Christopher Bennett

Amines and Affect Research Proposal

Dr. Brian Knutson
23 March 2007
 Exploring Altered Perception: The Affect of Serotonin and
Synesthesia in Hallucination

Section I: Past Research and Directions

Introduction: The Big Questions

Many experiments of the 21st century thus far have posed questions that were
earlier the territory of philosophy. Questions like: “How do we think? What is
consciousness? And how and what are we really perceiving?” have introduced us to new
and exciting grounds for exploring the brain. The fundamental problem in these
experiments thus far has been a disconnect between the fundamentally subjective
experience of the mind and the complex biochemical systems of the physical brain. With
new fMRI, radioligand, and electrophysiological studies, however, we have come closer
to correlation than ever, as one can compare introspective evaluations to actual brain
activity. An interesting question is how the experience of the mind can be grasped by
measuring alterations upon it, or by altering the biological systems of perception. Altered
perception is found in two well-known and recently biologically traced systems:
psychoactive hallucination, and synesthesia. Before posing an experimental framework to
test perception, however, we need to fully understand the nature and implications of these
phenomena.
Serotonin Systems and Hallucinogens
Hallucination is an extremely powerful phenomena, and extremely complex; a
hallucination can induce drastic changes in mood, perception, cognition, and mental
processes. Yet hallucination has been consistently traced to one biogenic amine:
serotonin, or 5HT. LSD, or Lysergic Acid Diethylamide, is the most well known and
most heavily studied of the sertonergic hallucinogens, although it belongs to a broader
group of chemicals which have the same pharmacological effect including the
phenylisopropylamines, such as DOB, DOI (4-iodo-2,5-
dimethoxyphenylisopropylamine), and the indolealkylamines, such as DMT or
dimethyltryptamine. Generally, these chemicals are all thought to act as 5HT receptor 2
agonists, meaning that they directly depress 5HT containing neurons at the 2A receptor
sites, changing feed-back mechanisms and agnozing serotonin release primarily in raphe
neurons. Since LSD is heavily involved in 5-HT terminal fields throughout its activity
and later antagonizes raphe neurons, this model is obviously simplistic. In recent studies,
George Aghajanian has proposed that LSD must mostly or entirely effect raphe neurons
through the 2A receptor mechanism, causing decreased response but increased 5HT
activity. (Cooper 300). However, there are still problems with this idea; raphe-lesioned
animals still experience the behavioral effects of LSD, and additional 5HT seems to
increase rather than decrease response. Clearly, the exact mechanism of LSD remains a
cellular “jigsaw puzzle” (301).
Recent Studies: Clarifying the Affect of Hallucination
 Although the mechanisms of causation are extremely complicated, there is an
undisputed correlation between the administration of a 5HT 2A agonist and hallucination.
In a study by Sadzot, et al. entitled “Hallucinogenic drug interactions at human brain 5-
HT2 receptors: implications for treating LSD-induced hallucinogenesis” the authors seek
to establish conclusively the implication of the 5-HT 2A mechanism. Using radioligand
binding techniques, they used human cortical tissues in order to determine the binding
patterns of 13 psychoactive substances, including all of the serotonergic hallucinogens.
LSD was the most potent ligand displacer, followed by DOI, DOB, and DMT to a lesser
degree. Further, the authors found an almost one-to-one correlation (r= 0.97) between
hallucinogenic potencies and affinities, meaning that the 2A system was almost
universally involved in hallucinogenic action. In addition, the glutamate mechanism has
recently shed light on how serotonin may influence behavior above and beyond 5HT
innervation; Glutamate is involved in other psychoactive processes like PCP and
influences the 5HT 2A mechanism. A recent study by Lambe, Aghajanian, et al. entitled
“Hallucinogen-induced UP states in the brain slice of rat prefrontal cortex: role of
glutamate spillover and NR2B-NMDA receptors” has attempted to connect the role of
glutamate spillover in NDMA pathways of the brain and the changes in cognition
producing the affect of hallucination. Electrophysiological studies in the rat prefrontal
slice have shown that both LSD and DOI enhance a prolonged, late wave of glutamate
release after an electrical stimulus, however in this study the authors attempted to trace
the initial 5HT 2A action to a late excitatory postsynaptic current (EPSC) using
microdialysis in rats. The EPSC results in a phasic glutamate spillover that alters several
sensory and cognitive pathways, which may explain part of the affect of hallucination.
Recent studies have also attempted to explain the affect of hallucination by comparing
the psychological effect of hallucinogens to that of mental illness, especially
schizophrenia. A recent study, “Psychological effects of (S)-ketamine and N,N-
dimethyltryptamine (DMT): a double-blind, cross-over study in healthy volunteers” by
Heekeren, et al, compares the positive and negative affect of hallucination to the
symptoms of hallucination. The authors administered (S)-ketamine, a glutamine
hallucinogen, and DMT, a serotonergic hallucinogen, and measured the differential affect
in double-blind. Their results attempted to compare the affects of the psychoactive drugs
to the endogenous conditions correlated with psychosis, especially in schizophrenia; they
found that DMT correlated to the positive or hallucination and thought disorder of
psychoses, while (S)-ketamine correlated to the negative or catatonic effects. The authors
concluded that the 5-HT 2A and glutamine systems stimulated by psychoactive drugs
may be naturally occurring models of altered cognition. These studies seem to imply that
regardless of pathway, serotonin is intimately involved in hallucination, creating and
mimicking brain conditions that significantly change the nature of cognition.
Synesthesia: An Introduction
Synesthesia, most broadly defined, is the coupling of sensory pathways. It is a
relatively rare hereditary condition. In the past, synesthetes were either declared liars or
loopy because of skepticism; even after Synesthesia was confirmed as an actual
perceptual and cognitive condition, it has been treated as a curiosity of the brain and no
more until very recently. Synesthesia is not just a natural brain condition; it also often
experienced by users of psychoactive drugs and occasionally the mentally ill. Synesthesia
is now being viewed as an important and interesting window into the workings of the
mind, especially the nature of cognition, perception and language. There are several types
of synesthesia: grapheme-color synesthesia, where synesthetes view letters and numbers
as possessing innate colors, music-color synesthesia, where viewers experience patterns
and colors while listening to music, and rarer types where individuals might experience
tactile properties or personalities with letters or numbers or tactile properties when tasting
different foods. The most widely studied and first confirmed synesthesia was grapheme-
color synesthesia. A great deal of research has confirmed that synesthesia is automatic,
perceptual rather than cognitive, and automatic rather than voluntary. A “Synesthesia
stroop test,” has been developed and tested many times, in which synesthetes pick out a
certain number out of a field of other numbers. The fact that synesthetes consistently pick
these numbers out of a field faster than the control time validates that these colorful
numbers would pop out to them. Two important scholars in the field are EM Hubbard and
VS Ramachandran. Their 2001 paper “Synesthesia: A Window into Thought, Perception
and Language” was a step forward in advancing the importance and influence of research
in synesthesia. In addition to collating older research, Hubbard and Ramachandran
proposed cross-activation theories, explored the top-down and involuntary nature of
synesthesia, and explored the role of synesthesia in the evolution of language and the
problem of qualia.

