Sigma Xi, The Scientific Research Society

Mechanisms of Action of LSD: The study of serotonin-containing neurons in the brain may provide the key to understanding drug-induced hallucinations and their relationship to dreams and psychosis Author(s): Barry L. Jacobs and Michael E. Trulson Source: American Scientist, Vol. 67, No. 4 (July-August 1979), pp. 396-404 Published by: Sigma Xi, The Scientific Research Society Stable URL: http://www.jstor.org/stable/27849328 . Accessed: 29/01/2014 06:32
Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at . http://www.jstor.org/page/info/about/policies/terms.jsp

.
JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org.

Sigma Xi, The Scientific Research Society is collaborating with JSTOR to digitize, preserve and extend access to American Scientist.

http://www.jstor.org

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

Barry L. Jacobs Michael E. Trulson

Mechanisms

of Action

of LSD

The study of serotonin-containing neurons in the brain may provide the key to understanding drug induced hallucinations and their relationship to dreams and psychosis

"Last Friday, April 16,1943,1 was forced to stopmy work in the laboratory in the middle of the afternoon and to go home, as I was seized by a peculiar restlessness associated with a sensation of mild dizzi

later, in an attempt to verify that the to episode was indeed attributable the ingestion of LSD, Hofmann took what he thought would be a small ness. Having I lay down reached home, it a and sank in kind of drunkenness which quantity of the drug, 250 fig.As turned out, this was approximately was not unpleasant and which was char five times the dose necessary to pro acterized by extreme activity of imagi in an av nation. As I lay in a dazed condition with duce intense hallucinations my eyes closed (I experienced daylight as erage adult male. His personal ac count of the drug's effects was re disagreeably bright) there surged upon me an uninterrupted stream of fantastic since virtually accurate, markably images of extraordinary plasticity and everything he described has been vividness and accompanied by an intense, confirmed by subsequent laboratory kaleidoscope-like play of colors. This studies with large numbers of subjects condition gradually passed offafter about under controlled conditions. two hours."

acid diethylamide(LSD). Three days

neurotransmitter that appears to play a key role in the action of LSD. More generally, we will consider brain se rotonin in the broad context of its role inmammalian behavior. The discussion of the mechanism of action of LSD also illustrates the revolution that has occurred during the past two decades in our under standing of the action of a variety of psychoactive drugs, such as cocaine, and chlorpromazine amphetamine, (the most frequently prescribed an tipsychotic). Although these drugs act a number of different through share the common mechanisms, they property of altering chemical neuro transmission in the brain. Therefore, our story logically begins with a brief review of chemical neurotransmis
sion.

can be divided This description, from the journal of The effects of LSD three into (2): so categories general a Albert Hofmann (1), Swiss chemist matic weak symptoms?dizziness, working in Basel during World War ness, nausea, tremors, creeping or II, heralds the dawn of the era of ex sensations on the skin, and tingling perimental study of hallucinogenic blurred symp vision; perceptual experience re drugs, forHofmann's and colors, vi toms?altered shapes sulted from his accidental ingestion sual hallucinations, (a synesthesia of an unknown quantity of d-lysergic mixing of senses, such as the trans formation of sounds into changes in visual perception), and a distorted time sense; affective and cognitive and rapid mood symptoms?large Barry Jacobs, Associate Professor of Psychol in thinking, de difficulty changes, and a member of ogy at Princeton University and dreamlike feel personalization, in the interdepartmental program graduate The small amount ofLSD needed at ing. received his UCLA doctorate neuroscience, to produce these profound psycho and was a postdoctoral fellow in the psychia at Stanford. In 1977-78 he was try department logical effects makes it the most po a visiting scientist at the Salk Institute. His tent psychoactive drug known, on a
interests include the neural bases of behavior, animal models complex mammalian of human psychopathology, psychoactive who re Trulson, drugs, and sleep. Michael research microgram-for-microgram basis.

The human brain is comprised of tens of billions of nerve cells, or neurons. Neurons bring information to the brain concerning the current status of, or the occurrence of any change in, the body's internal milieu and the external world. Neurons also call into action many of the glands and mus cles of the body. By acting upon each other, inways we are just beginning to neurons the understand, provide tremendous integrative capacity of the brain human that underlies memory, thought, and emotion. The action of neuron upon neuron is out by means of small carried amounts of chemicals released from the terminal endings of the neurons into the minute gaps, or synapses, between them (Fig. 1). These mole cules then cross the synapse and im pinge upon the dendrites or the cell body of the receiving neuron and produce either excitation or inhibi

in the area of brain neurochemistry primarily and behavior. Address for Professor Jacobs: in Neuroscience, Program Department of

ceived his doctorate in biopsychology from the been at in 1974, has University of Iowa Princeton since then as a lecturer and director of the neurochemistry laboratory in the psy chology department. He is currently an Alfred P. Sloan research interests are fellow. His

NJ 08544.

