PDFlib PLOP: PDF Linearization, Optimization, Protection

Page inserted by evaluation version

www.pdflib.com sales@pdflib.com
Endogenous b-Carbolines as Clonidine-
Displacing Substances
E.S.J. ROBINSON,a N.J. ANDERSON,a J. CROSBY,b D.J. NUTT,a
AND A.L. HUDSONa
aPsychopharmacology Unit, School of Medical Sciences, University Walk,
Bristol BS8 1TD, UK
bSchool of Chemistry, School of Medical Sciences, Bristol BS8 1TD, UK

ABSTRACT: Endogenous b-carbolines, such as harmane, are known to occur in

mammalian species including humans. Radioligand binding studies have re-
vealed that certain b-carbolines display high affinity for both I1 and I2 imidazo-
line-binding sites (IBS). Functional studies have shown that the b-carboline
harmane elicits many characteristics expected of an endogenous ligand IBS. This
article discusses the evidence relating to b-carbolines as endogenous ligands and
presents a case for harmane and related compounds as endogenous ligands
for IBS.
KEYWORDS: b-carboline; clonidine-displacing substance (CDS); harmane

INTRODUCTION

In 1999, Hudson and coworkers1 reported that certain -carbolines display high
affinity for imidazoline-binding sites (IBS). In a subsequent publication, Husbands
et al.2 also reported that a number of -carbolines display high affinity for I1- and
I2-binding sites. It was particularly interesting that some of the -carbolines that
were shown to have high affinity also occur endogenously in mammalian tissue,3
raising the possibility that harmane, norharmane, and tetrahydro-norharmane (THN)
may represent endogenous ligands for IBS.
Since the discovery of the IBS, a number of candidates have been put forward as
endogenous clonidine-displacing substances (CDS). The first studies of CDS de-
scribed an impure extract isolated from calf brain.4 The fact that this extract was able
to displace [3H]clonidine fully from rat brain membranes suggested the extract con-
tained an endogenous ligand for IBS.4 Since then, a number of different candidates
have been put forward as CDSs. The first substance identified as a CDS was agma-
tine, which was extracted using a modified CDS-extraction procedure.5 Agmatine,
however, does not meet many of the criteria for classical CDS and is unlikely to rep-
resent the active component of the original extract. Further studies have since iden-
tified imidazoleacetic acid-ribotide as a CDS, but, again, this substance is not
believed to represent the active component of classical CDS.6 Research in our labo-

Address for correspondence: A.L. Hudson, Psychopharmacology Unit, School of Medical Sci-
ences, University Walk, Bristol BS8 ITD, UK. Voice: (+44) 117 928 8608; fax: (+44) 117 928 9700.
e-mail: a.l.hudson@bristol.ac.uk
Ann. N.Y. Acad. Sci. 1009: 157166 (2003). 2003 New York Academy of Sciences.
doi: 10.1196/annals.1304.018

157
158 ANNALS NEW YORK ACADEMY OF SCIENCES

ratory has recently identified -carbolines within the active fraction of bovine lung CDS
(J. Crosby. 2003. Harmane and harmalan are active constituents of bovine lung CDS
[unpublished report]). This paper discusses recent findings in relation to identifica-
tion of the active component of classical CDS and the evidence supporting the -car-
bolines as endogenous ligands at IBS.

CLASSICAL CDS

Atlas and Burnstein4,7 undertook the first experiments to ascertain whether there
was an endogenous compound that had clonidine-like activity. A methanolic extrac-
tion of rat7 and bovine4 brain, followed by reverse-phase high performance liquid
chromatography (RP-HPLC) purification yielded a partially purified extract exhib-
iting such activity. This substance, termed classical CDS, was divided into the crude
and the RP-HPLCpure forms.
Research in our laboratory has been aimed at identifying the structure of the
active component of classical CDS. Using the method described by Singh and co-
workers,8 a crude methanolic extract from bovine lung was prepared, and, when the
preparation was run on a reverse-phase C18 column with a methanolic gradient, a
three-peak profile as described by Atlas and Burnstein was demonstrated.4 Using ra-
dioligand binding studies, the elution time of the active fraction was ascertained, and
electrospray mass spectroscopy was used to determine mass fragmentation patterns
and structural properties. Using this technique, tryptophan was identified as an inac-
tive contaminant of CDS.9 Subsequent studies also identified an inactive -carboline,
1-carboxyl tetrahydro--carboline. These substances made it difficult to detect and
identify other molecules that were eluting in the active fraction but at significantly
lower levels. A number of potential candidates were run on HPLC, and a -carbo-
line, harmalan was shown to elute within the active fraction. The extraction proce-
dure was subsequently revised and optimized for the detection of -carbolines. Both
harmane and harmalan were shown to be present in bovine lung.

