Cocaine- and amphetamine-regulated transcript peptide (CARTp) is an anorexigenic neuropeptide whose receptor is undisclosed. Previously, we reported the specific binding of CART(61-102) to pheochromocytoma PC12 cells, where CART(61-102) affinity and the number of binding sites per cell corresponded to ligand-receptor binding. Recently, Yosten et al. designated orphan GPR160 as the CARTp receptor, because the GPR160 antibody abolished neuropathic pain and anorexigenic effects induced by CART(55-102) and exogenous CART(55-102) coimmunoprecipitated with GPR160 in KATOIII cells. As no direct evidence that CARTp is a ligand for GPR160 has been described, we decided to verify this hypothesis by testing CARTp affinity to the GPR160 receptor. We investigated the GPR160 expression in PC12 cells since it is cell line known to specifically bind CARTp. Moreover, we examined the specific CARTp binding in THP1 cells, with high endogenous GPR160 expression and GPR160-transfected cell lines U2OS and U-251 MG. In PC12 cells, the GPR160 antibody did not compete for specific binding with 125I-CART(61-102) or with 125I-CART(55-102), and GPR160 mRNA expression and GPR160 immunoreactivity were not detected. Moreover, THP1 cells did not show any 125I-CART(61-102) or 125I-CART(55-102) specific binding despite GPR160 detection by fluorescent immunocytochemistry (ICC). Finally, no 125I-CART(61-102) or 125I-CART(55-102) specific binding in the GPR160-transfected cell lines U2OS and U-251 MG, selected due to their negligible endogenous expression of GPR160, was detected, despite the detection of GPR160 by fluorescent ICC. Our binding studies clearly demonstrated that GPR160 cannot be a receptor for CARTp. Further studies are needed to identify true CARTp receptors.

• Klíčová slova
• Binding assay, CARTp, GPCR, GPR160, Transfection,
• MeSH
• kokain * MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• krysa rodu Rattus MeSH
• ligandy MeSH
• proteiny nervové tkáně * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kokain * MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• ligandy MeSH
• proteiny nervové tkáně * MeSH

CART (cocaine- and amphetamine-regulated transcript) peptides are neuropeptides abundant in the central nervous system and periphery found to be involved in the regulation of food intake behavior and other physiological processes. Recently, we reported specific binding of (125)I-CART(61-102) to the rat adrenal pheochromocytoma cell line PC12, both intact cells and cell membranes. In this study, several fragments of CART(61-102) corresponding to its structural loops were synthesized and tested for their potency in binding experiments using PC12 intact cells and cell membranes and in feeding test with fasted mice. From all shorter peptides tested, only CART(74-86) and CART(62-86) containing disulfide bridges kept partial binding potency of the original molecule with K(i) in 10(-5) and 10(-4)M range. However, these fragments were not able to inhibit food intake after their central administration up to a dose of 4 nmol/mouse. The results showed that a compact structure containing three disulfide bridges is necessary for preservation of full biological activity of CART peptides.

• MeSH
• buňky PC12 MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• krysa rodu Rattus MeSH
• molekulární sekvence - údaje MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• peptidové fragmenty farmakologie   MeSH
• proteiny nervové tkáně chemie   farmakologie   MeSH
• sekvence aminokyselin MeSH
• stravovací zvyklosti účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kokainem a amfetaminem regulovaný transkript MeSH
• peptidové fragmenty MeSH
• proteiny nervové tkáně MeSH