Psychology,

Princeton

University,

Princeton,

the past fifteen years, re During search in the field of neuroscience has greatly elucidated the neurobiological that mediate LSD's mechanisms effects. The primary psychological purpose of this article is to describe the research that has led to our of these present understanding mechanisms. Much of the discussion will be centered on serotonin, a brain

396

American

Scientist,

Volume

67

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

was found to be present in the mam malian central nervous system (CNS) in significant quantities and to be in varying amounts in concentrated different regions of the brain. This led to the proposal that serotonin was a CNS neurotransmitter. In the twenty-five years since that time, an impressive body of evidence has been to indicate that seroto accumulated nin does indeed act as a central neu
rotransmitter.

contact with an Figure 1. One neuron makes other not by actually touching but by com across minute called sy municating spaces, of chemical neurotransmit napses, by means ters. The

classical synapse is between the axon terminal of one neuron and the dendrites or cell body of another neuron. Fig. 2 shows details of synaptic neurotransmission.

tion. If the summated effect of the hundreds, or even thousands, of in puts that a neuron receives favors excitation and reaches a threshold level, the cell will "fire" and transmit an electrical impulse down its axon. the impulse reaches the ter When minal portion of the axon and causes the release of some small proportion of the chemical neurotransmitter stored there, the entire cycle begins
again.

Structurally, the serotonin molecule is similar to a portion of the larger in that they both con LSD molecule tain an indole nucleus. Based on this structural similarity, Gaddum, in working in England, and Wooley, the United States, independently proposed that LSD might act to block the synaptic action of serotonin in the brain (3, 4). They buttressed their argument by demonstrating that LSD exerted a powerful blocking effect on serotonin's action in peripheral tissue studied in vitro. This hypothesis at tracted a great deal of attention, be cause, in addition to being considered a hallucinogenic was drug, LSD a to be thought by many prototypic one psychotomimetic drug?i.e. whose effects mimicked psychosis. Thus, the argument went, a key ele ment in the etiology of psychosis might be either decreased amounts of brain serotonin or the synthesis of an that anta endogenous compound serotonin's in the action gonized
brain.

postsynaptic
Figure

neuron

2. An axon terminal of a serotonin containing neuron (presynaptic neuron) makes synaptic contact with one of its target neurons

the neuron). (postsynaptic L-tryptophan, amino acid precursor of serotonin, is brought to the neuron by the blood. Serotonin is syn thesized from tryptophan inside serotonergic nerve terminals and is stored in packets called vesicles. When an action potential invades the axon terminal, the vesicles release their con tents into the synaptic gap and bombard the neuron to produce either excita postsynaptic tion or inhibition. Serotonin is inactivated by being taken back up into the terminal, where it is catabolized oxidase (MAO) by monoamine to form acid 5-hydroxyindoleacetic (5

HIAA).

LSD

and serotonin:
overview

This ambitious hypothesis was soon shaken by the report that brom-LSD, which is LSD with a single bromine atom attached, although as effective as LSD in blocking serotonin's action on peripheral tissue, was devoid of potent psychic effects in humans (5). It was therefore unlikely that the serotonin

serotonin

indole nucleus Figure 3. The serotonin molecule is similar hallucinogenic in Fig. 6). drugs,

structure of the to that of several (as shown including LSD

Historical

For many years scientists had known of a blood-borne chemical that pro duced vasoconstriction (a serum factor that affected blood vessel tonus, hence the name serotonin) and of a substance present in the gut that caused intestinal motility. In the mid-twentieth century, serotonin, the both single compound producing these effects, was isolated and syn structure thesized, and itsmolecular elucidated as 5-hydroxytryptamine. Soon thereafter, in 1953-54, serotonin

could account for its hal lucinogenic action, since this ability was shared by a close analog of LSD that was nonhallucinogenic. It is worth noting, however, that both sides of this argument were based on studies of LSD's action on peripheral tissue. Because of the general inac cessibility of brain tissue, there had been very little direct test of the central effects of LSD. The remain der of this paper will be concerned with examining these latter studies, but because so many of them involve brain serotonin, we must first provide

abilityofLSD to block the action of

some background information about this neurotransmitter.

Basics

of brain serotonin

Serotonin is a small and relatively molecule that is synthesized simple from the essential amino acid tryp tophan. After a meal, tryptophan is transported, by the blood, from the neurons, gut to serotonin-containing where serotonin synthesis takes place (Figs. 2 and 3). The newly formed serotonin is stored in the axon ter

1979

July-August

397

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

minai in packets called vesicles. When an action potential invades the axon terminal, the serotonin molecules are into the synaptic gap and released bombard the serotonin receptors on or target, neuron. the postsynaptic, Inactivation of serotonin's synaptic is accomplished action primarily through a process by which the mol ecules are taken back into the axon is destroyed by terminal. Serotonin the enzyme monoamine oxidase, and the resulting major catabolite of se acid rotonin, 5-hydroxyindoleacetic in brain (5-HIAA), can be measured and cerebrospinal tissue, urine, fluid. In the mid-1960s, a seminal series of studies employing the newly devel oped technique of fluorescence his tochemistry detailed the localization of the cell bodies and axon terminals of the CNS neurons that utilize sero tonin as their transmitter (6, 7). brain tissue is treated with When gas and then para-formaldehyde with ultraviolet bombarded light, serotonin-containing neurons give off a yellow fluorescence (Fig. 4). This technique revealed that CNS seroto nin was localized almost exclusively within neurons of the raphe nuclei (small clusters of cells on the midline of the brainstem) and their axon ter minals. The total number of these neurons, at least in the rat brain, where they have been studied most than is no more intensively, 10,000-20,000. Thus, they represent a very small proportion of all CNS

are neurons 4. Serotonin-containing Figure made visible through the use of fluorescence of the In this photomicrograph histochemistry. of the rat, dorsal raphe nucleus in the midbrain each oval yellow spot is the cell body of a sero neuron; the dark areas at the tonin-containing bottom on both sides are large fiber tracts fasciculi). The photomi (medial longitudinal 1mm of brain crograph covers approximately tissue sity.) courtesy from top to bottom. (Photomicrograph Yale Univer of George Aghajanian,