ENDOGENOUS b-CARBOLINES

The nomenclature used for -carbolines can be confusing. The original chemical
names, originating from 9H-pyrido [3,4-b] indoles, have been replaced by the -
carboline terminology. In this report the harmalan nomenclature and numbering of
carbon and nitrogen atoms is used as illustrated in FIGURE 1.
There is no doubt that the beta-carbolines, harmane, harmalan, tetrahydroharmane
(THN), and norharmane, display high affinity for IBS, but the question still remains
as to whether they are truly endogenous substances. Furthermore, there would need
to be some level of regulation present if these substances were involved in a neuro-
modulatory or hormonal role. Detailed studies quantifying endogenous -carbolines
have shown regional differences in their distribution as well as changes in their lev-
els under different circumstances. Their endogenous formation, however, has been
a subject for debate.
The first endogenous -carboline, pinoline, (6-methoxy-tetrahydro--carboline),
was identified from pineal gland tissue by McIsaac in 1961.10 Since its discovery
ROBINSON et al.: ENDOGENOUS b-CARBOLINES 159

FIGURE 1. Structure of norharmane (-carboline) and

standard numbering of carbon and nitrogen atoms. Har-
mane has a methyl substitution at the 1 position, which is
known to increase binding to MAO-A, whereas substitutions
at the 5, 6, and 7 position are associated with I2 binding.

various extraction, detection, and quantification methods have been used to char-
acterize these substances in biological samples (TABLE 1). Unfortunately, the study
of -carbolines has been complicated because they can be readily formed artifac-
tually. Bosin and coworkers20 found that a significant amount of artifact -carboline,
6-methoxy-tetrahydroharmane, could be formed from the condensation of endog-
enous serotonin (5-HT) with exogenous formaldehyde in the dichloromethane
organic solvent. Bosin also showed that many research groups were not using
aldehyde-trapping reagents, such as semicarbazide, or ensuring that the organic
solvents used were aldehyde-free.20 This demonstration prompted a review of the
procedures to minimize artifactual -carboline formation and demonstrate that
the compounds identified were truly endogenous substances and not artifacts of
the extraction procedure. To date, endogenous levels of harmane, harmalan, nor-
harmane, and several tetra-hydro-carbolines have been quantified in mammali-
an tissues (TABLE 1).

TABLE 1. Identification of endogenous b-carbolines in mammalian tissue

-Carboline Tissue Detection Method Ref.
Harmane Rat arcuate nucleus Spectral studies and TLC 11
Human platelets TLC, UV, fluorimetry, GC-MS 12
C18 RP-HPLC and fluorometric 13
detection
Norharmane Human plasma HPLC with fluorometric 14
detection
Rat plasma, brain, liver, kidney, 15
spleen, heart and lung
THN Rat forebrain TLC and GC-MS 16
Rat brain GC-MS 17
Human platelets GC-MS 3
6-MeO-THN Rat arcuate nucleus Spectral studies and TLC 18
Rat brain GC-MS 17
Rat adrenal gland
Pineal Gland GC-MS 19
Abbreviations: TLC, thin-layer chromatography; GC-MS, gas chromatrography-mass spec-
troscopy; HPLC, high-performance liquid chromatography; RP, reverse phase; UV, ultraviolet.
160 ANNALS NEW YORK ACADEMY OF SCIENCES

