中华鲟(Acipenser sinensis)脑垂体的形态和ACTH细胞定位
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摘要
为研究中华鲟(Acipenser sinensis)神经内分泌系统的基本结构和功能,本文对中华鲟脑垂体的组织形态进行了观察,结果表明中华鲟的神经垂体较小,腺垂体可分为前、中和后腺垂体三部分,神经垂体形成分支插入后腺垂体。中华鲟垂体还有一个特点是有分支的垂体裂。前腺垂体位于垂体前侧,细胞间隙大,细胞分散呈窦状。中腺垂体细胞间隙小,细胞密集呈条索状,嗜酸性细胞和嗜碱性细胞区分明显。后腺垂体包围在神经垂体周围。腺垂体中有长且分支的垂体裂,腺垂体形成许多突起深入其中,垂体裂周围的细胞密集,染色深,排列成类似上皮组织的形态。免疫组化结果表明,中华鲟脑垂体ACTH细胞主要分布于后腺垂体。ACTH的前体基因是POMC,RT-PCR证明其主要分布在脑垂体中,在体内其他组织中基本检测不到。编码ATCH的片段位于POMC中最保守的区域。预测发现POMC基因有一个26氨基酸的信号肽,将不带信号肽的POMC基因插入表达载体。再转染HELA细胞POMC主要在细胞质中表达,而带有信号肽的POMC蛋白在细胞内很少,推测可能分泌到胞外,信号肽对POMC表达和功能发挥起重要作用,本研究结果可为该基因在中华鲟神经内分泌研究中提供参考。
To understand the structure and function of the neuroendocrine system of Chinese sturgeon Acipenser sinensis, we studied the morphology of the pituitary and the adrenocorticotropin(ACTH) cells in the pituitary gland. The fish were sampled from Changjiang(Yangtze) River estuary, body weight ~100 g. We observed that the fish has a small neurohypophysis(Ne), and adenohypophysis could be divided into three parts: proadenophypophysis(Pa), mesoadenohypophysis(Ma) and metaadenohypophysis(Me). Pa is located in the front of the pituitary. The Ne is small and finger-like in the middle part, not in the Pa as most teleost fish. Another feature is the presence of a large hypophysial cleft(HC) with numerous digitations. The cells of adenohypophysis contained the basophil cells, acidophil cells, and chromophobe cells, and they were different in shape and in distribution. The cells around the HC looked like epithelial lined. Immunohistochemical study showed that the ACTH cells in the pituitary gland distributed mainly in the Me. The precursor gene of ACTH is POMC(pro-opiomelanocortin), and RT-PCR shows that it distributed in the pituitary but other tissues. The ATCH-encoding region was located in the most conservative region of the POMC. It is predicted that the POMC gene has a 26-amino-acid signal peptide. The POMC gene with or without signal peptide was cloned into the expression vector. After transfection into He La cells, POMC without signal peptide was expressed in the cytoplasm, while the POMC protein with signal peptide was very little in the cytoplasm. Proteins with a signal peptide are rare in cells and they probably secreted out of the cells. Therefore, signal peptide plays an important role in the expression and function of POMC. The findings shall provide a reference for the study of Chinese sturgeon neuroendocrine.
To understand the structure and function of the neuroendocrine system of Chinese sturgeon Acipenser sinensis, we studied the morphology of the pituitary and the adrenocorticotropin(ACTH) cells in the pituitary gland. The fish were sampled from Changjiang(Yangtze) River estuary, body weight ~100 g. We observed that the fish has a small neurohypophysis(Ne), and adenohypophysis could be divided into three parts: proadenophypophysis(Pa), mesoadenohypophysis(Ma) and metaadenohypophysis(Me). Pa is located in the front of the pituitary. The Ne is small and finger-like in the middle part, not in the Pa as most teleost fish. Another feature is the presence of a large hypophysial cleft(HC) with numerous digitations. The cells of adenohypophysis contained the basophil cells, acidophil cells, and chromophobe cells, and they were different in shape and in distribution. The cells around the HC looked like epithelial lined. Immunohistochemical study showed that the ACTH cells in the pituitary gland distributed mainly in the Me. The precursor gene of ACTH is POMC(pro-opiomelanocortin), and RT-PCR shows that it distributed in the pituitary but other tissues. The ATCH-encoding region was located in the most conservative region of the POMC. It is predicted that the POMC gene has a 26-amino-acid signal peptide. The POMC gene with or without signal peptide was cloned into the expression vector. After transfection into He La cells, POMC without signal peptide was expressed in the cytoplasm, while the POMC protein with signal peptide was very little in the cytoplasm. Proteins with a signal peptide are rare in cells and they probably secreted out of the cells. Therefore, signal peptide plays an important role in the expression and function of POMC. The findings shall provide a reference for the study of Chinese sturgeon neuroendocrine.
