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J Biol Chem, Vol. 273, Issue 36, 23321-23326, September 4, 1998
From the Departments of Molecular Biology and Pharmacology,
Synaptic Pharmaceutical Corporation, Paramus, New Jersey 07652
The neuropeptide galanin has been implicated in
the regulation of processes such as nociception, cognition, feeding
behavior, and hormone secretion. Multiple galanin receptors are
predicted to mediate its effects, but only two functionally coupled
receptors have been reported. We now report the cloning of a third
galanin receptor distinct from GALR1 and GALR2. The receptor, termed
GALR3, was isolated from a rat hypothalamus cDNA library by both
expression and homology cloning approaches. The rat GALR3 receptor
cDNA can encode a protein of 370 amino acids with 35% and 52%
identity to GALR1 and GALR2, respectively. Localization of mRNA by
solution hybridization/RNase protection demonstrates that the GALR3
transcript is widely distributed, but expressed at low abundance, with
the highest levels in the hypothalamus and pituitary. We also isolated the gene encoding the human homologue of GALR3. The human GALR3 receptor is 90% identical to rat GALR3 and contains 368 amino acids.
Binding of porcine 125I-galanin to stably expressed
rat and human GALR3 receptors is saturable (rat KD = 0.98 nM and human KD = 2.23 nM) and displaceable by galanin peptides and analogues in
the following rank order: rat galanin, porcine galanin The neuropeptide galanin modulates a variety of physiological
processes including cognition/memory (1), sensory/pain processing (2,
3), hormone secretion (4, 5), and feeding behavior (6, 7) (for review,
see Ref. 8). The endogenous and exogenous effects of galanin are
mediated by multiple receptor subtypes. To date, two cloned galanin
receptors, GALR1 and GALR2, have been shown to functionally couple to
their intracellular effectors through distinct signaling pathways:
GALR1 inhibits adenylyl cyclase via a pertussis toxin-sensitive
G-protein of the Gi/Go family (9-11), whereas
GALR2 stimulates inositol phospholipid turnover and intracellular
calcium mobilization through a pertussis toxin-insensitive Gq/G11-type mechanism (11, 12). The
pharmacological profiles of GALR1 and GALR2 are similar to one another
in that both show a preference for the N-terminal portion of the
galanin peptide, and chimeric "antagonist" peptides such as M35 and
M40 act as full agonists at both cloned receptors (11, 13). These
properties of GALR1 and GALR2 do not fully explain aspects of galanin
pharmacology such as the reported agonist activity of galanin-(3-29)
in the pituitary (14), guinea pig stomach (15, 16), and hypothalamus (17) or the antagonist properties of the chimeric galanin analogues M15, M35, and M40 in physiological models of galanin-mediated luteinizing hormone-releasing hormone release (18), facilitation of the
spinal flexor reflex (19), and feeding (7, 20), respectively (for
review, see Ref. 21). Together, these data suggested that additional
galanin receptor subtypes beyond GALR1 and GALR2 existed. Recently, the
cloning of a third galanin receptor cDNA was described by Wang
et al. (22), but the pharmacological characterization of
this receptor was limited by an extremely low apparent
Bmax in porcine 125I-galanin binding
assays, and no functional coupling was reported.
We now report the cloning and functional characterization of a third
galanin receptor subtype, termed GALR3, from the rat hypothalamus based
on a combination of expression and homology cloning methods. We have
also isolated and characterized the functional human homologue of
GALR3. Both human and rat GALR3 receptors exhibit pharmacological
profiles distinct from GALR2 and GALR1. Moreover, we have shown that
galanin acting at GALR3 receptors can activate GIRK currents in
Xenopus oocytes, demonstrating that GALR3 can couple to a
G-protein of the Gi/Go class. The cloning and
characterization of rat and human GALR3 receptors will facilitate the
association of functional roles with specific galanin receptor subtypes
and will help to circumscribe the properties of any remaining uncloned galanin receptors.