IN VIVO SYNTHESIS OF b-CARBOLINES

Evidence from in vitro studies has shown that -carbolines are formed spontane-
ously through the Pictet-Spengler cyclization of indolamines with carbonyl-containing
compounds such as formaldehyde.21 Therefore, stimulation of pathways in vivo that
lead to the formation of formaldehyde and acetaldehyde could result in increased
synthesis of -carbolines.22 An alternative route for synthesis, using glyoxylic acid,
results in the rapid formation of -carbolines from tryptamine.3 Pyruvic acid is
closely related to glyoxylic acid and occurs in mammalian tissues. In the presence
of a monoamine oxidase inhibitor, [3H]typtamine and pyruvic acid were shown to
form -carbolines in vivo (FIG. 2). This process seemed to be enzymatic, because it
did not occur in vitro under pseudophysiologic conditions.23,24 Taken together, these
data showed that some of the -carbolines of high affinity for IBS exist endogenously
in mammalian tissues. Formation of these substances may be enzymatic, although
the mechanisms have yet to be elucidated.

BINDING AND SIGNALLING

The pharmacology of -carbolines is complex because they are known to bind to

many different sites within mammalian tissue (TABLE 2). Studies investigating the
existence of an endogenous ligand for the benzodiazepine site identified -carboline
3-carboxylic acid ethyl ester (-CCE), a substance later shown to be an artifact of

FIGURE 2. Proposed synthetic pathway for endogenous -carbolines from tryptamine

and pyruvic acid through the intermediate products 1-carboxyl-tetrahydroharmane (1-CTHH)
and harmalan.
ROBINSON et al.: ENDOGENOUS b-CARBOLINES 161

TABLE 2. Examples of endogenous b-carboline binding sites

Binding Site Compound and Affinity Ref.
Benzodiazepine site Harmane and norharmane (MA) 25, 26
MAO-A Harmane (HA), norharmane (MA) 27
MAO-B Norharmane (HA), harmane (MA) 27
Monoamine re-uptake
sites THN (MA), Pinoline (MA) 28, 29
G-proteins Harmane, norharmane and THN (MA) 30, 31
Cytochrome P-450 Norharmane (HA) 32
DNA intercalation Harmol>harmine>harmaline>harmane>norharmane 33
Unknown Norharmane (HA) 34
Abbreviations: HA, high affinity (<100 nM); MA, moderate affinity (100 nM100 M).

the extraction procedure. The endogenous -carbolines have moderate affinity at the
benzodiazepine site, but they show interaction with aspects of monoamine neuro-
transmission. Harmane and other -carbolines with a methyl substitution in the 1 posi-
tion are inhibitors of monoamine oxidase (MAO) A.27 In addition, affinity for the 5-HT
reuptake site has been reported for THN and pinoline.28,29 It is unlikely that concentra-
tions of endogenous -carbolines are high enough for these targets to represent their pri-
mary sites of action. Furthermore, radioligand-binding studies have suggested that
harmane and norharmane bind to an unknown high-affinity site in the brain34 and adre-
nal glands.27 In 1997, Lichtenberg-Kraag and coworkers30 showed that harmane and
norharmane could activate small G-proteins in a human neuroblastoma cell line and that
this effect was dose-dependent with a bell-shaped dose-response curve. The SHSY5Y
cell expressed a high-affinity [3H]norharmane site, and it has been concluded that acti-
vation of this -carboline receptor facilitates phosphoinositide-specific phospholipase
C activation by the muscarinic agonist carbachol.30 In a different study, norharmane
and harmane were shown to activate G-proteins in a receptor-independent manner with
EC50 values of 60 M and 300 M, respectively.31 Unfortunately, no further studies
into the signaling mechanisms of endogenous -carbolines have been reported.

FUNCTIONAL CHARACTERISTICS OF IBS AND

ENDOGENOUS b-CARBOLINES

The -carbolines have been shown to induce a diverse range of functional effects
often attributed to the inhibition of MAO-A. The -carbolines are selective inhibi-
tors of MAO-A and, with the exception of norharmane, are only weak inhibitors of
MAO-B.27 As a consequence of their interactions with the MAOs, many of the function-
al properties associated with endogenous -carbolines can be attributed to modulation of
noradrenaline, dopamine, and 5-HT. Many of the -carbolines found in plants are also
potent monoamine receptor ligands.35 Their binding to 5-HT2A receptors is associated
with hallucinations.35 Among the brain -carbolines, only pinoline is believed to be hal-
lucinogenic, perhaps because of its high affinity for the 5-HT re-uptake site.30
162 ANNALS NEW YORK ACADEMY OF SCIENCES