CART (cocaine- and amphetamine-regulated transcript) peptides have been studied for ten years. We report specific binding of 125I-CART(61-102) to the rat adrenal pheochromocytoma PC12 cell line, both intact cells and cell membranes. Saturation binding to intact plated cells resulted in Kd of 0.48+/-0.16 nM and Bmax of 2228+/-529 binding sites/cell. 125I-CART(61-102) was also bound to PC12 cells differentiated using nerve growth factor to the neuronal phenotype with non-specific binding below 20%, and Kd of 1.90+/-0.27 nM and Bmax of 11,194+/-261 binding sites/cell. In competitive binding experiments, CART(61-102), CART(55-102) and di-iodinated CART(61-102) were bound to PC12 cell membranes with Ki in low nM range; their affinity to intact non-differentiated and differentiated cells was in low 10(-8) M range. In order to prove that iodination did not eliminate the pharmacological properties of CART, we tested the biological activity of di-iodinated CART(61-102). It decreased food intake in in vivo feeding experiment on fasted mice in a dose of 1 microg/mouse to the same extent as CART(61-102) in a dose of 0.5 microg/mouse.

• MeSH
• biologické modely MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčná membrána metabolismus   MeSH
• buňky PC12 MeSH
• časové faktory MeSH
• fenotyp MeSH
• feochromocytom MeSH
• izotopy jodu metabolismus   MeSH
• kinetika MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• kompetitivní vazba MeSH
• krysa rodu Rattus MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory nadledvin MeSH
• nervový růstový faktor farmakologie   MeSH
• neurony metabolismus   patologie   MeSH
• peptidové fragmenty metabolismus   MeSH
• přijímání potravy účinky léků   MeSH
• proteiny nervové tkáně metabolismus   farmakologie   MeSH
• radioizotopy jodu metabolismus   MeSH
• vazba proteinů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cocaine- and amphetamine-regulated transcript protein (55-102) MeSH Prohlížeč
• izotopy jodu MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• nervový růstový faktor MeSH
• peptidové fragmenty MeSH
• proteiny nervové tkáně MeSH
• radioizotopy jodu MeSH

Monosodium glutamate (MSG) treatment of neonatal mice results in a selective damage to the arcuate nucleus (ARC) and development of obesity with increased adiposity at sustained body weight in the adulthood. Feeding pattern of the MSG obese mice is unusual. Our previous results showed that after 24-h fasting, MSG mice consumed negligible amount of food in several hours and therefore, it was impossible to register the effect of peptides attenuating food intake such as cholecystokinin (CCK) or cocaine- and amphetamine-regulated transcript (CART) peptide. To overcome this problem, two findings were used: firstly, orexigenic effect of neuropeptide Y (NPY) was attenuated both by CCK or CART peptide in lean fed mice and secondly, orexigenic effect of NPY was preserved in fed rats with MSG obesity. In this study, short-term food intake in fed lean and MSG obese C57BL/6 male mice was measured after simultaneous central administration of orexigenic NPY with either CART peptide or peripherally administered CCK. Anorexigenic action of exogenous CART peptide was preserved in MSG obese mice. On the other hand, satiety effect of exogenous CCK was completely lost in MSG obese mice. In conclusion, effective leptin signaling in ARC is necessary for satiety effect of CCK.

• MeSH
• cholecystokinin fyziologie   MeSH
• glutamát sodný MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neuropeptid Y fyziologie   MeSH
• nucleus arcuatus hypothalami fyziologie   MeSH
• obezita chemicky indukované   patofyziologie   MeSH
• proteiny nervové tkáně fyziologie   MeSH
• regulace chuti k jídlu * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cholecystokinin MeSH
• glutamát sodný MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• neuropeptid Y MeSH
• proteiny nervové tkáně MeSH

