Structure and expression of the rat neuropeptide Y gene.

Abstract

Neuropeptide Y is a 36-amino acid peptide that is abundant throughout the mammalian nervous system. It belongs to the same family of carboxyl-terminally amidated peptides as pancreatic polypeptide and peptide YY. We describe here the gene encoding the rat neuropeptide Y precursor. The gene spans 7.2 kilobase pairs and contains four exons. The exon organization is identical to the pancreatic polypeptide gene, although the amino acid sequences of the neuropeptide Y and pancreatic polypeptide precursors differ extensively. The predicted amino acid sequence of mature rat neuropeptide Y is identical to the human sequence. Also the sequence of the 30-amino acid carboxyl-terminal peptide of preproneuropeptide Y is highly conserved, which suggests that it is functionally important. Two neuropeptide Y alleles were found to differ at nine positions in 2.5 kilobase pairs at the 5' portion of the gene. No exon difference was found. One nucleotide substitution close to the gene promoter may influence the regulation of expression. Neuropeptide Y mRNA was found in all rat brain subregions tested, which shows that neuropeptide Y is synthesized throughout the brain. Developmentally, mRNA was detected in the rat brain as early as embryonic day 16 and increased rapidly to adult levels. The level of neuropeptide Y mRNA was also studied in several rat peripheral organs. Unexpectedly high levels were observed in heart and spleen. This mRNA may be synthesized in intrinsic ganglia and non-neuronal cells, respectively.Neuropeptide Y is one of the most abundant and widespread peptides in the mammalian nervous system (1, 2). It is a 36-amino acid peptide that shares considerable sequence homology with the endocrine gut peptides pancreatic polypeptide and peptide YY. All three members of this peptide family have a carboxyl-terminal amide group. This property initially led to the discovery of neuropeptide Y and peptide YY (3,4). The widespread distribution of neuropeptide Y in the brain and the findings that it coexists with catecholamine transmitters in many neurones (5) and in adrenal medulla (6) suggest that this peptide serves as a neurotransmitter or neuromodulator. Indeed, neuropeptide Y potentiates the vasoconstrictor effect of noradrenaline on blood vessels (7). Also, neuropeptide Y coexists with epinephrine in neurons of locus coeruleus (5), which have been implicated in the central regulation of blood pressure (8). Thus, there is extensive evidence that neuropeptide Y participates in vasoregulation. Neuropeptide Y has also been found to increase food intake (9) and to influence sexual behavior (10) and circadian rhythms (11). The high densities of neuropeptide Y and its receptor in the cerebral cortex and the hippocampus suggest it has a role in higher cognitive functions (12)(13)(14).The amino acid structure of the human neuropeptide Y precursor was deduced from a cDNA clone (15). Human preproneuropeptide Y has 97 amino acid residues and is composed of a signal peptide of 28 amino acids, the mature neuropeptide Y peptide,...

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“…The hybridization cocktail contained 50% formamide, 4 Â SSC (1 Â SSC is, in M, NaCl, 0.15; sodium citrate, 0.015, pH 7.0), 1 Â Denhardt's solution, 1% Sarcosyl, 0.02 M Na 3 PO 4 , pH 7.0, 10% dextransulphate, 0.06 M dithiothreitol, and 0.1 mg/ ml sheared salmon sperm DNA. Single-stranded oligonucleotide 48-mer DNA probes specific for dynorphin (296-345) (Douglass et al, 1989), enkephalin (235-282) (Zurawski et al, 1986), and NPY (1671-1714) (Larhammar et al, 1987) mRNA were used. The probes were 3 0 -end labeled with a-33 P-dATP (Dupont NEN, Wilmington, DE) using terminal deoxynucleotidyl transferase (Gibco) to a specific activity of approximately 1 Â 10 9 c.p.m./mg.…”

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“…CCK (44-mer): 5 Ј -ATCCATCCAGCCCATGTAGTC-CCGGTCACTTATCCTGTGGCTAG-3 Ј according to nucleotides 351-395 of whole rat prepro-cholecystokinin cDNA (Deschenes et al 1984); BDNF (50-mer): 5 Ј -AGTTCCAGTGCCTTTTGTCTATGCCCCTGCAGC-CTTCCTTCGTGTAACCC-3 Ј according to nucleotides 645-694 of whole rat cDNA (Maisonpierre et al 1991); and NPY(48-mer) : 5 Ј -ATGAGATGTGGGGGGAAAC-TAGGAAAAGTCAGGAGAGCAAGTTTCATT-3 Ј according to nucleotides 3148-3195 of rat NPY sequence (Larhammar et al 1987).…”

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