TABLE 3. Summary of functional data for endogenous b-carbolines and IBS

Ref.
Harmane, norharmane, 1-methyl harmane and tetrahydro--carboline
displace [3H]2-BFI and/or [ 3H]clonidine with low nanomolar affinity. 1
IBS ligand, 2-BFI and harmane interact in vivo to modulate body
temperature. FIG. 3
Harmane induces a hypotensive response when injected into the
rostrolateral ventrolateral medulla of rats, an effect which is antagonized
by the mixed I12AR antagonist efaroxan. 36
Harmane, norharmane, and harmaline substitute for 2-BFI in the rat drug
discrimination paradigm. 37
Harmane and imidazoline ligands modulate monoamine levels in specific
brain areas. 38, 39
Norharmane and the selective IBS ligand, BU224, reduce parameters
associated with morphine withdrawal syndrome. 39, 40
Harmane is found in high density in the arcuate nucleus, a region of the
hypothalamus associated with IBS. 11, 41
Harmane and norharmane bind to a high (nanomolar)-affinity site in the
brain, which is not MAO. 34
Harmane and atypical I3 ligands modulate insulin secretion. 44
Harmane and THN modulate food intake. 45, 46

Several studies have investigated the possibility that harmane is an endogenous

ligand for IBS, and many aspects of harmane, norharmane, and THN pharmacology
overlap with that describing IBS ligands (TABLE 3). Harmane has been shown to
induce a hypotensive effect in spontaneously hypertensive rats following central ad-
ministration.36 This hypotensive effect was fully reversed by the mixed I1/2-
adrenoceptor antagonist, efaroxan. In this study harmane was equipotent with the
positive control clonidine but lacked the tachycardiac effects seen with the mixed
I1-/2-adrenoceptor agonist.36 Previous studies have shown that certain -carbo-
lines effect cardiovascular parameters,42 but this study was the first demonstration
of direct antagonism of a -carbolinemediated effect by an imidazoline ligand.
Studies in our laboratory have investigated the ability of an I2-selective ligand,
2-BFI, to antagonize the hypothermic response induced by harmane (FIG. 3). Previ-
ous studies had shown that harmane induces a hypothermic response, which is not
mediated by inhibition of MAO.38 Results from our laboratory have shown that
2-BFI pretreatment can significantly attenuate the hypothermic response induced by
harmane (10 mg kg1). At present, the mechanisms underlying this response are not
clear but may relate to an interaction with monoaminergic systems. Studies using
other I2-selective ligands and a number of different -carbolines have shown that
both classes of compounds can modulate monoamine levels in vivo.38,39
Other functional effects mediated by IBS include modulation of food intake and
regulation of insulin secretion.43 The I3 site is associated with potentiation of insulin
secretion, and harmane has been shown to induce insulin secretion.44 Furthermore,
ROBINSON et al.: ENDOGENOUS b-CARBOLINES 163

FIGURE 3. Effect of 2-BFI (10mg kg1) pretreatment on harmane- (10mg kg1)induced

hypothermia, 15 minutes after Harmane administration. Data shown as mean SE mean for
910 animals per group; statistical comparison made using a one-way ANOVA with Tukeys
test post hoc.

when given to rats, harmane and THN inhibit food intake in a dose-dependent man-
ner.45,46 These results suggest a role for endogenous -carbolines in regulating food
intake and energy homeostasis.
The -carbolines have also been extensively studied in relation to their mutagen-
esis and toxicity.47,48 Mutagenic and comutagenic properties have been reported for
harmane and norharmane. The mutagenic and comutagenic properties of harmane,
norharmane, and related compounds are associated with their interaction with DNA
and enzymatic systems such as cytochrome P-450.48 The -carbolines are also likely
to modify and increase genotoxic and toxic consequences of other compounds.48 In
relation to neurotoxicity, the -carbolinium cations formed enzymatically from har-
mane and norharmane are of particular interest. -carboline N-methyltransferase is
found in mammalian brain and catalyses the 2N-methylation of simple -carbolines.
The resulting products, 2N-methylated -carbolinium cations, are structural and func-
tional analogues of the neurotoxic agent MPP+. As such, these compounds have
been investigated as factors involved in the etiology of Parkinsons disease.47 In a
recent study, significant increases in -carboline 9N-methyltransferase activity
were observed in postmortem examinations of the frontal cortex of Parkinsons
disease patients.49 The mechanism by which these neurotoxic agents are associated
with the pathogenesis of Parkinsons disease is not known, but interaction between
N-methylated -carbolines and specific cellular proteins has been reported.50 Further
investigations will determine the significance of these findings.