LSD
Modern

and serotonin:
era

Fortunately, because of the pioneer studies employing ing anatomical this fluorescence histochemistry, hypothesis could be directly exam ined by recording the electrical ac tivity of serotonergic (i.e. serotonin in cell bodies localized containing) stem of the in the brain clusters tight in rat. In 1968, George Aghajanian into the region serted microelectrodes of the dorsal raphe nucleus of anes thetized rats and, after obtaining a stable baseline sample of the cell's activity, he intravenously adminis tered a low dose of LSD (10). Nor neurons these of the discharge mally, rats is slow (1-2 in anesthetized spikes/second) and regular. Following the injection of LSD, as hypothesized, these serotonergic cells displayed an abrupt and complete cessation of ac tivity. By contrast, the activity of neighboring nonserotonergic neurons was either unaffected or slightly in In a subsequent creased by LSD. that reported study, Aghajanian even in much brom-LSD, higher doses, had a much smaller effect on (11). serotonergic neurons than LSD Finally, through the use ofmicroion tophoresis, which permits the direct application of small amounts of sub stances to single neurons, Aghajanian that LSD demonstrated depressed the activity of serotonergic neurons through an effect directly on the cell body and had little direct effect on any of the other CNS neurons that were studied (12).

of raphe LSD's The first report that LSD had a sig specific depression neuro neuronal activity, in conjunction with effect on brain nificant the uniformly in 1961 transmission was published inhibitory synaptic action of serotonin in the forebrain, The serotonin-containing neurons (or by Daniel Freedman (8). He found a disinhibition of raphe in that a single injection of LSD produces raphe neurons) have widely ramifying neurons. or Be over in are serotonin axons that often sent out creased the level of brain postsynaptic, target, cause of the densest in 24 rat while brom those the For distances. aggregations percent, by example, great these target neurons are in areas of in even higher doses, failed to the posterior portion of the brainstem LSD, the brain that mediate processing of send their axons down the length of affect brain serotonin. Thus, although or emotive visual ef in had similar the dor LSD and brom-LSD the spinal cord, and those information, we an for ex mechanism have obvious on in of the the nuclei serotonin fects sal and median periphery, raphe their central effects were significantly anterior brainstem send their axons plaining the major affective, percep different. In an important extension into various portions of the forebrain. tual, and cognitive effects of LSD. that LSD it is hypothesized In the forebrain, raphe neurons most of this study, Freedman and his col Thus, of seroto acts to the vi to in of the that addition innervate activity depress portions leagues reported heavily neurons, in brain serotonin, LSD increases which, sual system and portions of the limbic nin-containing through disinhibition, cause a release system, a group of structures known produced significant decreases in the to be important in emotional experi brain level of the major metabolite of of activity of neurons in the visual Al ence and expression system, the limbic system, and many 5). serotonin, 5-HIAA (9). These findings (Fig. other brain areas (13). This model de to most led the that LSD these anatomical of hypothesis might though tails have been worked out in the rat,% inactivate or depress the activity of does not preclude the possibility that increase in LSD may also exert a direct action on serotonin neurons. The they have relevance to humans be cause the pattern of distribution of brain serotonin was attributed to its other brain neurons, including sero tonin target cells. that within neurons serotonin in the central nervous sys accumulation were no longer releasing itdue to their tem is fairly constant across a variety The argument that a significant pro ofmammalian inactivity. species.
neurons. 398 American Scientist, Volume 67

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

portion of LSD's psychic effects can be attributed to its action on sero tonergic neurons is buttressed by the fact that other hallucinogens which produce similar psychic effects, such as psilocin, N,N dimethyltryptamine and 5-methoxy DMT (5 (DMT), also have an indole nu MeODMT), cleus structure (Fig. 6) and depress raphe unit activity (11, 14). On the other hand, psychoactive drugs that elicit different psychic effects, such as (THC?the A-tetrahydrocannabinol or active component of marijuana) neither have the indole amphetamine, nucleus structure nor suppress the of serotonin-containing activity neurons. The few human experiments that bear on this issue also provide indirect support for the serotonin hypothesis. For example, drugs that decrease brain serotonin levels pot in humans, entiate the effects of LSD and drugs that increase brain sero tonin levels decrease the effects.

hippocampus

-PPP
amygdala

...MFB

Figure 5. The axons of serotonin-containing over great distances. neurons often project of cell bodies Clusters (dots) of serotonin neurons, both the dorsal and me containing dian raphe nuclei as well as the group of cells labeled B-9, are shown in cross section through

of the rat. Axons from these cell the midbrain into the forebrain to make con bodies ascend

tact with structures such as the lateral genic ulate nucleus (LGNV, a portion of the visual and amygdala system) and the hippocampus (portions of the limbic system). The medial is one of the major forebrain bundle (MFB) the midbrain pathways connecting raphe neurons with their forebrain target cells.