引文
刘婷,章龙珍,张涛等,2011.中华鲟神经内分泌多肽7B2基因克隆及组织特异性表达.海洋渔业,33(2):121-130
张涛,庄平,章龙珍等,2011.长江口中华鲟自然保护区底层鱼类的群落结构特征.生态学报,31(6):1687-1694
陈秋生,2002.兽医比较组织学.北京:中国农业出版社
陈晓武,刘仪,毕燕会等,2010.中华鳖脑垂体的显微与超微结构观察.水产学报,34(8):1210-1218
班璇,肖飞,2014.葛洲坝下游河势调整工程对中华鲟产卵场的影响.水利学报,45(1):58-64
曹宏,李创举,危起伟等,2011.中华鲟阿黑皮素原c DNA序列克隆及其序列分析.水生生物学报,35(5):776-782
楼允东,1980.组织胚胎学.2版.北京:中国农业出版社
Alrubaian J,Danielson P,Fitzpatrick M et al,1999.Cloning of a second proopiomelanocortin c DNA from the pituitary of the sturgeon,Acipenser transmontanus.Peptides,20(4):431-436
Alrubaian J,Sollars C,Danielson P B et al,2003.Evaluating the radiation of the POMC gene in teleosts:characterization of American eel POMC.General and Comparative Endocrinology,132(3):384-390
Amemiya Y,Takahashi A,Suzuki N et al,1999.A newly characterized melanotropin in proopiomelanocortin in pituitaries of an elasmobranch,Squalus acanthias.General and Comparative Endocrinology,114(3):387-395
Arends R J,Vermeer H,Martens G J et al,1998.Cloning and expression of two proopiomelanocortin m RNAs in the common carp(Cyprinus carpio L.).Molecular and Cellular Endocrinology,143(1-2):23-31
Danielson P B,Alrubaian J,Muller M et al,1999.Duplication of the POMC gene in the paddlefish(Polyodon spathula):analysis of gamma-MSH,ACTH,and beta-endorphin regions of ray-finned fish POMC.General and Comparative Endocrinology,116(2):164-177
Ferrandino I,Pica A,Grimaldi M C,2000.Immunohistochemical detection of ACTH and MSH cells in the hypophysis of the hermaphroditic teleost,Diplodus sargus.European Journal of Histochemistry Ejh,44(4):397-406
Follenius E,Dubois M P,1976.Cytology and immunocytology of the organization of the intermediate lobe of the perch(Perca fluviatilis L.)pituitary.General&Comparative Endocrinology,30(4):462-471
Goos Th H J,1982.Proceedings of the International Symposium on Reproductive Physiology of Fish.Wageningen:Centre for Agricultural Pub.and Documentation
Gorbman A,1995.Olfactory origins and evolution of the brain-pituitary endocrine system:facts and speculation.General and Comparative Endocrinology,97(2):171-178
Harris R M,Dijkstra P D,Hofmann H A,2014.Complex structural and regulatory evolution of the pro-opiomelanocortin gene family.General and Comparative Endocrinology,195:107-115
Heinig J A,Keeley F W,Robson P et al,1995.The appearance of proopiomelanocortin early in vertebrate evolution:cloning and sequencing of POMC from a Lamprey pituitary c DNAlibrary.General and Comparative Endocrinology,99(2):137-144
Howe A,1973.The mammalian pars intermedia:a review of its structure and function.Journal of Endocrinology,59(2):385-409
Kawauchi H,1989.Evolutionary aspects of pituitary hormones.Kitasato Archives of Experimental Medicine,62(4):139-155
Kitahara N,Nishizawa T,Iida K et al,1988.Absence of aγ-melanocyte-stimulating hormone sequence in proopiomelanocortin m RNA of chum salmon Oncorhynchus keta.Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,91(2):365-370
Kuhnen P,Clément K,Wiegand S et al,2016.Proopiomelanocortin deficiency treated with a melanocortin-4 receptor agonist.New England Journal of Medicine,375(3):240-246
Lee J,Danielson P,Sollars C et al,1999.Cloning of a neoteleost(Oreochromis mossambicus)pro-opiomelanocortin(POMC)c DNA reveals a deletion of the gamma-melanotropin region and most of the joining peptide region:implications for POMC processing.Peptides,20(12):1391-1399