Isolation of a Novel Rat Galanin Receptor: Convergence of
Homology and Expression Cloning--
A rat hypothalamus cDNA
library in
Cloned Human and Rat Galanin GALR3 Receptors
PHARMACOLOGY AND ACTIVATION OF G-PROTEIN INWARDLY RECTIFYING
K+ CHANNELS*
,
ABSTRACT
Top
Abstract
Introduction
Procedures
Results & Discussion
References
M32,
M35 porcine galanin-(
7 to +29), galantide, human
galanin > M40, galanin-(1-16) > [D-Trp2]galanin-(1-29),
galanin-(3-29). This profile resembles that of the rat
GALR1 and GALR2 receptors with the notable exception that human
galanin, galanin-(1-16), and M40 show lower affinity at GALR3. In
Xenopus oocytes, activation of rat and human GALR3
receptors co-expressed with potassium channel subunits GIRK1 and GIRK4
resulted in inward K+ currents characteristic of
Gi/Go-coupled receptors. These data confirm the
functional efficacy of GALR3 receptors and further suggest that GALR3
signaling pathways resemble those of GALR1 in that both can activate
potassium channels linked to the regulation of neurotransmitter
release.
INTRODUCTION
Top
Abstract
Introduction
Procedures
Results & Discussion
References
EXPERIMENTAL PROCEDURES
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Abstract
Introduction
Procedures
Results & Discussion
References
ZAP II (Stratagene, La Jolla, CA) was screened using
overlapping oligonucleotide probes representing the transmembrane
domains of the rat GALR2 receptor cDNA (11). The oligomers were
labeled, and hybridizations were performed at reduced stringency as
described (23). The cDNA insert from one hybridizing clone, rHy35a,
was sequenced on both strands by cycle sequencing with AmpliTaq DNA
polymerase FS (Perkin-Elmer), and products were run on an automated
fluorescent sequencer (Applied Biosystems, Inc.). Sequence analysis
(GCG Version 9.1, Genetics Computer Group, Inc., Madison, WI) indicated
that rHy35a represented a fragment of a novel galanin-like
receptor.
Cloning and Sequencing a Novel Human Galanin Receptor
Fragment--
A human placenta genomic library in DASH II
(
1.5 × 106 total recombinants; Stratagene) was
screened at reduced stringency using GALR2 transmembrane domain probes
as described above. One positive phage clone, plc21a, contained a
2.7-kilobase pair KpnI/EcoRI fragment that
hybridized with the rat GALR2 transmembrane domain II oligonucleotide
probe. Subcloning and sequence analysis revealed that the fragment
encoded the first three transmembrane domains of the human GALR3
receptor as well as a large intron. To obtain additional exons encoding
the receptor, the original phage clone plc21a (~18-kilobase pair
insert) was hybridized with probes directed to the third extracellular
loop or transmembrane IV regions of the rat GALR3 cDNA. The
full-length human GALR3 construct was prepared by ligating a polymerase
chain reaction-derived product of the 5'-exon, representing the
starting Met through the III/IV loop, with a genomic fragment
containing the 3'-exon.
Cell Culture and Expression--
The calcium phosphate technique
(25) was used to co-transfect receptor cDNA plus a G418-resistant
plasmid into mouse fibroblast LMTK
cells for human GALR3
studies or into human embryonic kidney 293 cells for rat GALR3 studies.
Stable clones were selected with G418 and screened as membrane
preparations for specific porcine 125I-galanin binding. By
the same protocol, rat GALR1 receptors were stably expressed in Chinese
hamster ovary cells, and rat GALR2 receptors in LMTK
cells. Supplements (10% bovine calf serum, 4 mM
L-glutamine, and 100 units/ml penicillin and 100 µg/ml
streptomycin) were added to Dulbecco's modified Eagle's medium for
the culture of mouse fibroblast LMTK and human embryonic
kidney 293 cells or to Ham's F-12 medium for Chinese hamster ovary
cells. Cells were grown on 150-mm plates at 37 °C and 5%
CO2. Stock plates were split with trypsin every 3-4 days
at a ratio of 1:6 for 293 cells, 1:8 for Chinese hamster ovary cells,
or 1:10 for LMTK cells.
Radioligand Binding Assay-- Membranes were prepared from transfected cells, and protein was measured as described previously (11). Membranes, peptides, and porcine 125I-galanin (2200 Ci/mmol; NEN Life Science Products) were diluted in galanin binding buffer supplemented with 0.1% bovine serum albumin and 0.1% bacitracin and then distributed into 96-well Millipore GF/C filter plates (pretreated for 30 min with 1% polyethyleneimine in 50 mM Tris, pH 7.4) so that the 125I-galanin concentration was ~375,000 dpm/sample for GALR3 and ~187,500 dpm/sample for GALR1 and GALR2. Following incubation at room temperature for 120 min, membranes were collected by filtration, and 125I was measured by liquid scintillation. Nonspecific binding was defined by 1 µM porcine galanin. Saturation and displacement binding data were analyzed by nonlinear regression and are reported as mean ± S.E.