the need to impute an major parameters of the drug's effects in in humans (e.g. development of tol of state, similarity underlying It was against this backdrop of re volves using some aspect of animal erance). search that we began our own studies behavior (or physiology) as a model of of LSD and, more generally, the brain the human variable. Such models are We chose the cat as our experimental serotonin system. We felt that a animal because of its vast behavioral founded on the assumption that their number of important issues remained validity can be established through repertoire and the ease with which to be resolved. Since we are ulti that changes in the any behavioral changes could be ob demonstrating studies mately interested in behavior, would served. Although animal directly parallel previous changes in the effects of hallucinogenic drugs on humans. The had examined the effects of LSD and changes need not be on behavior in or even analogous, but related hallucinogens serotonergic neurons be the same in homologous, a variety of species, few of them had freelymoving animals as they were in merely must covary systematically. anesthetized or immobilized animals? explored a complete dose range or On a microgram-for-microgram basis, the behavioral examined In order to qualify as an animal model closely so much more potent why is LSD for the actions of hallucinogenic drugs important, none of changes. Most than other hallucinogens? What role, a particular them had reported an effect that was behavior in humans, if any, do other neurotransmitter to be specific to LSD. would have to change specifically in demonstrated systems play in the action of halluci in our initial study, we response to this class of drug and no Accordingly, the to cats and at LSD nogenic drugs? What mediates administered other, vary in frequency ormagnitude in in a be rapid and dramatic diminution manner, dose-dependent tempted to compile a complete and effects that fol LSD's psychological detailed record of all major behavioral elicited by drug doses within the Is human lows its repeated administration? range, and closely parallel the changes, with special attention to in activity of brain se the decrease rotonin neurons that is produced by LSD by any normal approximated condition? physiological which obviates

An animal model forLSD

of many of these issues Resolution necessitated examining the behav ioral effects of LSD. Since the use of human subjects in such experiments is precluded for ethical reasons, we turned to animal experiments. How ever, using animals in studies dealing with variables like hallucinations, which are based exclusively on self the unanswerable report, raises question of how to discern what a is feeling, nonverbal organism or Another tack, thinking, perceiving.

drugs LSD, psiFigure 6. The hallucinogenic locin, and DMT (N,N-dimethyltryptamine)

have an indole nucleus structure similar to that of the serotonin molecule (Fig. 3). 1979 July-August 399

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

behaviors

cifically byLSD.

that might be evoked spe

We discovered that, in addition to increased frequency of head and body shakes, grooming, investigatory be re havior, and hallucinatory-like not two other behaviors, sponses, previously reported, were observed

thesis inhibitor, the behavioral effects of the two drugs are synergistic (18). Therefore, besides helping to estab lish the validity of themodel, the data from these drug studies also buttress the serotonin hypothesis of halluci nogenic drug action. In an attempt to extend the useful ness of thismodel to known actions of in humans, we ob hallucinogens served something surprising. If an adult human is given an effective dose

Limb flicks and abortive 16). grooming increased in frequency in direct relation to the dose of LSD (beginning from a baseline of essen tially zero in saline-treated animals and progressively increasing in ani mals treated with 2.5, 10, 25, and 50

underLSD (15, with highprobability

scratch) or performs it in midair. as the cat begins to Apparently, and groom, it becomes distracted never finishes the grooming sequence. Whether the limb flicks, abortive head and body shakes, and grooming, the like actually represent halluci nations is irrelevant, since the model as simply employs these measures to in hu hallucinations parallels
mans.

again on days 2,3, and 4, there will be a marked decrease in the effective ness of the drug in producing psychic ofbodyweight).Limb flicking change by the second day, and an al Mg/kg is a species-specific behavior seen in most complete loss of effectiveness by normal cats almost exclusively in re day 4 (19, 20). This effect is called a of to the these studies, tolerance. Paralleling presence sponse foreign we gave LSD to cats in an intermedi substance, such as water, on the paw. The paw is lifted and rapidly and repetitively shaken or snapped out ward from the body (Fig. 7). In abor tive grooming, the cat orients to the body surface as if to groom but does not perform the consummatory response (bite, lick, or grooming

on day 1 and then ofLSD (1 Mg/kg)

One of the most intriguing aspects of these studies was the fact that the of the peak behavioral magnitude was effect of LSD significantly than that greater produced by psilo serotonin cin, DMT, 5-MeODMT, receptor blockade, or serotonin de pletion. For example, a 50 fig/kgdose of LSD produced an average of ap proximately 40 limb flicks per hour, whereas the other drugs, regardless of dose, typically produced 5-10 limb flicks per hour. Since all of these drugs, including LSD, were known to block serotonin neurotransmission, we reasoned that the magnitude of LSD's behavioral effect must be at tributable to some additional action. Therefore, we began to explore the possibility that, in addition to sero tonin, other neurotransmitters might be involved. in several other laboratories Studies had indicated that LSD also acted to mimic of the neuro the action transmitter dopamine (22, 23). We confirmed this directly with electro studies that examined physiological the effect of LSD on the activity of dopaminergic neurons in the rat brain (24). Furthermore, using a simple behavioral model in the rat,we found that another very potent hallucino genic drug, DOM (2,5-dimethoxy-4 also had a methylamphetamine), effect, halluci 5 and nogens DMT, psilocin, were virtually devoid of MeODMT action (25). Since a dopaminergic previous study had reported that also depressed the activity of DOM significant dopaminergic whereas the indole nucleus that DOM might be as hypothesized as LSD in our effective behaviorally cat model. This was confirmed with behavioral studies, inwhich we found that DOM produced approximately 40 limb flicks per hour (17). Thus, themost potent hallucinogenic drugs may be those that both inacti vate brain serotonin and mimic brain Serotonin inactivation dopamine. may be necessary and sufficient for hallucinogenesis (psilocin, DMT, and 5-MeODMT have serotonergic, but no dopaminergic, action), while the action may modulate dopaminergic the amplitude of the effect. This is supported by clinical evidence. When patients who are having "bad trips" on LSD are given antipsychotic drugs, which are potent dopamine-receptor blockers (e.g. chlorpromazine), they
serotonin-containing neurons, we