DISCUSSION

Two endogenous -carbolines, harmane and harmalan, are active constituents of

bovine lung CDS and probably represent the active component of classical CDS
164 ANNALS NEW YORK ACADEMY OF SCIENCES

described by Atlas and Burnstein.4 The procedure for preparing classical CDS
involved the heating of tissue and was likely to result in the further production of
-carbolines by spontaneous formation. Therefore the original extraction protocols may
have resulted in high levels of artifactual -carbolines. Future studies of CDS should
strive to use extraction protocols that prevent artifactual formation of -carbolines.
Several different research groups have shown that a number of different -carbolines
can be extracted from mammalian tissues and that these -carbolines represent en-
dogenous molecules. Furthermore, mechanisms for their endogenous synthesis,
source, and site of action have been hypothesized. Taken together, these substances
and the IBS represent a novel neuroendocrine system. Future studies of -carbolines
and IBS in combination should provide greater insight into the existence and impor-
tance of this proposed system.

ACKNOWLEDGMENTS

This work was funded by the Wellcome Trust and Roche Biosciences, Palo Alto,
CA, USA.

REFERENCES

1. HUDSON, A.L., et al. 1999. Harmane, norharmane and tetrahydro--carboline have

high affinity for rat imidazoline binding sites. Br. J. Pharmacol. 126: 2P.
2. HUSBANDS, S.M. 2001. -Carboline binding to imidazoline receptors. Drug Alcohol
Depend. 64: 203208.
3. AIRAKSINEN, M.M. & I. KARI. 1981. -Carbolines, psychoactive compounds in the
mammalian body. Part I: occurrence, origin and metabolism. Med. Biol. 59: 2134.
4. ATLAS, D. & Y. BURNSTEIN. 1984. Isolation and partial purification of clonidine-dis-
placing endogenous brain substance. Eur. J. Biochem. 144: 287293.
5. LI, G., et al. 1994. Agmatine: an endogenous clonidine-displacing substance in the
brain. Science 263: 966969.
6. PRELL, G.D., et al. 2003. Imidazoleacetic acid-ribotide is an endogenous imidazoline
receptor (IR) ligand. Presented at the Fourth International Symposium on Agmatine
and Imidazoline Systems, San Diego, April 2003.
7. ATLAS, D. & Y. BURNSTEIN. 1984. Isolation of an endogenous clonidine-displacing
substance from rat brain. FEBS Lett. 170: 387390.
8. SINGH, G., et al. 1995. Evidence for the presence of a non-catecholamine, clonidine-
displacing substance in crude, methanolic extracts of bovine brain and lung. Naunyn-
Schmiedbergs Arch. Pharmacol. 351: 1726.
9. PARKER, C.A., et al. 1999. Tryptophan: a distinct but biologically inactive component
of clonidine-displacing substance. Br. J. Pharmacol. 127: 72P.
10. MC ISSAC, W.M. 1961. Formation of 1-methyl-6-methoxy-1,2,3,4-tetrahydro-2-carbo-
line under physiological conditions. Biochem. Biophys. Acta 52: 607609.
11. SHOEMAKER, D.W., et al. 1980. Identification of harmane in the rat arcuate nucleus.
Naunyn-Schmiedebergs Arch. Pharmacol. 310: 227230.
12. BIDDER, T.G., et al. 1980. Harmane in human platelets. Life Sci. 25: 157164.
13. ZHENG, W., et al. 2000. Determination of harmane and harmine in human blood using
reversed-phased high performance liquid chromatography and fluorescence detec-
tion. Anal. Biochem. 279: 125129.
14. FEKKES, D. & W.T. BODE. 1993. Occurrence and partition of the -carboline norhar-
mane in rat organs. Life Sci. 52: 20452054.
15. FEKKES, D., et al. 1992. Norharmane, a normal body constituent. Lancet 339: 506.
16. HONECKER, H. & H. ROMMELSPACHER. 1978. Tetrahydroharmane (tetrahydro--carboline),
a physiologically occurring compound of indole metabolism. Naunyn Schmiedebergs
Arch. Pharmacol. 305: 135141.
ROBINSON et al.: ENDOGENOUS b-CARBOLINES 165

