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Characterization of Two Forms of Cocaine- and Amphetamine-Regulated Transcript (CART) Peptide Precursors in Goldfish: Molecular Cloning and Distribution, Modulation of Expression by Nutritional Status, and Interactions with Leptin

Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada

Address all correspondence and requests for reprints to: Dr. R. E. Peter, Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada. E-mail: dick.peter{at}ualberta.ca

Complementary DNAs encoding two forms of cocaine- and amphetamine-regulated transcript (CART) peptide precursors were identified from goldfish brain and named CART I and CART II. Each cDNA contains a signal peptide sequence, the putative CART-like peptide, and a carboxy-terminal extension peptide. Form I encodes a 117-amino acid pro-CART, whereas form II encodes a 120-amino acid pro-CART. Both forms resemble mammalian CART peptides. Each goldfish CART precursor is encoded by three exons interrupted by two introns within genomic DNA. RT-PCR, slot blot, and Northern blot analysis showed that the mRNAs for form I and II precursors have a widespread distribution. Form I and II are present in the brain, pituitary, eye, gonads, and kidney. Form I is also present in the gill. In the brain, form I is predominant in the olfactory bulb and hypothalamus, and form II is predominant in the optic tectum. Food deprivation for 96 h induced a decrease in form I mRNA levels in the telencephalon-preoptic region, hypothalamus, and olfactory bulb and in form II mRNA expression in the olfactory bulb. An increase in mRNA levels was observed 2 h following a meal in the olfactory bulbs and hypothalamus for form I whereas no postprandial changes in form II mRNA levels were observed. Intracerebroventricular injections of human CART alone induced a significant decrease in food intake. Injections of leptin reinforced the inhibition of feeding behavior and food intake seen in CART-treated fish. Central injection of leptin induced an increase in CART I mRNA in the optic tectum, hypothalamus, and olfactory bulbs but had no effect on CART II mRNA expression in the brain. These results suggest that CART peptides act as leptin-regulated satiety factors in goldfish and that they might have other physiological roles besides feeding, possibly in sensory information processing.

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