or other cats are given LSD, Figure 7. When related they display drugs, hallucinogenic in that are seen exclusively several behaviors to these drugs. One of these is the response limb flick, which is seen in normal cats only in response to the presence of a foreign substance on the paw. The paw is raised from the ground and then rapidly shaken or flicked away from the body.

specificity of these behavioral changes is indicated by the fact that they are never seen in response to single injections ofmany other classes of psychoactive drugs, such as THC, caffeine, atropine (an amphetamine, and chlorphenira anticholinergic), mine (an antihistaminic). Nor were they seen in response to brom-LSD, the nonhallucinogenic relative of LSD, or tryptamine, a nonhallucino genic indole nucleus compound. Most important, however, these behaviors were elicited by hallucinogens that are structurally to LSD related and psilocin) (DMT, 5-MeODMT, and known to depress the activity of neurons (17). serotonin-containing They were also elicited by a drug that blocks serotonin's action on its target neurons and by a drug that decreases brain levels of serotonin by inhibiting its synthesis. Furthermore, when is administered LSD to cats previ ously treated with a serotonin-syn
400 American Scientist, Volume 67

The

dose on day 1 and then administered an additional 50 Mg/kg dose on day 2 (21). Much to our surprise, the single 10 Mg/kg pretreatment produced a nearly complete blockade of the be havioral effects of the 50 Mg/kg dose, and the single 50 Mg/kg pretreatment made the second 50 Mg/kg dose as ineffective as an injec behaviorally tion of saline. In subsequent behav ioral studies we found that this tol erance to LSD, which was complete 24 hours after the initial injection, actually had begun to develop within two hours after the injection, at a time when the drug itselfwas still exerting its primary effect!We shall return to this finding later, when we discuss research on themechanism mediating
tolerance.

or a high (50 Mg/kg) ate (10 Mg/kg)

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

in the typically report a diminution a but of the intensity experience of the hallucinatory continuation activity. The dopaminergic action of hallucinogenic drugs might also be relevant to the fact that they are fre to mimic psy quently considered chosis (26), since the preponderance of current biochemical evidence bearing on schizophrenia indicates an overactivity in the brain dopamine of a consideration system. Thus, action of hallucinogenic dopaminergic drugs, in addition to their action on brain serotonin, seems to explain to be what previously appeared somewhat anomalous and disparate findings.

MN
t ........ Xgg

behavior

Serotonin

neurons

and

vil* was devised to Figure 8. A special apparatus record the electrical activity of single neurons in awake, freely moving animals. A cat is placed in a large, soundproofed box that is electrically can be con shielded. The animal's behavior tinuously monitored through a one-way chamber. Various and recorded by videotape in one wall of the mirror such as gross electrodes, as well as the the EEG, those for recording are attached to a standard microelectrodes, connector and the entire assembly is secured to the skull with an acrylic. During an experi ment, the cat is connected to various amplifiers and a polygraph machine by means of a flexible cable attached to the connector on the animal's head.

The next step in our research was a crucial one for us. We were interested in recording the activity of seroto neurons in freely nin-containing so cats that behavior could be moving studied concomitantly. Over the past ten years, in conjunction with Dennis at and Ronald McGinty Harper we re have and UCLA, developed fined a technique first used by James it involves recording Olds. Basically, of single neuron activity by means bundles of insulated microwires that can be advanced through the brain in small steps by an attached mechani elec cal microdrive (Fig. 8). These trodes differ from classical metal in that they are microelectrodes flexible and have much larger diam eters at the tips (32 vs. 0.1 to 1.0 ^m). This method allows us to maintain recordings from single cells even fol on lowing rather violent movements the part of the cat (Fig. 9), and therefore allows us to study the ac tivity of the same neuron over long periods of time (often several days). Prior to examining the effects of hal lucinogenic drugs on both behavior and the activity of serotonin-con taining neurons in the cat, we felt that itwas important to provide a general context for these data by first char acterizing the spontaneous activity of across the sleep these neurons continuum (27). wakefulness-arousal During a quiet waking state, sero tonergic neurons discharge with the slow, regular pattern that character izes the activity of serotonergic neu rons in anesthetized rats. However, in both this activity can be modulated directions, depending on the state of

* *T

* * *

* ** *.*.*

records the electrical Figure 9. An oscilloscope neuron activity of a single serotonin-containing in an awake, freely moving cat, such as the one

shown activity, charge.

