2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38544CART (cocaine- and amphetamine-regulated transcript) is an anorexigenic neurotransmitter that can be induced by cocaine or amphetamine (AMPH). Daily treatment of AMPH, an anorectic agent, for several days induced a marked anorexia (AMPH anorexia) on day 1 and a gradual tolerance to this anorexia (AMPH tolerance) on the followings. Both AMPH anorexia and tolerance are relevant to the alteration of neuropeptide Y (NPY), an orexigenic agent abundant in the brain. The mechanism for the anorectic effect of AMPH is due to the central release of dopamine which exerts its inhibitory action on NPY neurons. Recently, we found that CART might mediate the action of dopamine on NPY neurons. Aim of the present study is to assess this hypothesis. Contents of the present study can be divided into three Parts that will be completed within 3 years. The protocol title for Part 1 is to investigate the role of CART in the mediation of AMPH-induced anorexia, for Part 2 is to examine the role of CART in the induction of AMPH neurotoxicity and for Part 3 is to examine the role of PKA/CREB signaling in the regulation of CART-mediated anorexia. There are three experiments that need to be examined for each Part of investigation. In Part 1, it included experiments that examine: (1) the effect of AMPH treatment on CART and NPY gene expression, (2) the effect of AMPH treatment on CART and NPY immunoreactivity, and (3) the effect of ICV injection of CART antisens on AMPH-induced anorexia. In Part 2, it included experiments that examine: (1) the effect of AMPH neurotoxicity on CART, superoxide dismutase (SOD), nitric oxide synthase (NOS) and NPY gene expression, (2) the effect of CART antisense pretreatment on AMPH neurotoxicity, and (3) the effect of free radical inhibitor on CART gene expression. In Part 3, it included experiments that examine: (1) the effect of AMPH treatment on CART, PKA, CREB and NPY gene expression, (2) the effect of AMPH treatment on CART, SOD and NOD immunoreactivity, and (3) the effect of ICV injection of PKA or CREB antisense on CART-mediated anorexia. Moreover, in each Part of the investigation, there are two sets of animals, i.e., AMPH anorexia and AMPH tolerance groups, which will be used to study the acute and chronic treatment of AMPH, respectively.CART (cocaine- and amphetamine-regulated transcript) peptide 是一種厭食性神經傳遞物，可由安非他命(amphetamine, AMPH)誘導產生。Y型神經勝.(neuropeptide Y, NPY)是腦內含量豐富可興奮食慾的神經傳遞物。AMPH若每天注射一次，持續數天時，在第一天可引發最顯著的厭食效應(本文稱為AMPH anorexia)，但在隨後數天，則逐漸產生厭食耐受性(本文稱為AMPH tolerance)。AMPH anorexia與「抑制NPY基因的表現」有關，而AMPH tolerance則與「NPY基因表現的逐漸回升」有關。安非他命厭食作用的機制，是透過腦內dopamine的釋放，去影響腦內NPY基因表現所致。最近，我們發現此作並非直接的，可能透過CART媒介此厭食作用，因此，本計畫將探討此假說。計畫將分成三個子計畫，擬分三年分別探討，子計畫名稱為：(1)下視丘中CART在安非他命誘發之厭食效應之角色，(2)CART在安非他命誘發之毒性之角色,(3)PKA/CREB訊息傳遞在CART媒介之厭食效應之角色。此外，每個子計畫的實驗將分「AMPH anorexia」及「AMPH tolerance」兩組動物，分別探討急性安非他命及慢性安非他命的作用。 第一年預期完成三項工作：(1) 安非他命注射對CART、NPY基因表現的影響，(2)以「雙重免疫顆粒染色法」觀察CART及NPY免疫顆粒之表現，(3)腦室注射CART antisense oligodeoxynucleotide對安非他命厭食行為的影響。第二年預期完成三項工作：(1) 安非他命神經毒性(AMPH neurotoxicity) 對CART, superoxide dismutase (SOD), nitric oxide synthase (NOS)及NPY基因表現的影響，(2)腦室注射CART antisense對AMPH neurotoxicity的影響，(3)自由基抑制劑 (melatonin或7-nitroindazole) 前處理對CART基因表現的影響。第三年預期完成三項工作：(1) 安非他命對CART、PKA、CREB、c-fos及c-jun基因表現的影響，(2)以「雙重免疫顆粒染色法」觀察CART及PKA (or CREB)免疫顆粒之表現，(3)下視丘旁室核注射PKA或CREB antisense對CART厭食行為的影響。古柯鹼和安非他命調控之轉錄因子(CART)安非他命毒性攝食行為Y型神經胜(NPY)訊息傳遞蛋白激A(PKA)cAMP反應因子結合蛋白(CREB)CARTamphetamine neurotoxicityneuropeptide Y (NPY)signal transductionPKA (protein kinase A)CREB (cAMP response element binding protein)food intakeSODNOSantisense oligodeoxynucleotideProject