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Federal government websites often end in. The site is secure. Preview improvements coming to the PMC website in October Learn More or Try it out now. Administration of cocaine and amphetamine increases cocaine- and amphetamine-regulated transcript CART expression in the rat striatum Douglass et al. The presence of CART peptide 55— immunoreactivity in dense core vesicles of axon terminals suggests that the peptide may be released and may act as a neuromodulator Smith et al. Cocaine- and amphetamine-regulated transcript CART peptides are biologically active peptides that mediate feeding and are implicated in psychomotor stimulant drug abuse and stress response Elias et al. CART peptides are processed into shorter biologically active forms from either one or two propeptides in rat Kristensen et al. CART peptide 55— is biologically active and found in the rat brain Kristensen et al. The peptide numbering used here is based on the longer rat propeptide of residues excluding the leader sequence. CART expression increases in the striatum but particularly in the nucleus accumbens within 1 hr after administration of cocaine or amphetamine Douglass et al. CART mRNA and CART peptides are normally expressed throughout the brain, especially in those areas involved in motivation, reward, and feeding, such as the nucleus accumbens, amygdala, and hypothalamus Douglass et al. Consistent with this expression pattern, exogenously applied CART peptides induce many marked behavioral changes, including inhibition of feeding Lambert et al. In addition to the hypothalamus and amygdala, CART peptides are highly expressed in the hippocampus of both rodents and humans Douglass et al. The subcellular localization of CART peptides within dense core vesicles of axon terminals Smith et al. Many neuropeptides, such as neuropeptide Y, which plays a significant role in the regulation of feeding Flynn et al. Rat hippocampal neurons embryonic day 18 isolated as described previously Brewer et al. Fura-2 AM was excited at and nm, and the emission was measured at nm. The background fluorescence at and nm was subtracted from the respective experimental readings, and the ratio of signals at and was obtained. The data were digitized at 2 sec intervals. The data were analyzed using Patch Machine www. After in vitro reduction and refolding, the recombinant CART peptide 55— HIS tag was biologically active, because it reduced food intake in rats. Inhibition of food intake was not seen if the peptide was not refolded, and an improperly folded inactive peptide was used as a control. Other reagents were obtained from Sigma St. Louis, MO. Pretreatment of the neurons with fluorocitrate 0. This suggests that glial cells potentially present in the preparation made an insignificant contribution to the results obtained. The biologically active CART peptide requires appropriate formation of three disulfide bonds Thim et al. Inappropriate formation of these bonds eliminates the ability of CART peptide to inhibit food intake Fritz et al. Reduction of the disulfide links destabilizes the structure, and the peptide loses its ability to inhibit feeding. Recombinant CART peptide 55— that did not undergo the refolding step see Materials and Methods and did not inhibit feeding Fritz et al. Furthermore, we found that the CART action was tissue-specific. The cells were also pretreated with TG. Application of CART peptide 55— to the bath markedly reduced the number of openings elicited by depolarization Fig. Similar results were observed in all seven patches examined Fig. CART peptide 55— applied to the bath produced a dramatic reduction in the number of channel openings recorded in the presence of Bay K Fig. These effects were observed in all five patches examined Fig. The finding that CART peptide 55— added to the bath outside the recording pipette reduced the channel activity suggests that the action of CART peptide 55— is not mediated by the direct blocking or membrane-delimited effect on the channels but is likely to involve diffusible second messengers. CART peptide 55— reduces L-type channel activity. The corresponding ensemble averages are shown at the bottom. The CART peptide application period is indicated by the gray horizontal bar. The mean and median values in the control condition were 0. The ratios of the mean open duration Dur before and after the CART peptide application are also shown using a box plot right. The above data suggest that the CART peptide receptors may be present in rat hippocampal neurons and that diffusible second messengers are likely to mediate signaling of CART peptides. Signal transduction pathways mediating CART peptide 55— action. CART peptide decreases sensitivity of hippocampal neurons to cocaine. Previous studies have elucidated a role for CART peptides in a variety of physiological effects, including inhibition of feeding and increasing anxiety. The cellular mechanism of the CART action was, however, for the most part unknown. Several lines of evidence indicate that this observation is of biological relevance. Rat CART peptide 55— contains three disulfide links to stabilize the biologically active structure Thim et al. Disruption of these disulfide links by reducing agents, such as DTT, is known to destabilize the structure and to render the peptide ineffective in inhibiting feeding Fritz et al. Other neuromodulator peptides have similar effective concentration ranges. Third, the action of CART peptide 55— is tissue-specific, consistent with the idea that some cells contain specific receptors designed for CART peptides. The rat hippocampus may represent a good source of CART peptide receptors for biochemical and molecular studies. If L-type channels are already inhibited by nifedipine, CART peptide 55— should not have any effect. Second, our single-channel recordings show that the channel openings were prolonged by Bay K , and those long openings were in turn inhibited by CART peptide 55— Application of the peptide to the bath inhibited