in Fig. each

8. In this 20-second sample of dis vertical line is a neuronal

the cat. The activity increases during periods when the cat becomes active, and briefly increases still further in response to an arousing or alerting stimulus (e.g. a click or flash). On the other hand, the activity of these cells as the cat becomes quies decreases cent and drowsy (Fig. 10). Their ac tivity decreases still further when the cat enters the first phase of sleep, and finally ceases during the next stage of sleep (termed REM sleep because of of rapid eye move the appearance ments). These data, in conjunction with other evidence, recently led us to that the oft-noted propose phe similarity of dreams nomenological (which occur most vividly in REM hallucina sleep) and drug-induced in part, by tions might be mediated,
a common neurochemical event?

inactivation of central serotonergic neurotransmission (28). With the basic characterization of the activity of these neurons completed, we turned to directly examining the effects of hallucinogenic behavioral drugs, while simultaneously recording the activity of serotonin-containing neurons. We will first describe our results with 5-MeODMT (29), be cause they were the most straight forward. This drug produced dose dependent decreases in the activity of
serotonergic neurons and dose-de

pendent increases in specific behav iors (the limb flick response is the most reliable and easiest to quantify). the onset, offset, and Furthermore, effects of 5 behavioral of the peak were temporally corre MeODMT
1979 July-August 401

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

Figure 10. The activity of serotonin-containing neurons varies dramatically as a function of the state of the animal. This figure illustrates the of the same serotonin-containing activity neuron

across the complete sleep-waking Each continuum. arousal strip shows 60 sec onds of data, and each vertical line represents or spike. The highest rate of a cell discharge, is seen during active waking; the ac discharge tivity slows somewhat during quiet waking and as the animal becomes drowsy. A dramatic as the animal occurs enters sleep slowing

f:;|j||||i?^|]:;:

(slow-wave sleep-1 through slow-wave sleep-3), silent during the until the cell finally becomes of sleep, when vivid dreaming REM phase typically takes place.

llJpllllll ISHHlill

lliiilllll
in a dose of 50 Figure 11. An injection of LSD, Hg/kg of body weight, rapidly and dramatically affects the activity of serotonin-containing the cell's activity neurons. Within 15 minutes has significantly slowed, and it reaches its nadir

im
30 min

after the injec 45-60 minutes approximately de tion. This cell's activity was maximally creased by about 75% from the pre-drug base line. The cellular activity returns to normal in 4-6 hours.

45 m?n

6-hrs

lated with the onset, offset, and peak of the changes in neural activity. By directly correlating behavioral changes with changes in the activity of serotonin-containing neurons, this study provided perhaps the strongest direct support for the serotonin hy drug ac pothesis of hallucinogenic tion. One of the most interesting as pects was the finding that significant behavioral changes were often asso in raphe ciated with small decreases unit activity (e.g. 15-20 percent). This indicated that rather subtle varia tions in the outputs of these neurons might have profound behavioral ef
fects.

effects of LSD, the re physiological sults were in general very similar to but those seen with 5-MeODMT, with two important differences. First, of serotonergic duced a depression neuronal activity that lasted for ap proximately 4 hours (Fig. 11), while effects lasted for at the behavioral least 6-8 hours. Second, when the 50 the jug/kgdose was re-administered next day, it produced little or no be havioral effect, but the neuronal change was as large as that on the previous These day (30, 31).

a high dose of LSD

(50 Mg/kg) pro

is exerting its primary depres LSD sant effect on serotonin neurons, it is also producing a change in some set of postsynaptic neurons that will outlast the primary effect and continue to mediate the behavioral change. This is supported by the experiment de scribed above, inwhich we saw toler ance develop to a single dose of LSD while this dose was still exerting its primary behavioral effect. General support of these notions re in postsynaptic about changes came from ceptors experiments using in rats, in gross behavioral measures which we observed that repeated ad re of LSD markedly ministration duces the sensitivity of serotonergic (32). We have target neurons to LSD also found that repeated adminis

two

somewhat

anomalous

to ex When we used this approach and electro amine the behavioral

402 American Scientist, Volume 67

behavior outlasting findings?i.e. neuronal change and neuronal change without behavior?are probably in terrelated. It appears that even while

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

tration of LSD decreases the number of postsynaptic binding sites for both and may affect serotonin and LSD the affinity of serotonin for its re adminis ceptor sites (33). Repeated tration of LSD might therefore result in a decreased capacity of LSD and/or serotonin to stimulate neurons post neurons. synaptic to serotonergic is not med Thus, tolerance to LSD iated by a change in responsivity or sensitivity on the part of serotonergic neurons, but an important change seems to occur at the next neuron in is not clear at the series. What present, however, is the exact nature of the change in the postsynaptic
neurons.

concept is the fact that these neurons appear to be among the first to dif ferentiate during the development of is also CNS. This the mammalian supported by the fact that serotonin neurons discharge, in a wide variety of situations, with an almost clocklike rats in anesthetized regularity?e.g. (34), in awake, freelymoving cats (27), and even when examined in 400-yum thick slabs of rat brain tissue main tained in vitro (35). The slowness and regularity of the activity of these neurons probably denotes a tonic neuronal function, as opposed to a rapid and variable ac tivity, which would carry more in formation and subserve a more dy namic function. A group of farsighted who were among neuroanatomists, the first to study the raphe, specu lated that it is "probably a primitive part of the brainstem which shows relatively little differentiation during the phylogenetic ascent of the verte one would brates. Correspondingly, be inclined to ascribe to it relatively simple, but fundamental and impor tant tasks in the function of the brain" (36).