17. BARKER, S.A., et al. 1979. Identification and quantification of 1,2,3,4-tetrahydro--carbo-

line, 2-methyl-1,2,3,4-tetrahydro--carboline, and 6-methoxy-1,2,3,4-tetrahydro--
carboline as in vivo constituents of rat brain and adrenal gland. Biochem. Pharmacol.
30: 917.
18. SHOEMAKER, D.W., J.T. CUMMINS & T.G. BIDDER. 1978. -carbolines in rat arcuate
nucleus. Neurosci. 3: 233239.
19. KARI, I., et al. 1983. Mass spectrometric identification of 6-methoxy-1,2,3,4-tetra-
hydro--carboline in pineal gland. In Recent Developments in Biochemistry, Medicine
and Environmental Research. Vol 8: 1924. Elsevier. Amsterdam, The Netherlands.
20. BOSIN, T.R., et al. 1983. Presence of formaldehyde in biological media and organic
solvents: artifactual formation of tetra-hydro--carbolines. Analyt. Biochem. 128:
287293.
21. BROSSI, A. 1993. Mammalian alkaloids II. In The Alkaloids. G.A. Cordell, Ed.: 119
183. Academic Press. New York, NY.
22. ROMMELSPACHER, H., et al. 1976. On the mode of formation of tetrahydro--carbo-
lines. Life Sci. 18: 8188.
23. SUSILO, R. & H. ROMMELSPACHER. 1987. Formation of a -carboline (1,2,3,4-tetra-
hydro-1-methyl--carboline-1-carboxylic acid) following intracerebro-ventricular
injection of tryptamine and pyruvic acid. Naunyn-Schmiedebergs Arch. Pharmacol.
335: 7076.
24. SUSILO, R. & H. ROMMELSPACHER. 1988. Formation of 1-methyl--carbolines in rats
from their possible carboxylica acid precursor. Naunyn-Schmiedebergs Arch. Phar-
macol. 335: 566571.
25. ROMMELSPACHER, H., et al. 1980. 1-Methyl--carboline (harmane) a potent inhibitor of
bezodiadepine receptor binding. Naunyn Schmiedebergs Arch. Pharmacol. 314: 97
100.
26. MORIN, A.M., I.A. TANKAA & C.G. WASTERLAIN. 1981. Norharmane inhibition of [3H]-
diazepam binding in mouse brain. Life Sci. 28: 22572263.
27. MAY, T., et al. 1994. Comparison of the in vitro binding characteristics of the -carbo-
lines harmane and norharmane in rat brain and liver and bovine adrenal medulla.
Naunyn Schmiedebergs Arch. Pharmacol. 349: 308317.
28. KELLER, K.J., et al. 1976. Tryptoline inhibition of serotonin uptake in rat forebrain
homogenates. J. Pharmacol. Exp. Ther. 198: 619625.
29. FAIVRE, V., et al. 2000. Ligand interaction with the purified serotonin transporter in
solution and at the air/water interface. FEBS Lett. 471: 5660.
30. LICHTENBERG-KRAAG, B., et al. 1997. The natural -carbolines facilitate inositol phos-
phate accumulation by activating small G-proteins in human neuroblastoma cells
(SHSY5Y). Neuropharmacology 36: 17711778.
31. KLINKER, J.F., et al. 1997. Activation by -carbolines of G-proteins in HL-60 mem-
branes and the bovine retinal G-protein transducin in a receptor independent manner.
Biochem. Pharmacol. 53: 16211626.
32. STOWOWY, P., R. BONNET & H. ROMMELSPACHER. 1999. The high-affinity binding of
[3H]norharmane ([3H]-carboline) to the ethanol-inducible cytochrome P450 2E1 in
rat liver. Biochem. Pharmacol. 57: 51120.
33. TIARA, Z., et al. 1997. Intercalationof six -carboline derivatives into DNA. Jap. J.
Toxicol. Environ. Health 43: 8391.
34. PAWLIK, M. & H. ROMMELSPACHER. 1988. Demonstration of a distinct class of high-
affinity binding sites for [3H]norharmane ([3H]-carboline) binding sites in the rat
brain. Eur. J. Pharmacol. 147: 163171.
35. GLENNON, R.A., et al. 2000. Binding of -carbolines and related serotonin (5HT2 and
5-HT 1A) dopamine (D2) benzodiazepine receptors. Drug Alcohol Depend. 60: 121
132.
36. MUSGRAVE, I.F. & E. BADOER. 2000. Harmane produces hypotension following micro-
injection into the RVLM: possible role of I1-imidazoline receptors. Br. J. Pharmacol.
129: 10571059.
37. MACKINNES, N. & S.L. HANDLEY. 2002. Characterization of the discriminable stimulus
produced by 2-BFI: effects of imidazoline I2-site ligands, MAOIs, beta-carbolines,
agmatine and ibogaine. Br. J. Pharmacol. 135: 12271234.
166 ANNALS NEW YORK ACADEMY OF SCIENCES