the channel activity recorded within the patch-clamp pipette Fig. The results obtained using pharmacological inhibitors of various intracellular signaling cascades implicate a pertussis toxin-dependent G-protein system in the CART action. The long-lasting nature of the CART action may be surprising, considering the putative G-protein-dependent mechanism. One possible explanation is that CART peptide 55— may be difficult to wash out and that the observed long-lasting effect may not reflect the molecular nature of the underlying intracellular signaling cascade. Our studies with cocaine do offer some clues. The hippocampus has been implicated in the association of environmental cues and the affective states produced by drugs White, ; Vorel et al. Electrical stimulation of the hippocampus elicits cocaine seeking in rats after extinction of cocaine self-administration behavior Vorel et al. Because CART peptides are rapidly produced in response to cocaine application at least in some cells Douglass et al. CART peptides inhibit food intake and may mediate the central action of leptin Elias et al. Our observations further expand the role of CART peptides in the brain, but the exact behaviors affected remain unknown. It is quite possible that CART peptides help shape the neuronal plasticity associated with learning and memory mediated by the hippocampus. We thank Heather Daggett for culturing hippocampal cells. E-mail: gro. As a library, NLM provides access to scientific literature. J Neurosci. Couceyro , 2 and Toshinori Hoshi 1. Pastor R. Abstract Administration of cocaine and amphetamine increases cocaine- and amphetamine-regulated transcript CART expression in the rat striatum Douglass et al. Keywords: hippocampus, neuropeptide, cocaine, CART, calcium, voltage-gated calcium channels. Open in a separate window. Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem. Trends Neurosci. Addiction, dopamine, and the molecular mechanisms of memory. Acute effects of cocaine on human brain activity and emotion. Viable cultured neurons in ambient carbon dioxide and hibernation storage for a month. Optimized survival of hippocampal neurons in Bsupplemented Neurobasal, a new serum-free medium combination. J Neurosci Res. Cocaine: a microstructural analysis of it effects on feeding and associated behavior in the rat. Brain Res. CART peptides: therapeutic potential in obesity and feeding disorders. Drug News Perspect. Further studies on the anatomical distribution of CART by in situ hybridization. J Chem Neuroanat. Receptors couple to L-type calcium channels via distinct G o proteins in rat neuroendocrine cell lines. J Physiol Lond ; — Dolphin AC. Mechanisms of modulation of voltage-dependent calcium channels by G proteins. J Physiol Lond ; :3— L-type calcium channel modulation. Adv Second Messenger Phosphoprotein Res. PCR differential display identifies a rat brain mRNA that is transcriptionally regulated by cocaine and amphetamine. Leptin activates hypothalamic CART neurons projecting to the spinal cord. Farrugia G. G-protein regulation of an L-type calcium channel current in canine jejunal circular smooth muscle. J Membr Biol. Neuropeptide Y-related compounds and feeding. Physiol Behav. Use of fluorocitrate and fluoroacetate in the study of brain metabolism. Generation of CART peptides with in vivo biological activity. Soc Neurosci Abstr. Gawin FH. Cocaine addiction: psychology and neurophysiology. Voltage-independent autocrine modulation of L-type channels mediated by ATP, opioids and catecholamines in rat chromaffin cells. Eur J Neurosci. Different modes of Ca channel gating behaviour favoured by dihyropyridine Ca agonists and antagonists. Hurd YL, Fagergren P. J Comp Neurol. Hyman SE. Addiction to cocaine and amphetamine. Jones SW. Overview of voltage-dependent calcium channels. J Bioenerg Biomembr. Anorexigenic cocaine- and amphetamine-regulated transcript peptide intensifies fear reactions in rats. Immunohistochemical localization of novel CART peptides in rat hypothalamus, pituitary and adrenal gland. J Neuroendocrinol. Cocaine- and amphetamine-regulated transcript peptide immunohistochemical localization in the rat brain. Hypothalamic CART is a new anorectic peptide regulated by leptin. CART peptides: novel addiction- and feeding-related neuropeptides. CART peptide analysis by Western blotting. CART peptides. Regul Pept. A role for novel CART peptide fragments in the central control of food intake. CART peptides in the central control of feeding and interactions with neuropeptide Y. Electrical and calcium signaling in dendrites of hippocampal pyramidal neurons. Annu Rev Physiol. Neuropeptide Y inhibition of calcium channels in PC12 pheochromocytoma cells. Am J Physiol. Miller RJ. Multiple calcium channels and neuronal function. Neurobiological actions of cocaine in the hippocampus. Ann NY Acad Sci. Calcium channel density and hippocampal cell death with age in long-term culture. Premkumar LS. Selective potentiation of L-type calcium channel currents by cocaine in cardiac myocytes. Mol Pharmacol. Staurosporine, K and UCN—potent but nonspecific inhibitors of protein-kinases. Trends Pharmacol Sci. Ultrastructural localization of CART cocaine- and amphetamine-regulated transcript peptides in the nucleus accumbens of monkeys. CART peptide-immunoreactive neurons in the nucleus accumbens in monkeys: ultrastructural analysis, colocalization studies, and synaptic interactions with dopaminergic afferents. CART, a new anorectic peptide. Int J Biochem Cell Biol. Purification and characterisation of a new hypothalamic satiety peptide, cocaine and amphetamine regulated transcript CART , produced in yeast. FEBS Lett. Ui M, Katada T. Bacterial toxins as probe for receptor-G i coupling. Relapse to cocaine-seeking after hippocampal theta burst stimulation. White NM. Addictive drugs as reinforcers: multiple partial actions on memory systems. Chronic cocaine intoxication alters hippocampal sodium channel function. Neurosci Lett. Copy Download.

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