CART (cocaine- and amphetamine-regulated transcript) peptide is a neuropeptide with a powerful central anorexigenic effect. Specific CART peptide binding sites, most likely CART peptide receptors, have been found in PC12 cells. This study further characterizes the CART peptide binding sites in PC12 cells. After differentiation to a neuronal phenotype with nerve growth factor, the number of CART peptide binding sites in PC12 cells tripled. Following dexamethasone treatment, which transforms PC12 cells into chromaffin-like cells, the number of CART peptide binding sites substantially decreased. CART peptide did not affect the differentiation or acetylcholinesterase activity of PC12 cells, indicating that CART peptide does not participate in differentiation or neuronal activity. CART peptide increased the phosphorylation of SAPK/JNK (stress-activated protein kinase/c-Jun-amino-terminal kinase) and subsequent c-Jun protein expression. These effects were reversed by SP600125, a specific JNK-kinase inhibitor. CART peptide did not significantly affect ERK (extracellular signal-regulated kinase), CREB (cAMP responsive element binding protein), or p38 phosphorylation and c-Fos protein expression. Central administration of CART peptide into mice also resulted in increased c-Jun positive cells in dorsomedial hypothalamic nucleus and nucleus of the solitary tract, areas involved in food intake regulation. Activation of c-Jun by CART peptide might indicate a possible role of CART peptide in managing stress conditions rather than a role in cell proliferation or differentiation as well as the more complex and/or specific regulation ways by transcription factors in some nuclei involved in food intake regulation. The characteristics of stress that CART peptide potentially mediates should be further studied.

• Klíčová slova
• Binding, CART peptide, Differentiation, PC12 cell, Signaling, c-Jun,
• MeSH
• acetylcholinesterasa analýza   MeSH
• buňky PC12 MeSH
• hypothalamus účinky léků   metabolismus   MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• krysa rodu Rattus MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nucleus solitarius účinky léků   metabolismus   MeSH
• proteiny nervové tkáně metabolismus   farmakologie   MeSH
• receptory peptidů metabolismus   MeSH
• signální transdukce fyziologie   MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylcholinesterasa MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• proteiny nervové tkáně MeSH
• receptory peptidů MeSH

BACKGROUND: CART (cocaine- and amphetamine-regulated transcript) peptide and cholecystokinin (CCK) are neuromodulators involved in feeding behavior. This study is based on previously found synergistic effect of leptin and CCK on food intake and our hypothesis on a co-operation of the CART peptide and CCK in food intake regulation and Fos activation in their common targets, the nucleus tractus solitarii of the brainstem (NTS), the paraventricular nucleus (PVN), and the dorsomedial nucleus (DMH) of the hypothalamus. RESULTS: In fasted C57BL/6 mice, the anorexigenic effect of CART(61-102) in the doses of 0.1 or 0.5 microg/mouse was significantly enhanced by low doses of CCK-8 of 0.4 or 4 microg/kg, while 1 mg/kg dose of CCK-A receptor antagonist devazepide blocked the effect of CART(61-102) on food intake. After simultaneous administration of 0.1 microg/mouse CART(61-102) and of 4 microg/kg of CCK-8, the number of Fos-positive neurons in NTS, PVN, and DMH was significantly higher than after administration of each particular peptide. Besides, CART(61-102) and CCK-8 showed an additive effect on inhibition of the locomotor activity of mice in an open field test. CONCLUSION: The synergistic and long-lasting effect of the CART peptide and CCK on food intake and their additive effect on Fos immunoreactivity in their common targets suggest a co-operative action of CART peptide and CCK which could be related to synergistic effect of leptin on CCK satiety.

• MeSH
• antagonisté hormonů farmakologie   MeSH
• benzodiazepinony farmakologie   MeSH
• devazepid farmakologie   MeSH
• fenylmočovinové sloučeniny farmakologie   MeSH
• hubenost * MeSH
• injekce intraperitoneální MeSH
• injekce intraventrikulární MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neurony účinky léků   metabolismus   MeSH
• nucleus dorsomedialis hypothalami účinky léků   fyziologie   MeSH
• nucleus paraventricularis hypothalami účinky léků   fyziologie   MeSH
• nucleus solitarius účinky léků   fyziologie   MeSH
• pátrací chování účinky léků   fyziologie   MeSH
• peptidové fragmenty farmakologie   MeSH
• proteiny nervové tkáně farmakologie   MeSH
• protoonkogenní proteiny c-fos metabolismus   MeSH
• receptor cholecystokininu A antagonisté a inhibitory   MeSH
• receptor cholecystokininu B antagonisté a inhibitory   MeSH
• regulace chuti k jídlu účinky léků   fyziologie   MeSH
• sinkalid farmakologie   MeSH
• synergismus léků MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antagonisté hormonů MeSH
• benzodiazepinony MeSH
• devazepid MeSH
• fenylmočovinové sloučeniny MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• L 365260 MeSH Prohlížeč
• peptidové fragmenty MeSH
• proteiny nervové tkáně MeSH
• protoonkogenní proteiny c-fos MeSH
• receptor cholecystokininu A MeSH
• receptor cholecystokininu B MeSH
• sinkalid MeSH