Figure 1: A “Synesthesia Stroop Test” (Hubbard, et al)

Synesthesia: Studies in Cross-Wiring and Brain Activation
With new experimental methods in the past few years, however, analysis of
synesthesia has become less psychological and more biological, leading to new and
interesting implications for the importance of synesthesia in looking at perception. New
brain imaging techniques, specifically fMRI, have played a fascinating role in confirming
the role and substrate of synesthesia. Aleman, et al investigated and confirmed the nature
of cross-wiring and brain facilitation in a study entitled “Activation of striate cortex in the
absence of visual stimulation: an fMRI study of synesthesia.” This study focused on a
color-word synesthesia in a single subject with synesthesia, but tested it in a more
rigorous way; by blindfolding the subject and simply reading her words, they tested
whether the subject experienced independent visual stimulation in the striate cortex that
correlated to the ‘color’ of the word read to her. The study revealed a direct stimulation
of this area of the cortex, and an amazing consistency on the type of color stimulated and
the consistency of that over time. This study and others like it have provided undeniable
evidence of synesthesia, as well as evidence of local cross-activation of sensory neural
components. In “The Neurocognitive mechanisms of synesthesia,” Hubbard and
Ramachandran provide a more thorough and neural basis for synesthesia using recent
studies. Expounding on earlier theories of “cross-activation”, or neural coupling between
the different cortexes, they explore two different theories for the neural cause: dis-
inhibition or a local cross-activation from a lack of brain pruning. The first theory
stresses disinhibited transmission in a “supersensory nexus” (such as the temporo-
parietal-occipital junction) which regulates sensory signals differently in those with
synesthesia, while the second stresses a local cross-wiring of sensory pathways, probably
originating in a lack of brain differentiation or pruning in these areas. They argue these
mechanisms are probably both in place to some degree, although the cross-activation
hypothesis has been more easily verified by fMRI studies showing local connections such
as in Aleman, et al. Finally, they pose the question of whether or not the synesthetic
pathways exist in non-synesthetes. They stress that the answer must be yes, even if
pruning is mostly true; there seem to be fundamental metaphoric connections between
senses in language and certain sensory combinations (shapes to sounds, pitches to light),
as well as voluntarily experienced synesthesia, such as psychoactive drug use.