38. ADELL, A., et al. 1996. Action of harmane (1-methyl--carboline) on the brain: body
temperature and in vivo efflux of 5-HT from hippocampus of the rat. Neuropharma-
cology 8: 11011107.
39. HUDSON, A.L., et al. 1999b. Novel selective compounds for the investigation of imida-
zoline receptors. Annals. N. Y. Acad. Sci. 881: 8191.
40. CAPPENDIKJ, S.L.T., D. FEKKES & M.R. DZOLJIC. 1994. The inhibitory effect of norhar-
mane on morphine withdrawal syndrome in rats: comparison with ibogaine. Behav.
Brain Res. 65: 117119.
41. LIONE, L.A., D.J. NUTT & A.L. HUDSON. 1998. Characterisation and localisation of
[3H]2-(2-benzofuranyl)-2-imidazoline binding in rat brain: a selective ligand for imi-
dazoline I2 receptors. Eur. J. Pharmacol. 353: 123135.
42. WIBLE, J.H., et al. 1996. Cardiovascular effects of -carbolines in conscious rats.
Hypertens. Res. 19: 161170.
43. JACKSON, H.C. & D.J. NUTT. 1996. Imidazoline receptors and ingestion. In Drug
Receptor Subtypes and Ingestive Behaviour. Cooper, S.J. & P.G. Clifton, Eds.: 267
83. Academic Press. London.
44. MORGAN, N.G., et al. 2003. Comparative effects of efaroxan and b-carbolines on the
secretory activity of rodent and human b cells. Ann. N. Y. Acad. Sci. This volume.
45. EDWARDS, M.M. 2003. Imidazoline ligands and feeding behaviour: the role of imida-
zoline binding sites. Ph.D. thesis, University of Bristol, Bristol, UK.
46. ROMMELSPACHER, H., et al. 1977. Pharmacological properties of tetrahydronorharmane
(tryptoline). Naunyn-Schmiederbergs Arch. Pharmacol. 298: 8391.
47. COLLINS, M.A. 2002. Alkaloids, alcohol and Parkinsons disease. Parkinsonism Relat.
Disord. 8: 417422.
48. DE MEESTER, C. 1995. Genotoxic potential of -carbolines: a review. Mutation Res.
Rev. Gen. Toxicol. 7. 339: 139153.
49. GEARHART, D.A., et al. 2000. Increased -carboline 9N-methyltransferase activity in
the frontal cortex in Parkinsons disease. Neurobiol. Dis. 7: 201211.
50. GEARHART, D.A., P.F. TOOLE & J.W. BEEACH. 2002. Identification of brain proteins that
interact with 2-methylnorharman. An analog of the Parkinsonian-inducing toxin,
MPP+. Neurosci. Res. 44: 255265.