Oocyte Injection and Recording-- The sequences comprising the coding regions of rat and human GALR3 were subcloned into pBS KS+AMV-pA50 (26) modified to contain a SrfI site. Genes encoding GIRK1 and GIRK4 were obtained by polymerase chain reaction from human heart cDNA using published sequences (27-29) to derive appropriate forward and reverse amplimers. All mRNAs were transcribed using T7 polymerase (Message Machine, Ambion Inc.). Oocytes received 2 ng each of GIRK1 and GIRK4 mRNAs in combination with 5-25 ng of receptor mRNA; after injection, oocytes were incubated at 17 °C for 3-8 days. Dual-electrode voltage clamping was performed using microelectrodes with resistances of 1-3 megaohms. During the application of agonists, oocytes were superfused in high K+ medium containing 48 mM KCl, 49 mM NaCl, 2 mM CaCl2, 2 mM MgCl2, and 5 mM HEPES, pH 7.5.
Solution Hybridization/Nuclease Protection Assay--
Solution
hybridization/ribonuclease protection assays were used to detect rat
GALR3 receptor transcripts in mRNA isolated from rat tissues. A
445-base pair fragment of the rat GALR3 cDNA (nucleotides
1061-1506) flanked by RNA polymerase promoter sequences was used to
synthesize a radiolabeled GALR3 cRNA probe using standard methods and
reagents (Promega). Solution hybridization assays were performed as
described previously (30). Autoradiographs of assays were generated by
apposing dried gels to PhosphorImager screens (Molecular Dynamics,
Inc.) or Eastman Kodak BioMax film at 70 °C.
Materials-- Cell culture media and supplements were from Specialty Media (Lavallette, NJ). Cell culture plates (150-mm and 96-well microtiter) were from Corning (Corning Inc., NY). Ex-Cell 400TM medium with L-glutamine was purchased from JRH Biosciences. Polypropylene 96-well microtiter plates were from Costar Corp. (Cambridge, MA). All radioligands were from NEN Life Science Products. Galanin and related peptide analogues were from either Bachem California (Torrance, CA) or Peninsula Laboratories, Inc. (Belmont, CA) or were synthesized by custom order from Chiron Mimotopes Peptide Systems (Clayton, Victoria, Australia). Bio-Rad reagent was from Bio-Rad. Bovine serum albumin (ultra fat-free, A-7511) was from Sigma. All other materials were reagent-grade.
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RESULTS AND DISCUSSION |
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Results & Discussion
References
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Cloning of Rat and Human GALR3 Receptors-- Recent evidence indicated that GALR1 and GALR2 receptor mRNAs were present in the rat hypothalamus (10, 11, 31), but not all aspects of the cloned GALR1 and GALR2 pharmacological profiles matched those observed for galanin-mediated feeding (7, 20) as well as other physiological effects of galanin analogues (14, 21, 32). Based on the potential presence of multiple galanin receptors in the hypothalamus, we attempted to isolate additional galanin receptors using both homology and expression cloning strategies. The convergence of these strategies resulted in the isolation of a partial galanin receptor-like cDNA fragment (rHy35a) by reduced stringency homology cloning and the discovery of the same fragment in a galanin-binding library pool (K163, ~3200 primary clones) identified by expression cloning. Isolation of the full-length cDNA from pool K163 of the rat hypothalamus expression library and subsequent transfection/radioligand binding confirmed that the cDNA encoded a novel rat galanin receptor, termed GALR3. The human homologue of GALR3 was subsequently isolated from a human placenta genomic library and subcloned for sequencing and expression studies.