Figure 2: A ventral fMRI view of the brain. HV4 regions (tied to color) are activated
clearly (independently) only in the synesthetes. (Hubbard, et al)
So What Causes Altered Perception? The Experimental Question
Altered perception is still a mystery. However, the best way to take a step forward
is by making good questions that may produce interesting experimental results. Insofar
as hallucination fundamentally alters how our perception operates, and synesthesia
represents a curious coupling of the senses that form perception, both of these
abnormalities may shed light on the pathway and nature of perception itself. It seems
best, then, to investigate the common denominators behind both phenomena. A central
question seems to be: is there a common structural or biochemical basis behind the
altered sensory perceptions of synesthesia and hallucination? It seems that if synesthesia
can be experienced by non-synesthetes taking psychoactive drugs, there are two
overlapping possibilities: either the structural changes that produce the experience of
synesthesia must be mimicked, or that a biochemical situation is being produced in the
brain that occurs in synesthetes as well. I would like to test these two possibilities under
the influence of a psychoactive drug, testing the relationships between serotonin in the
brain in relation to hallucinogens and possibly synesthesia, and the cross-activation
behind synesthesia and possibly involved in hallucination itself. In short, the
experimental question seems to be whether serotonin and cross-activation are
independent (mimicked) or interrelated systems that produce altered perceptions.
Section II: An Experimental Proposal
Experimental Design and Procedure
This study will involve a double-blind administration of DMT and a placebo with
a group of 20 healthy volunteers (Age Range: 20-30) with some former use of
psychoactive drugs. The first independent variable is that the pool will divided for
purposes of cross-activation; 10 participants will be synesthetes, 10 will be non-
synesthetes. the pool will be approximately half male and half female, and the
synesthetes will be evenly split between left-handed and right-handed individuals. Of
each group (S, NS) half will be administered the DMT and half will be administered the
placebo. I selected DMT as the second Independent Variable because it as a reliable 5HT
2A agonist, and although not as potent as LSD, it will a safe and predictable (like in
Heekeren, et al). Synesthetes will all be selected as grapheme-color synesthetes, and they
both supposed S and NS will be screened with a Synesthesia stroop test beforehand to
ensure accurate pools. This experiment will seek to collect all the data needed in a 2-day
experimental period; for statistical purposes, I want to collect two sets of independent
data. Both days will involve the administration of the placebo and DMT to the groups by
an unbiased administrator. During the course of the experiment, I will seek to monitor
three dependent variables. The first will be an analysis of the participant’s synesthetic
experiences. Every 30 minutes after the first 30 minutes, the patients will be verbally
asked if they are experiencing several phenomena: dizziness, confusion, hallucination,
strange shapes, and several synesthesia-like experiences. Those who report any type of
phenomena (both those on placebo and those not, both NS and S) will be tested in the
Stroop format. Here, we will be attempting to see if any S participants lose their
experience of synesthesia, or if any NS participants have verified synesthesia. The
second variable will be an fMRI analysis in order to screen for potential cross-wiring of
the patients on DMT, both the NS and S in the experiment. These fMRIs will be
administered 3 times during the experiment to both those on DMT and the placebo, at
t=30, 60, and 120 minutes. The third variable will be measurement of 5HT activity. Since
it is difficult to measure serotonin levels endogenously in the brain, we will use Positron
Emission Tomography method or PET. By marking serotonin as an isotope, we should be
able to measure and look at the levels and placement of 5HT throughout the brain. After
injecting all patients with the isotope an hour before the drug is administered, we will
measure two times, at t=15 minutes before significant changes, and a second time at t=90
minutes at the height of the trip. We believe this will provide a vast amount of controlled
and accurate data on synesthetic and sertonergic activity in the brain during the
psychoactive trip.
Hypothesis/ Predicted Results
We predict that there is little if any interrelation between the systems of
serotonergic hallucination and synesthesia; we believe that hallucination itself is simply
mimicking certain features of synesthesia through the massive changes it has on
biochemical pathways, and the consistent experience of synesthesia is largely structural.
As far as the predicted data, we believe that we will in fact have several NS participants
who will have confirmed synesthetic experiences, both by the Stroop test as well as
cross-activation in the fMRI. We believe it is unlikely, however, that S participants will
not experience synesthesia, or that they will lack cross-activation in their fMRI scans. We
believe that almost all patients on the hallucinogen will report substantial sensory and
cognitive changes, and that will be correlated with high activity in the visual cortex, even
if there is no cross-activation. Finally, we believe that PET scanning will not reveal
anything especially significant. Although 5HT levels will vary from individual to
individual, we believe they will significantly rise in all persons given DMT between the
two measurements, especially in the raphe areas of the serotonergic pathways, and
obviously be lower at times for the control group. However, we do not believe there will
be a significant difference between the S and NS groups in 5HT levels or distribution in
the brain. If a consistent difference in 5HT patterning exists, we believe it will exist
between the NS individuals on DMT who did experience synesthesia versus those who
did not. This may reveal something about the chemical pathways, either in the visual
cortex, occipital-temporal junction, or other areas, that give rise to the temporary cross-
activation we hope to confirm in fMRI.
Confounds and Complications
The most obvious confound is the idea of individual response to drugs, especially
given the small sample size. It is possible that many individuals on DMT will not have
significant hallucinogenic experience. In a much worse scenario, it is possible that no NS
participants will experience synesthetic phenomena. Another and more dangerous
confound is that other neuronal systems or biochemical pathways are heavily or actively
involved in the hallucination process, even more than serotonin. If this were true, any
conclusions we can draw would be speculatively correlational and not causative at all,
since serotonin levels would not be uniquely correlated to the hallucinogenic state as we
assumed in the experiment. A final confound is that synesthesia may be improperly
measured by fMRI, or possibly exist and go undetected by fMRI. It is often difficult to
confirm borderline cases of synesthesia by fMRI since one must establish a significant
difference in cross-activation, and it is possible that hallucination-induced synesthesia
may fail to be confirmed at all.
Future Directions and Experiments
If the hypothesis is too pessimistic and deeper connections emerge, the next step
is to investigate this more thoroughly; experiments could include measuring and tracking
the structure of naturally occurring serotonin systems in synesthetes, or during more in-
depth fMRI or PET studies on those NS who experienced synesthesia during
hallucination. More likely, however, the next step involves looking at the chemical
pathways that were altered in those NS who experienced synesthesia. This may be of
interest in understanding more of the hallucination jigsaw puzzle, as well as in
understanding how this ‘capacity’ for synesthesia was triggered in terms of the pathways
of glutamate, serotonin, and other neurotransmitters systems we did not analyze. And
even though we predict that synesthesia is largely structural (due to cross-wiring), we
wonder if the ‘hybrid’ cases in our experiment could spur us to experimentally explore
the ‘marginal’ synesthesia we all can recognize in metaphors, and use this to better
understand the systems of language, consciousness, and perception.

