibility that abnormally low numbers of cannabinoid receptors or the faulty release of endogenous cannabinoids are involved in post-traumatic stress syndrome, phobias and certain forms of chronic pain. This suggestion fits with the fact that some people smoke marijuana to decrease their anxiety. It is also conceivable, though far from proved, that chemical mimics of these natural substances could allow us to put the past behind us when signals that we have learned to associate with certain dangers no longer have meaning in the real world.

The article goes on to describe some of the roundabout strategies that university and drug-company scientists are employing to develop a drug that doesn’t produce the unwanted side effects of THC. It concludes, “In a remarkable way, the effects of marijuana have led to the still unfolding story of the endocan-nabinoids. The receptor CB1 seems to be present in all vertebrate species, suggesting that systems employing the brain’s own marijuana have been in existence for about 500 million years. During that time, endocannabinoids have been adapted to serve numerous, often subtle, functions. We have learned that they do not affect the development of fear, but the forgetting of fear; they do not alter the ability to eat, but the desirability of the food, and so on. Their presence in parts of the brain associated with complex motor behavior, cognition, learning and memory implies that much remains to be discovered about the uses to which evolution has put these interesting messengers.




RETROGRADE SIGNALING is the mechanism by which the body's won cannabinoids work. In top diagram GABA (an inhibitory neurotransmitter) and glutamate (an excitatory neurotransmitter) are being released from presynaptic nerve cells, crossing the synapse (gap between cells) and hitting the postsynaptic cell below. If the endocannabinoid 2-AG is not present, the GABA will inhibit the postsynaptic cell's calcium level tripper production of 2-AG, the endocannabinoid will hit on the CB1 receptor of the presynaptic cell, halting the release of GABA, thus allowing the excitatory sinals to activate the postsynaptic cell (bottom diagram)..