AIMS: This study investigates the neuroprotective effects of lipidized analogues of 2-SS-CART(61-102) derived from anorexigenic neuropeptide cocaine- and amphetamine-regulated transcript peptide (CARTp) in light of the link between obesity, its comorbidities, and the development of Alzheimer's disease. METHODS: We introduce novel lipidized analogues derived from 2-SS-CART(61-102), a specific analogue of natural CART(61-102), with two disulfide bridges. Using hypothermic PC12 cells, we tested the effect of the most potent analogues on Tau phosphorylation. We further described the anorexigenic and neuroprotective potential of subcutaneously (SC) injected lipidized CARTp analogue in a mouse model with prediabetes and obesity induced by neonatal monosodium glutamate (MSG) administration. RESULTS: Compared to the non-lipidized 2-SS-CART(61-102), all lipidized analogues exhibited a potent binding affinity to PC12 cells and enhanced in vitro stability in rat plasma. Two most potent lipidized analogues attenuated hypothermia-induced Tau hyperphosphorylation at multiple epitopes. Subsequently, chronic SC treatment with palm-2-SS-CART(61-102) significantly decreased body weight and food intake, improved metabolic parameters, decreased level of pTau and increased neurogenesis in hippocampi of obese MSG mice. CONCLUSION: Our unique CARTp analogue palm-2-SS-CART(61-102) shows promise as a potent anti-obesity and neuroprotective agent.

• Klíčová slova
• Alzheimer's disease, CART peptide, Hypothermia, Lipidization, MSG mice, PC12 cells, Tau hyperphosphorylation,
• MeSH
• anorektika farmakologie   MeSH
• buňky PC12 MeSH
• fosforylace účinky léků   MeSH
• glutamát sodný * MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• krysa rodu Rattus MeSH
• lipidy chemie   krev   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neuroprotektivní látky * farmakologie   MeSH
• obezita * metabolismus   farmakoterapie   MeSH
• proteiny nervové tkáně * metabolismus   MeSH
• proteiny tau metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anorektika MeSH
• glutamát sodný * MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• lipidy MeSH
• neuroprotektivní látky * MeSH
• proteiny nervové tkáně * MeSH
• proteiny tau MeSH

CART (cocaine- and amphetamine-regulated transcript) peptides are involved in food intake regulation, stress, and other physiological functions. Although CART peptides have been known for over 25 years, their receptor(s) have not yet been characterized. In this short review, we will summarize our previous studies, where we reported specific binding of 125 I-CART(61-102) to PC12 rat pheochromocytoma cells. Competitive binding experiments performed with mono- and di-iodinated peptides and their isoforms with oxidized Met67 resulted in nanomolar binding affinity. Moreover, in our previous study, CART(61-102), as well as di-iodinated CART(61-102), have shown a strong anorexigenic effect in fasted lean mice after intracerebroventricular administration. In conclusion, from our previous studies, iodination of CART(61-102) resulted in mono- and di-iodinated analogs with or without oxidized Met67 . All analogs revealed a high affinity to binding sites at PC12 cells and preserved biological activity.