Mencarelli M,Zulian A,Cancello R et al,2012.A novel missense mutation in the signal peptide of the human POMCgene:a possible additional link between early-onset type 2diabetes and obesity.European Journal of Human Genetics,20(12):1290-1294
Miraglia D G E,Cirillo G,Santoro N et al,2001.Molecular screening of the proopiomelanocortin(POMC)gene in Italian obese children:report of three new mutations.International Journal of Obesity,25(1):61-67
Pelissero C,Nunez-Rodriguez J,Le Menn F et al,1988.Immunohistochemical investigation of the pituitary of the sturgeon(Acipenser baeri,Chondrostei).Fish Physiology and Biochemistry,5(3):109-119
Prakash A,2015.Studies on the functional morphology of the endocrine glands in some fresh water fishes.Cybernetics&Systems,38(7):667-693
Pritchard L E,Turnbull A V,White A,2002.Pro-opiomelanocortin processing in the hypothalamus:impact on melanocortin signalling and obesity.Journal of Endocrinology,172(3):411-421
Raffan E,Dennis R J,O'Donovan C J et al,2016.A deletion in the canine POMC gene is associated with weight and appetite in obesity-prone Labrador retriever dogs.Cell Metabolism,23(5):893-900
Suzuki M,Bennett P,Levy A et al,1997.Expression of MCH and POMC genes in rainbow trout(Oncorhynchus mykiss)during ontogeny and in response to early physiological challenges.General and Comparative Endocrinology,107(3):341-350
Vallarino M,Delbende C,Bunel D T et al,1989.Proopiomelanocortin(POMC)-related peptides in the brain of the rainbow trout,Salmo gairdneri.Peptides,10(6):1223-1230
Varsamos S,Bonga B S W,Flik G et al,2003.Cloning of a proopiomelanocortin c DNA from the pituitary gland of the sea bass(Dicentrarchus labrax)and assessment of m RNAexpression in different tissues by means of real-time PCR.Journal of Endocrinology,176(3):405-414
Wallis M,1975.The molecular evolution of pituitary hormones.Biological Reviews,50(1):35-98
张涛,庄平,章龙珍等,2011.长江口中华鲟自然保护区底层鱼类的群落结构特征.生态学报,31(6):1687-1694
陈秋生,2002.兽医比较组织学.北京:中国农业出版社
陈晓武,刘仪,毕燕会等,2010.中华鳖脑垂体的显微与超微结构观察.水产学报,34(8):1210-1218
班璇,肖飞,2014.葛洲坝下游河势调整工程对中华鲟产卵场的影响.水利学报,45(1):58-64
曹宏,李创举,危起伟等,2011.中华鲟阿黑皮素原c DNA序列克隆及其序列分析.水生生物学报,35(5):776-782
楼允东,1980.组织胚胎学.2版.北京:中国农业出版社
Alrubaian J,Danielson P,Fitzpatrick M et al,1999.Cloning of a second proopiomelanocortin c DNA from the pituitary of the sturgeon,Acipenser transmontanus.Peptides,20(4):431-436
Alrubaian J,Sollars C,Danielson P B et al,2003.Evaluating the radiation of the POMC gene in teleosts:characterization of American eel POMC.General and Comparative Endocrinology,132(3):384-390
Amemiya Y,Takahashi A,Suzuki N et al,1999.A newly characterized melanotropin in proopiomelanocortin in pituitaries of an elasmobranch,Squalus acanthias.General and Comparative Endocrinology,114(3):387-395
Arends R J,Vermeer H,Martens G J et al,1998.Cloning and expression of two proopiomelanocortin m RNAs in the common carp(Cyprinus carpio L.).Molecular and Cellular Endocrinology,143(1-2):23-31
Danielson P B,Alrubaian J,Muller M et al,1999.Duplication of the POMC gene in the paddlefish(Polyodon spathula):analysis of gamma-MSH,ACTH,and beta-endorphin regions of ray-finned fish POMC.General and Comparative Endocrinology,116(2):164-177
Ferrandino I,Pica A,Grimaldi M C,2000.Immunohistochemical detection of ACTH and MSH cells in the hypophysis of the hermaphroditic teleost,Diplodus sargus.European Journal of Histochemistry Ejh,44(4):397-406
Follenius E,Dubois M P,1976.Cytology and immunocytology of the organization of the intermediate lobe of the perch(Perca fluviatilis L.)pituitary.General&Comparative Endocrinology,30(4):462-471