a complete explanation will, of ne cessity, involve a good deal more than the activity of one set of neurons. are also many unanswered There questions about the actions of LSD, such as the precise mechanism underlying tolerance. An important sign of health and vitality in any sci

is change and revision. Neuroscience at present one of the more vigorous fields of scientific investigation, and we therefore have no doubt that the story we have told will undergo sig and exten nificant modification
sion.

entificfield is the abilityto undergo

Why is the relationship between be so havior and neuronal change straightforward with 5-MeODMT so complex with and apparently LSD? We do not yet have the answer to this at the cellular level, but we do know that no other hallucinogenic in its capacity drug approaches LSD to produce rapidly developing, long lasting, and dramatic tolerance. This remains one of the more intriguing unanswered questions regarding of action. LSD's mechanism

References
1. A. 1968. Psychotomimetic Hofmann. the Central In Drugs Affecting agents. Nervous ed. A. Burger, pp. 184 System, 85. Marcel Dekker. Psychoses, 1954.

1968. Chemical 2. L. E. Hollister. p. 34. Charles C. Thomas. 3. J. H. Gaddum

and K. A. Hameed.

5-hydroxytryp Drugs which antagonize 9:240-48. tamine. Brit. J. Pharm. 1954. A bio and E. Shaw. 4. D. W. Wooley chemical and pharmacological suggestion PNAS disorders. certain mental about 40:228-31. 5. A. Cerletti and E. Rothlin. 1955. Role of in mental disease 5-hydroxytryptamine to lysergic acid deriv and its antagonism 176:785-86. atives. Nature and K. Fuxe. 1964. Evidence 6. A. Dahlstrom for the existence of monoamine-containing neurons in the central nervous system. I. in the cell of monoamines Demonstration bodies of brainstem neurons. Acta Physiol. Scand. 62:Suppl. 232,1-55. for the existence 7. K. Fuxe. 1965. Evidence neurons in the central ner of monoamine of vous IV. The distribution system. in the central ner monoamine terminals Scand. 64: vous system. Acta Physiol.

General

implications

including humans, do ingest plants contain the compounds, hallucinogenic ing system of serotonin neurons must have evolved to subserve some adap tive function other than mediating A large behavioral hallucinations. literature indicates that serotonin may play a general inhibitory role with respect to a variety of sensory Since animals, not commonly
motor

nin neurotransmission, whether by destruction of serotonergic neurons, inhibition of its synthesis, or blockade of its receptors, consistently produces an animal that is hypersensitive to stimuli all environmental virtually in virtually all sit and hyperactive uations. Through a general inhibitory function, serotonin neurons may an organism's be serve to modulate it within nar havior and maintain rowly specified limits.

processes.

Blockade

of seroto

It seems reasonable that some basic role would be served by neurons whose cell bodies are localized in the lower,more primitive, portions of the brain and whose axons reach widely and diffusely throughout the central nervous system. Consistent with this

Our studies of serotonin neurons indicate that as overall level ofmotor activity or arousal increases, so does the activity of these cells. Recipro cally, as the animal becomes quies cent and drowsy, the activity of these cells declines, possibly because this inhibitory control is no longer neces sitated. As the animal enters sleep, the cells fire still more slowly, and sleep, a state in which during REM tonic muscle activity is abolished, the cells stop firing. If we consider the action of hallucinogenic drugs in this context, we see a fully awake animal with a brain serotonin system func tioning as though the animal were asleep. This may provide an impor tant insight into understanding hal lucinations and perhaps, more gen erally, other altered states of con sciousness. In a given behavioral sit uation, an altered state of conscious ness may occur when a key brain such as the serotonin mechanism, system, functions in a manner that is appropriate to a different behavioral situation. We have tried to explain the behav in terms of ioral effects of LSD in of the activity single brain changes cells. As with any complex behavioral process and any centrally acting drug,

Suppl. 247, 41-85. 1961. Effects of LSD-25 8. D. X. Freedman. on brain serotonin. J. Pharm. Exp. Ther. 134:160-66.

and D. X. R. A. Lovell, 9. J. A. Rosecrans, 1967. Effects of lysergic acid Freedman. on the metabolism of brain diethylamide Biochem. Pharmac. 5-hydroxytryptamine. 16:2011-21. W. E. Foote, and M. H. 10. G. K. Aghajanian, 1968. Lysergic acid diethylamide: Sheard. neuronal units in the midbrain Sensitive 161:706-08. raphe. Science W. E. Foote, and M. H. 11. G. K. Aghajanian, of psychotogenic 1970. Action Sheard. J. on midbrain raphe neurons. drugs Pharm. 12. H. Exp. Ther. 171:178-87. 1974. and G. K. Aghajanian. J. Haigler and serotonin: Lysergic acid diethylamide of effects on serotonergic A comparison neurons and neurons receiving a seroton ergic input. J. Pharm. 688-99. Exp. Ther.