The rat GALR3 cDNA can encode a protein of 370 amino acids with 35% amino acid identity to rat GALR1 (10) and 52% identity to rat GALR2 (11), the receptor to which it is most closely related. The sequence similarity to GALR2 is higher within transmembrane domains II-IV, where amino acid identities range from 70 to >90%. The human GALR3 receptor comprises 368 amino acids with 90% identity to rat GALR3 (Fig. 1); compared with human galanin receptor subtypes, GALR3 is 37 and 57% identical to human GALR1 (9) and human GALR2 (33), respectively. In accord with its high level of sequence identity to GALR2, the human GALR3 gene contains an intron within the coding region at the same location as reported for rat GALR2 (11, 34). Notably, the amino acid sequences for the rat and human GALR3 receptors shown here (Fig. 1) differ from previous preliminary reports: the sequence of human GALR3 differs by 6 amino acids from that of an unannotated receptor-like sequence in GenBankTM (EM_HTG:HS81I2), and the rat GALR3 receptor differs by 4 amino acids from another galanin receptor also described as GALR3 by Wang et al. (22). Both rat and human GALR3 receptor sequences contain a single consensus site for N-linked glycosylation in the N terminus (see Fig. 1) and several predicted intracellular sites for phosphorylation by protein kinases; one putative phosphorylation site common to rat and human GALR3 in the third intracellular loop is absent in GALR1 and GALR2.
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Pharmacological Characterization of Rat and Human GALR3 Receptors-- Radioligand binding assays were conducted to characterize the cloned GALR3 receptor homologues. Porcine 125I-galanin bound in specific and saturable fashion to membranes from cells stably transfected with either human or rat GALR3. Data generated from experiments in which porcine 125I-galanin concentrations ranged from 0.5 pM to 3.0 nM were best fit to a one-site model. Apparent pKd and Bmax values are listed in Table I. For comparison, rat GALR1 and GALR2 receptors were studied under similar conditions. Data in Table I show that both the observed binding affinity of porcine 125I-galanin and the measurable Bmax were lowest for the GALR3 receptors.
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7 to +29) > human
galanin, porcine galanin-(1-16) > porcine galanin-(3-29). The
chimeric peptides bound to rat GALR3 with a distinctive rank order:
M32, C7, M35, galantide > M40. Comparison with the other cloned
rat galanin receptor subtypes indicates that apart from quantitative
differences, rat GALR3 shares certain features in common with rat GALR1
despite the high degree of sequence divergence. For example, both
receptor subtypes display the lowest affinity for M40 out of all the
chimeric peptides studied, and both receptors are sensitive to galanin
C-terminal truncation, with lower affinity for porcine galanin-(1-16)
versus porcine galanin. Also, GALR3 and GALR1 receptors are
sensitive to N-terminal modification in having decreased affinity for
[D-Trp2]galanin versus galanin and
no detectable binding affinity for porcine galanin-(3-29), the latter
being a common feature of all three cloned galanin receptor subtypes.
In accord with its high level of amino acid identity to GALR2, GALR3
maintains moderate affinity for galanin-(2-29) (22). In sharing a
general preference for the conserved N-terminal portion of the galanin
peptide, all three cloned receptors are thus distinct from the galanin
receptor subtypes activated by C-terminal fragments of galanin (14, 16, 17, 38) as well as galanin-(1-15)-preferring subtypes in the dorsal
hippocampus and brain stem (39, 40).
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3).
Functional Coupling of the GALR3 Receptor--
Clues about GALR3
function may be obtained from native systems. There is support for
inhibitory effects of galanin on neurotransmitter/hormone release in
many tissues expressing GALR3 mRNA, including the hypothalamus (42), pituitary (43), spinal cord (44), and pancreas (45), commonly
through inhibition of voltage-sensitive Ca2+ channels.
G-protein antisense studies of pituitary-derived GH3 cells and
pancreas-derived RINm5F cells, involving Ca2+ channel
inhibition, indicated that galanin signals preferentially through
G
o1
2
2 in those cells (46).
Galanin is also proposed to inhibit neurotransmitter release through
activation of ATP-sensitive K+ channels and inwardly
rectifying K+ channels (GIRK channels) (47).
24 mV) close to the predicted equilibrium potential for
K+ (23 mV in high K+ medium), 4) sensitivity
to blockade by 300 µM Ba2+, and 5) lack of
galanin sensitivity in oocytes injected with only GALR3 mRNA (data
not shown). Injection of pertussis toxin (2 ng/oocyte) caused a 98%
reduction (n = 10) of galanin currents in oocytes
expressing GALR3, supporting the conclusion that, in oocytes, GALR3
receptors couple to a pertussis toxin-sensitive Gi/Go pathway. This functionality is
similar to GALR1 receptors, which also activate GIRK channels (Fig.