Goos Th H J,1982.Proceedings of the International Symposium on Reproductive Physiology of Fish.Wageningen:Centre for Agricultural Pub.and Documentation
Gorbman A,1995.Olfactory origins and evolution of the brain-pituitary endocrine system:facts and speculation.General and Comparative Endocrinology,97(2):171-178
Harris R M,Dijkstra P D,Hofmann H A,2014.Complex structural and regulatory evolution of the pro-opiomelanocortin gene family.General and Comparative Endocrinology,195:107-115
Heinig J A,Keeley F W,Robson P et al,1995.The appearance of proopiomelanocortin early in vertebrate evolution:cloning and sequencing of POMC from a Lamprey pituitary c DNAlibrary.General and Comparative Endocrinology,99(2):137-144
Howe A,1973.The mammalian pars intermedia:a review of its structure and function.Journal of Endocrinology,59(2):385-409
Kawauchi H,1989.Evolutionary aspects of pituitary hormones.Kitasato Archives of Experimental Medicine,62(4):139-155
Kitahara N,Nishizawa T,Iida K et al,1988.Absence of aγ-melanocyte-stimulating hormone sequence in proopiomelanocortin m RNA of chum salmon Oncorhynchus keta.Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,91(2):365-370
Kuhnen P,Clément K,Wiegand S et al,2016.Proopiomelanocortin deficiency treated with a melanocortin-4 receptor agonist.New England Journal of Medicine,375(3):240-246
Lee J,Danielson P,Sollars C et al,1999.Cloning of a neoteleost(Oreochromis mossambicus)pro-opiomelanocortin(POMC)c DNA reveals a deletion of the gamma-melanotropin region and most of the joining peptide region:implications for POMC processing.Peptides,20(12):1391-1399
Mencarelli M,Zulian A,Cancello R et al,2012.A novel missense mutation in the signal peptide of the human POMCgene:a possible additional link between early-onset type 2diabetes and obesity.European Journal of Human Genetics,20(12):1290-1294
Miraglia D G E,Cirillo G,Santoro N et al,2001.Molecular screening of the proopiomelanocortin(POMC)gene in Italian obese children:report of three new mutations.International Journal of Obesity,25(1):61-67
Pelissero C,Nunez-Rodriguez J,Le Menn F et al,1988.Immunohistochemical investigation of the pituitary of the sturgeon(Acipenser baeri,Chondrostei).Fish Physiology and Biochemistry,5(3):109-119
Prakash A,2015.Studies on the functional morphology of the endocrine glands in some fresh water fishes.Cybernetics&Systems,38(7):667-693
Pritchard L E,Turnbull A V,White A,2002.Pro-opiomelanocortin processing in the hypothalamus:impact on melanocortin signalling and obesity.Journal of Endocrinology,172(3):411-421
Raffan E,Dennis R J,O'Donovan C J et al,2016.A deletion in the canine POMC gene is associated with weight and appetite in obesity-prone Labrador retriever dogs.Cell Metabolism,23(5):893-900
Suzuki M,Bennett P,Levy A et al,1997.Expression of MCH and POMC genes in rainbow trout(Oncorhynchus mykiss)during ontogeny and in response to early physiological challenges.General and Comparative Endocrinology,107(3):341-350
Vallarino M,Delbende C,Bunel D T et al,1989.Proopiomelanocortin(POMC)-related peptides in the brain of the rainbow trout,Salmo gairdneri.Peptides,10(6):1223-1230
Varsamos S,Bonga B S W,Flik G et al,2003.Cloning of a proopiomelanocortin c DNA from the pituitary gland of the sea bass(Dicentrarchus labrax)and assessment of m RNAexpression in different tissues by means of real-time PCR.Journal of Endocrinology,176(3):405-414
Wallis M,1975.The molecular evolution of pituitary hormones.Biological Reviews,50(1):35-98