188:

and J. L. H. J. Haigler, 13. G. K. Aghajanian, 1975. Amine receptors in the CNS Bennett. In in brain. III. 5-Hydroxytryptamine

1979 July-August 403

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions

Vol. Handbook of Psychopharmacology, 6, ed. L. L. Iversen, S. D. Iversen, and S. H. Snyder. Plenum Press. 14. S. S. Mosko and B. L. Jacobs. 1977. Elec evidence against negative trophysiological from the forebrain feedback neuronal raphe unit activity. controlling mid-brain Res. 119:291-303.

21. M.

Usefulness and LSD

in studyingtheduration ofaction ofLSD

the onset and duration in the cat. Brain Res. of tolerance 132:315-26.

E. Trulson and B. of an animal

L.

Jacobs.

1977. model

behavioral

their phenomenological anism mediating & Biobehav. Rev. similarity. Neurosci. 59-69. 29. M.

2:

to

Brain

and W. C. 15. B. L. Jacobs, M. E. Trulson, Stern. 1976. An animal behavior model for and related studying the actions of LSD 194:741-43. Science hallucinogens. and W. C. 16. B. L. Jacobs, M. E. Trulson, in effects of LSD Stern. 1977. Behavioral of an animal behavior the cat: Proposal for studying the actions of halluci model nogenic drugs. Brain Res. 132:301-14. A. D. Stark, 17. B. L. Jacobs, M. E. Trulson, 1977. Comparative and G. R. Christoph. effects of hallucinogenic drugs on behavior 1: of the cat. Commun. Psychopharm. 243-54. and B. L. Jacobs. 1976. LSD 18. M. E. Trulson with serotonin deple acts synergistically studies in from behavioral tion: Evidence 4:231-34. Behau. cats. Pharm. Biochem. and C. Savage. 19. L. S. Cholden, A. Kurland, to reactions and tolerance 1955. Clinical J. Nerv. in chronic schizophrenia. LSD 122:211-21. Ment. Dis. 20. H. H. F. Fraser, A. Isbell, R. E. Belleville, 1956. Studies on and C. R. Logan. Wikler, I. (LSD-25). lysergic acid diethylamide and addicts in former morphine Effects of tolerance during chronic development 76: intoxication. Arch. Neurol. Psychiat. 468-78.

22. K. Von Hungen, S. Roberts, and D. F. Hill. at 1974. LSD as an agonist and antagonist 252: central dopamine receptors. Nature 588-89. 23. L. Pieri, M. Pieri, and W. Haefely. 1974. as an agonist of dopamine LSD receptors 252:586-88. in the striatum. Nature D. M. Kuhn, 24. G. R. Christoph, and B. L. 1977. Electrophysiological evi Jacobs. action of LSD: dence for a dopaminergic in the sub of unit activity Depression 21: stantia nigra of the rat. Life Sei. 1585-96. A. D. Stark, and B. L. Ja 25. M. E. Trulson, cobs. 1977. Comparative effects of hallu in cinogenic drugs on rotational behavior rats with unilateral 6-hydroxydopamine 44:113-19. lesions. Eur. J. Pharm.

E. Trulson and B. L. Jacobs. 1979. Ef fects of 5-methoxy-N,N-dimethyltrypta mine on behavior and raphe unit activity 54: in freely-moving cats. Eur. J. Pharm. 43-50.

and B. L. Jacobs. In press. 30. M. E. Trulson between the effects of LSD Dissociations on behavior in and raphe unit activity freely moving cats. Science. 31. M. E. Trulson, C. A. Ross, and B. L. Jacobs. 1977. Lack of tolerance to the depression of raphe unit activity by lysergic acid di 16:771-74. ethylamide. Neuropharm. 32. M. E. Trulson, C. A. Ross, and B. L. Jacobs. for the stimu evidence 1976. Behavioral serotonin receptors by high lation of CNS 2: Comm. doses of LSD. Psychopharm. 149-64. In press. and B. L. Jacobs. 33. M. E. Trulson of serotonin and LSD Alterations receptor binding following repeated administration

26. M.

in B. Bowers. 1972. Acute psychosis duced by psychotomimetic drug abuse. 27:437-40. Arch. Gen. Psychiat. 27. M. E. Trulson and B. L. Jacobs. 1979. unit activity in freely moving cats: Raphe with Correlation level of behavioral arousal. Brain Res. 163:135-50. This con the results of a pre firmed and extended vious study by D. J. McGinty and R. M. 1976, Dorsal Harper, raphe neurons: De of firing during in cats, pression sleep Brain Res. 101:569-75. 1978. Dreams 28. B. L. Jacobs. and nations: A common neurochemical halluci mech

ofLSD. Life Sei.

1974. Mid and B. L. Jacobs. 34. S. S. Mosko brain raphe neurons: Spontaneous activity Behau. to light. Physiol. and response 13:589-93. 1976. Re and B. L. Jacobs. 35. S. S. Mosko in raphe unit activity cording of dorsal Lett. 2:195-200. vitro. Neurosci. 1961. 36. E. Taber, A. Brodai, and P. Walberg. in the The raphe nuclei of the brainstem cat. I. Normal topography and cyt?archi J. Comp. tecture and general discussion. 116:161-88. Neurol.

"Whatever

happened

to elegant

solutions?"

404

American

Scientist,

Volume

67

This content downloaded from 134.117.10.200 on Wed, 29 Jan 2014 06:32:04 AM All use subject to JSTOR Terms and Conditions