4C) in a pertussis
toxin-sensitive manner (data not shown), but is different from GALR2
receptors, which, in oocytes, activate inward Cl currents
characteristic of Gq-coupled receptors (11). Thus, the
cloned GALR3 receptor appears to use a G-protein of the
Gi/Go class to modulate a channel (GIRK)
associated with hyperpolarization and inhibition of neurotransmitter
release.
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7 to +29), galanin-(1-16), and M40 evoked
agonist activity at a fixed dose of 1 µM;
[D-Trp2]galanin and galanin-(3-29) were
inactive. These last two peptides were also tested for their ability to
block galanin-evoked currents, but no antagonism was detected.
EC50 values obtained for selected peptides from cumulative
concentration-response measurements (Fig. 4B) were 56 nM for M32, 206 nM for porcine galanin, 343 nM for C7, 1907 nM for human galanin, 1678 nM for galanin(7 to +29), 3270 nM for
galanin-(1-16), and 3999 nM for M40. The rank order of
potency was similar to that observed for displacement of
125I-galanin in binding assays using LMTK
cells stably expressing human GALR3 (Table II). The apparent overall
low potency of galanin and related peptides in oocytes did not seem to
be related to a low efficiency of receptor coupling inherent to oocytes
since galanin exhibited an EC50 of 2 nM in oocytes expressing GALR1 (Fig. 4C). Instead, the low potency
may reflect the somewhat lower affinity of galanin peptides for GALR3 (Tables I and II). Overall, the electrophysiological data reveal a
functional similarity between GALR1 and GALR3 in oocytes despite a low
level of primary sequence identity; exactly which G-proteins are
involved in the oocyte or would be involved in mammalian cells remain
to be determined for each receptor subtype. We are now in the process
of extrapolating these findings from the oocyte to the study of GALR3
receptors transfected into mammalian cells.
The localization, functional coupling, and pharmacology of GALR3
suggest a number of physiological roles for this receptor in the
regulation of feeding, inhibition of neurotransmitter release (i.e. acetylcholine, serotonin, and norepinephrine),
regulation of pituitary endocrine release, inhibition of
glucose-stimulated insulin release, and regulation of spinal cord
excitability (47). The involvement of GALR3 mRNA in spinal cord
function is particularly intriguing. GALR3 shows high binding affinity
(relative to galanin) for the alternately processed galanin-(7 to
+29). This peptide and galanin-(9 to + 29) are found in the adrenal
gland (48) and modulate spinal excitability, albeit with weaker potency
than full-length galanin (49). Furthermore, a
galanin-dependent inward current appears in dorsal root
ganglia only after axotomy, when galanin mRNA is up-regulated, but
GALR1 and GALR2 mRNAs are decreased (50, 51). It would be
interesting to determine whether the novel current is mediated by GALR3
or supports the existence of additional galanin receptor subtypes.
The ability of the chimeric peptides to bind GALR3 (M32, C7, M35,
galantide > M40) and to activate GALR3 in oocytes prompts a
re-evaluation of the actions of these peptides. The chimeras were
classified as antagonists in tissues such as hypothalamus, hippocampus,
pituitary, spinal cord, and pancreas (21, 52), but are agonists for the
cloned GALR1, GALR2, and GALR3 receptors (Refs. 11 and 13 and this
paper). The discrepancy may be explained by a
system-dependent spectrum of partial agonist/antagonist
activity, or the chimeric peptides may be degraded in vivo
to fragments having different properties than the parent peptides.
Alternatively, it may also indicate the existence of additional
receptor subtypes. The pharmacology of cloned GALR1, GALR2, and GALR3
receptors cannot explain the binding of galanin-(3-29) to a receptor
in the pituitary (14) or gastric smooth muscle (16), nor can it explain
a galanin-(1-15)-preferring receptor in the hippocampus (39) or locus
ceruleus (40). Continued cloning efforts, combined with the
identification of stable subtype-selective ligands, should help define
the contributions of GALR1, GALR2, and GALR3 to the broad picture of
galanin-dependent physiology.
