Gene structure and expression of the mouse 5-HT2 receptor.
Serotonin (5-hydroxytryptamine, 5-HT) is an important neurotransmitter which mediates numerous physiological functions. Using the SacI-EcoRI restriction fragment of the rat brain 5-HT2 receptor cDNA as a probe, we have screened a mouse brain cDNA library, created by random priming and constructed in SWAJ vectors, and have isolated a cDNA encoding a 1.4 kb open reading frame which codes for a functional mouse 5-HT2 receptor identified from pharmacological binding profiles and coupling of phosphoinositide formation in a stably transfected fibroblast cell line. The deduced amino acid sequence is 97.4% identical to the rat 5-HT2 receptor. Using the same 5-HT2 receptor cDNA probe, ten positive genomic clones were isolated from two mouse genomic libraries constructed in the pWE15 cosmid vector and the EMBL-3 phage vector. Extensive mapping and sequencing of these genomic clones indicate the mouse 5-HT2 receptor coding region spans over 20 kb and is composed of three exons split by two introns. Northern blot analysis shows one band of 5-6 kb in the mouse brain, but not in the heart, lung, liver, or kidney total RNA. Southern analysis of mouse liver genomic DNA shows a simple pattern of digestion by several restriction enzymes, which suggests that one copy of the 5-HT2 receptor gene may exist in the mouse genome.
Yang
W
W
Department of Molecular Pharmacology and Toxicology, School of Pharmacy, University of Southern California, Los Angeles 90033.
Chen
K
K
Lan
N C
NC
Gallaher
T K
TK
Shih
J C
JC
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000502:physiology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D015723:Gene Library; D005796:Genes; D007650:Ketanserin / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010716:Phosphatidylinositols / Q000378:metabolism; D011985:Receptors, Serotonin / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection
D010716:Phosphatidylinositols; D011985:Receptors, Serotonin; D004247:DNA; D007650:Ketanserin