Works Cited

Aleman A, Rutten GJ, Sitskoorn MM, Dautzenberg G, Ramsey NF.

“Activation of striate cortex in the absence of visual stimulation: an fMRI study of
synesthesia.” Neuroreport. 2001 Sep 17;12(13):2827-30.

Cooper, Jack, Bloom, Floyd, and Roth, Robert. The Biochemical Basis of Pharmacology.
Oxford: Oxford University Press, 2003.

Heekeren K, Neukirch A, Stoll M, Stock C, Obradovic M, Kovar KA.

“Psychological effects of (S)-ketamine and N,N-dimethyltryptamine (DMT): a double-
blind, cross-over study in healthy volunteers.” Pharmacopsychiatry 2005; 38: 301-311

Hubbard, EM, and Ramachandran, V.S. “Synaesthesia- A Window Into Perception,

Thought, and Language.” Journal of Consciousness Studies, 8, No. 12, 2001:3-34.

Hubbard EM, Ramachandran VS. “The Neurocognitive mechanisms of synesthesia.”

Neuron. 2005 Nov 3;48(3):509-20.

Lambe, Evelyn K. Aghajanian, George K. “Hallucinogen-induced UP states in the brain

slice of rat prefrontal cortex: role of glutamate spillover and NR2B-NMDA receptors.”
Neuropsychopharmacology. 31(8):1682-9, 2006 Aug.

Sadzot B, Baraban JM, Glennon RA, Lyon RA, Leonhardt S, Jan CR, Titeler M.
“Hallucinogenic drug interactions at human brain 5-HT2 receptors: implications for
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