• Klíčová slova
• CART peptide, PC12 cells, competitive binding experiments, food intake,
• MeSH
• anorektika chemie   farmakokinetika   terapeutické užití   MeSH
• buňky PC12 MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• krysa rodu Rattus MeSH
• myši MeSH
• proteiny nervové tkáně chemie   farmakokinetika   terapeutické užití   MeSH
• radiofarmaka chemie   farmakokinetika   terapeutické užití   MeSH
• radioizotopy jodu chemie   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• anorektika MeSH
• kokainem a amfetaminem regulovaný transkript MeSH
• proteiny nervové tkáně MeSH
• radiofarmaka MeSH
• radioizotopy jodu MeSH

The CART (cocaine- and amphetamine-regulated transcript) peptide is an anorexigenic neuropeptide that acts in the hypothalamus. The receptor and the mechanism of action of this peptide are still unknown. In our previous study, we showed that the CART peptide binds specifically to PC12 rat pheochromocytoma cells in both the native and differentiated into neuronal phenotype. Two biologically active forms, CART(55-102) and CART(61-102), with equal biological activity, contain three disulfide bridges. To clarify the importance of each of these disulfide bridges in maintaining the biological activity of CART(61-102), an Ala scan at particular S-S bridges forming cysteines was performed, and analogs with only one or two disulfide bridges were synthesized. In this study, a stabilized CART(61-102) analog with norleucine instead of methionine at position 67 was also prepared and was found to bind to PC12 cells with an anorexigenic potency similar to that of CART(61-102). The binding study revealed that out of all analogs tested, [Ala(68,86)]CART(61-102), which contains two disulfide bridges (positions 74-94 and 88-101), preserved a high affinity to both native PC12 cells and those that had been differentiated into neurons. In food intake and behavioral tests with mice after intracerebroventricular administration, this analog showed strong and long-lasting anorexigenic potency. Therefore, the disulfide bridge between cysteines 68 and 86 in CART(61-102) can be omitted without a loss of biological activity, but the preservation of two other disulfide bridges and the full-length peptide are essential for biological activity.

• MeSH
• anorektika chemie   farmakologie   MeSH
• buňky PC12 MeSH
• cystin chemie   MeSH
• kompetitivní vazba MeSH
• krysa rodu Rattus MeSH
• lokomoce účinky léků   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nocicepce účinky léků   MeSH
• peptidové fragmenty chemie   farmakologie   MeSH
• přijímání potravy účinky léků   MeSH
• proteiny nervové tkáně chemie   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• anorektika MeSH
• cocaine- and amphetamine-regulated transcript peptide (62-102), human MeSH Prohlížeč
• cystin MeSH
• peptidové fragmenty MeSH
• proteiny nervové tkáně MeSH

Recent research highlights the profound impact of the gut microbiome on neuropsychiatric disorders, shedding light on its potential role in shaping human behavior. In this study, we investigate the role of the gut microbiome in appetite regulation using activity-based anorexia (ABA) mouse model of anorexia nervosa (AN) - a severe eating disorder with significant health consequences. ABA was induced in conventional, antibiotic-treated, and germ-free mice. Our results show the clear influence of the gut microbiome on the expression of four orexigenic (neuropeptide Y, agouti-related peptide, melanin-concentrating hormone, and orexin) and four anorexigenic peptides (cocaine- and amphetamine-regulated transcript, corticotropin-releasing hormone, thyrotropin-releasing hormone, and pro-opiomelanocortin) in the hypothalamus. Additionally, we assessed alterations in gut barrier permeability. While variations were noted in germ-free mice based on feeding and activity, they were not directly attributable to the gut microbiome. This research emphasizes that the gut microbiome is a pivotal factor in AN's appetite regulation beyond just dietary habits or physical activity.

• Klíčová slova
• Activity-based anorexia model, Appetite regulation, Eating disorders, Gut barrier, Microbiome, Neuropeptide Y,
• MeSH
• chuť k jídlu fyziologie   MeSH
• hypothalamus metabolismus   MeSH
• lidé MeSH
• mentální anorexie * metabolismus   MeSH
• myši MeSH
• neuropeptidy * metabolismus   MeSH
• střevní mikroflóra * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• neuropeptidy * MeSH