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ACKNOWLEDGEMENTS |
|---|
For expert technical assistance, we thank Michelle Smith, Nancy Shen, Steve Fried, Sherif Daouti, Ling-Yan Huang, Mary Johnson, Harvey Lichtblau, Hui-Ying Chen, and Meng Dai (Department of Molecular Biology, Synaptic Pharmaceutical Corp.); Zoya Shaposhnik, Marc Dizon, Rong Zhou, Raisa Nagorny, and Steven Valerio (Department of Pharmacology); and Tracy Johnson-Blake, Debbie Tambe, Tracy Taylor, Rosa Panico, Enrique Hurtado, Eddie Nerio, and Staci Baratoglou (Department of Cell Biology). We also thank Dr. Angelica Bonin for cell and molecular biology support and George Moralishvili for excellent artwork and illustrations.
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FOOTNOTES |
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* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF073798 and AF073799.
To whom correspondence should be addressed: Synaptic
Pharmaceutical Corp., 215 College Rd., Paramus, NJ 07652. Tel.:
201-261-1331 (ext. 685); Fax: 201-261-0623; E-mail:
ksmith{at}synapticcorp.com.
§ Current address: Wyeth-Ayerst Research, CNS Dept., 865 Ridge Rd., Monmouth Junction, NJ 08852.
The abbreviation used is: GMAP, galanin message-associated peptide.
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REFERENCES |
|---|
|
Top
Abstract
Introduction
Procedures
Results & Discussion
References
|
|---|
- Crawley, J. N. (1996) Life Sci. 58, 2185-2199[CrossRef][Medline] [Order article via Infotrieve]
-
Wiesenfeld-Hallin, Z.,
Xu, X. J.,
Langel, U.,
Bedecs, K.,
Hokfelt, T.,
and Bartfai, T.
(1992)
Proc. Natl. Acad. Sci. U. S. A.
89,
3334-3337
[Abstract/Free Full Text] - Ji, R. R., Zhang, X., Zhang, Q., Dagerlind, A., Nilsson, S., Wiesenfeld-Hallin, Z., and Hokfelt, T. (1995) Neuroscience 68, 563-576[CrossRef][Medline] [Order article via Infotrieve]
- Chan, Y. Y., Grafstein-Dunn, E., Delemarre-Van de Waal, H. A., Burton, K. A., Clifton, D. K., and Steiner, R. A. (1996) Endocrinology 137, 5303-5310[Abstract]
- Grottoli, S., Arvat, E., Gianotti, L., Ramunni, J., Di Vito, L., Maccagno, B., Ciccarelli, E., Camanni, F., and Ghigo, E. (1996) J. Endocrinol. Invest. 19, 739-744[Medline] [Order article via Infotrieve]
- Crawley, J. N., Austin, M. C., Fiske, S. M., Martin, B., Consolo, S., Berthold, M., Langel, U., Fisone, G., and Bartfai, T. (1990) J. Neurosci. 10, 3695-3700[Abstract]
- Corwin, R. L., Robinson, J. K., and Crawley, J. N. (1993) Eur. J. Neurosci. 5, 1528-1533[CrossRef][Medline] [Order article via Infotrieve]
-
Evans, H. F.,
and Shine, J.
(1991)
Endocrinology
129,
1682-1684
[Abstract/Free Full Text] -
Habert-Ortoli, E.,
Amiranoff, B.,
Loquet, I.,
Laburthe, M.,
and Mayaux, J. F.
(1994)
Proc. Natl. Acad. Sci. U. S. A.
91,
9780-9783
[Abstract/Free Full Text] - Parker, E. M., Izzarelli, D. G., Nowak, H. P., Mahle, C. D., Iben, L. G., Wang, J., and Goldstein, M. E. (1995) Brain Res. Mol. Brain Res. 34, 179-189[Medline] [Order article via Infotrieve]
-
Smith, K. E.,
Forray, C.,
Walker, M. W.,
Jones, K. A.,
Tamm, J. A.,
Bard, J.,
Branchek, T. A.,
Linemeyer, D. L.,
and Gerald, C.
(1997)
J. Biol. Chem.
272,
24612-24616
[Abstract/Free Full Text] - Fathi, Z., Cunningham, A. M., Iben, L. G., Battaglino, P. B., Ward, S. A., Nichol, K. A., Pine, K. A., Wang, J. C., Goldstein, M. E., Iismaa, T. P., and Zimanyi, I. A. (1997) Mol. Brain Res. 51, 49-59[Medline] [Order article via Infotrieve]
- Forray, C., Smith, K. E., Gerald, C., Vaysse, P., Weinshank, R. L., and Branchek, T. A. (1996) Soc. Neurosci. Abstr. 22, 1304
-
Wynick, D.,
Smith, D. M.,
Ghatei, M.,
Akinsanya, K.,
Bhogal, R.,
Purkiss, P.,
Byfield, P.,
Yanaihara, N.,
and Bloom, S. R.
(1993)
Proc. Natl. Acad. Sci. U. S. A.
90,
4231-4235
[Abstract/Free Full Text] -
Gu, Z. F.,
Pradhan, T. K.,
Coy, D. H.,
and Jensen, R. T.
(1994)
Am. J. Physiol.
266,
G839-G845
[Abstract/Free Full Text] -
Gu, Z. F.,
Pradhan, T. K.,
Coy, D. H.,
and Jensen, R. T.
(1995)
J. Pharmacol. Exp. Ther.
272,
371-378
[Abstract/Free Full Text] -
Kinney, G. A.,
Emmerson, P. J.,
and Miller, R. J.
(1998)
J. Neurosci.
18,
3489-3500
[Abstract/Free Full Text] -
Sahu, A.,
Xu, B.,
and Kalra, S. P.
(1994)
Endocrinology
134,
529-536
[Abstract/Free Full Text] - Xu, X. J., Wiesenfeld-Hallin, Z., Langel, U., Bedecs, K., and Bartfai, T. (1995) Br. J. Pharmacol. 116, 2076-2080[Medline] [Order article via Infotrieve]
- Crawley, J. N., Robinson, J. K., Langel, U., and Bartfai, T. (1993) Brain Res. 600, 268-272[CrossRef][Medline] [Order article via Infotrieve]
- Bartfai, T., Fisone, G., and Langel, U. (1992) Trends Pharmacol. Sci. 13, 312-317[CrossRef][Medline] [Order article via Infotrieve]
-
Wang, S. K.,
He, C. G.,
Hashemi, T.,
and Bayne, M.
(1997)
J. Biol. Chem.
272,
31949-31952
[Abstract/Free Full Text] -
Bard, J. A.,
Walker, M. W.,
Branchek, T. A.,
and Weinshank, R. L.
(1995)
J. Biol. Chem.
270,
26762-26765
[Abstract/Free Full Text] - McCormick, M. (1987) Methods Enzymol. 151, 445-449[Medline] [Order article via Infotrieve]
- Cullen, B. R. (1987) Methods Enzymol. 152, 684-704[Medline] [Order article via Infotrieve]
-
Nowak, M. W.,
Kearney, P. C.,
Sampson, J. R.,
Saks, M. E.,
Labarca, C.,
Silverman, S. K.,
Zhong, W.,
Thorson, J.,
Abelson, J. N.,
and Davidson, N.
(1995)
Science
268,
439-442
[Abstract/Free Full Text] - Kubo, Y., Reuveny, E., Slesinger, P. A., Jan, Y. N., and Jan, L. Y. (1993) Nature 364, 802-806[CrossRef][Medline] [Order article via Infotrieve]
-
Dascal, N.,
Schreibmayer, W.,
Lim, N. F.,
Wang, W.,
Chavkin, C.,
DiMagno, L.,
Labarca, C.,
Kieffer, B. L.,
Gaveriaux-Ruff, C.,
and Trollinger, D.
(1993)
Proc. Natl. Acad. Sci. U. S. A.
90,
10235-10239
[Abstract/Free Full Text] - Krapivinsky, G., Gordon, E. A., Wickman, K., Velimirovic, B., Krapivinsky, L., and Clapham, D. E. (1995) Nature 374, 135-141[CrossRef][Medline] [Order article via Infotrieve]
- Artymyshyn, R. P., Ivins, K. J., Monks, B. R., Luedtke, R. R., and Molinoff, P. B. (1992) Neurochem. Int. 20, 189S-195S
- Gustafson, E. L., Smith, K. E., Durkin, M. M., Gerald, C., and Branchek, T. A. (1996) Neuroreport 7, 953-957[Medline] [Order article via Infotrieve]
-
Bartfai, T.,
Langel, U.,
Bedecs, K.,
Andell, S.,
Land, T.,
Gregersen, S.,
Ahren, B.,
Girotti, P.,
Consolo, S.,
and Corwin, R.
(1993)
Proc. Natl. Acad. Sci. U. S. A.
90,
11287-11291
[Abstract/Free Full Text] - Borowsky, B., Smith, K. E., Bard, J. A., Walker, M. W., Huang, L.-Y., Branchek, T. A., and Gerald, C. (1997) Soc. Neurosci. Abstr. 22, 393
- Howard, A. D., Tan, C., Shiao, L. L., Palyha, O. C., McKee, K. K., Weinberg, D. H., Feighner, S. D., Cascieri, M. A., Smith, R. G., Van Der Ploeg, L. H. T., and Sullivan, K. A. (1997) FEBS Lett. 405, 285-290[CrossRef][Medline] [Order article via Infotrieve]
- Chan-Palay, V. (1990) Adv. Neurol. 51, 253-255[Medline] [Order article via Infotrieve]
- Schreiber, R. C., Hyatt-Sachs, H., Bennett, T. A., and Zigmond, R. E. (1994) Neuroscience 60, 17-27[CrossRef][Medline] [Order article via Infotrieve]
- Xu, Z. Q., Shi, T. J., Landry, M., and Hoekfelt, T. (1996) Neuroreport 8, 237-242[Medline] [Order article via Infotrieve]
- Chakder, S., and Rattan, S. (1991) Gastroenterology 100, 711-718[Medline] [Order article via Infotrieve]
- Hedlund, P. B., Finnman, U. B., Yanaihara, N., and Fuxe, K. (1994) Brain Res. 634, 163-167[CrossRef][Medline] [Order article via Infotrieve]
- Diaz, Z., Narvaez, J. A., Hedlund, P. B., Aguirre, J. A., Gonzalez-Baron, S., and Fuxe, K. (1996) Brain Res. 741, 32-37[CrossRef][Medline] [Order article via Infotrieve]
- Wang, Z. L., Kulkarni, R. N., Wang, R. M., Smith, D. M., Ghatei, M. A., Byfield, P. G. H., Bennet, W. M., and Bloom, S. R. (1997) J. Clin. Invest. 100, 189-196[Medline] [Order article via Infotrieve]
-
Tsuda, K.,
Yokoo, H.,
and Goldstein, M.
(1989)
Hypertension (Dallas)
14,
81-86
[Abstract/Free Full Text] - Cimini, V. (1996) Exp. Cell Res. 228, 212-215[CrossRef][Medline] [Order article via Infotrieve]
- Reimann, W., and Schneider, F. (1993) Life Sci. 52, PL251-PL254[CrossRef][Medline] [Order article via Infotrieve]
- Renstrom, E., Ding, W. G., Bokvist, K., and Rorsman, P. (1996) Neuron 17, 513-522[CrossRef][Medline] [Order article via Infotrieve]
- Kalkbrenner, F., Degtiar, V. E., Schenker, M., Brendel, S., Zobel, A., Heschler, J., Wittig, B., and Schultz, G. (1995) EMBO J. 14, 4728-4737[Medline] [Order article via Infotrieve]
- Kask, K., Berthold, M., and Bartfai, T. (1997) Life Sci. 60, 1523-1533[CrossRef][Medline] [Order article via Infotrieve]
-
Bersani, M.,
Thim, L.,
Rasmussen, T. N.,
and Holst, J. J.
(1991)
Endocrinology
129,
2693-2698
[Abstract/Free Full Text] - Bedecs, K., Langel, U., Xu, X. J., Wiesenfeld-Hallin, Z., and Bartfai, T. (1994) Eur. J. Pharmacol. 259, 151-156[CrossRef][Medline] [Order article via Infotrieve]
-
Xu, Z. Q.,
Zhang, X.,
Grillner, S.,
and Hokfelt, T.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
13262-13266
[Abstract/Free Full Text] - Sten Shi, T. J., Zhang, X., Holmberg, K., Xu, Z. Q., and Hokfelt, T. (1997) Neurosci. Lett. 237, 57-60[CrossRef][Medline] [Order article via Infotrieve]
- Kask, K., Langel, U., and Bartfai, T. (1995) Cell. Mol. Neurobiol. 15, 653-673[CrossRef][Medline] [Order article via Infotrieve]
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