博碩士論文 88224010 詳細資訊




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姓名 龎慶弘(Chin-Hon Pan)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 中華鱉腦垂體甘丙氨激素之研究:cDNA選殖、表現及調控
(Studies on pituitary galanin of Chinese mud turtle: cDNA clonig, expression and regulation)
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摘要(中) 甘丙胺激素是一種C端有乙醯化的神經內泌素,已知其具有調控脊椎動物行為與食慾的功能。為了對爬行類腦垂體甘丙胺激素的物種歧異度與基因之演化有更進一步之了解,本研究以中華鱉為實驗動物,利用快速核酸兩端放大之技術,定序腦垂體甘丙胺激素互補核苷酸序列,所得的序列共有672個核苷酸,5’-未轉錄區99 個;前原甘丙胺激素有324個核苷酸;3’-未轉錄區有249 個。前原甘丙胺激素有108個胺基酸;本體為29個胺基酸,與其他脊椎動物之間的同源性為72% ~ 96%。 半定量聚合酶連鎖反應實驗結果顯示,前原甘丙胺激素於腦、下丘腦、胃、肝臟、胰臟、睪丸、卵、子宮、大腸與小腸皆有表現,而在心臟、脾臟、肌肉與脂肪組織則無法偵測到其表現。雌性前原甘丙胺激素之表現量似乎高於雄性。 離體腦垂體組織培養研究顯示:雌二醇(10-6M~10-8M)、性釋素(10-7M~10-9M) 隨著濃度增加會明顯促進前原甘丙胺激素之表現;而瘦素(10-6M~10-8M)與神經月太 Y(10-6M~10-8M)隨著濃度增加,抑制甘丙胺激素基因表現之情形更趨明顯。 綜觀上述結果顯示脊椎動物之前原甘丙胺激素具高度之保守性,且與雌二醇、性釋素、瘦素及神經月太 Y在中華鱉腦垂體的生理功能上具交互作用。
摘要(英) Galanin is a C-terminally-amidated neurohormone known to regulate various aspects of behavior and appetite of vertebrates. To better understanding the phylogenetic diversity and evolution of galanin genes in reptilie, we here conducted an analysis of galanin cDNA from Chinese mud turtle pituitary. Using 3’- and 5’- RACE techniques, we have isolated and sequenced a novel cDNA derived from pituitary total RNA. Sequence analysis revealed a 672-bp cDNA containing a 99-bp 5’-untranslated region, a 324-bp preprogalanin coding region, and a 249-bp 3’-untranslated region. The deduced protein is a 108-amino acid preprogalanin molecule with [Ser23]-galanin-29 at the C-terminus of the coding region, making its homology to other vertebrates in the range of 72%~96%. Semiquantitative PCR revealed that preprogalanin was expressed in the brain, hypothalamus, pituitary, stomach, liver, pancreas, testis, ovary, uterus, small intestine and large intestine, but undetected in the heart, spleen, muscle and adipose tissue. Females seemed to express more preprogalanin than males. In vitro pituitary tissue culture study indicated that E2 (10-6M~10-8M) or GnRH (10-7M~10-9M) treatment significantly increased the levels of preprogalaninin a dose- dependent manner. However, Lep (10-6M~10-8M) or NPY (10-7M~10-9M) treatment decreased the levels of preprogalanin also in a dose-dependent manner. These results suggest that preprogalanin is conserved in vertebrates and that a physiological role of galanin interacting with E2, GnRH, Lep and/or NPY may exist in the pituitary of mud turtle.
關鍵字(中) ★ 中華鱉
★ 甘丙胺激素
關鍵字(英) ★ galanin
★ Chinese mud turtle
論文目次 目錄
中文摘要 1
英文摘要 2
縮寫與全名對照 3
第一章:簡介 4
壹、中華鱉 4
貳、甘丙胺激素 4
1. 歷史 4
2. 種類 4
3. 結構 4
4. 基因序列 4
5. 組織分佈與血中含量 5
6. 接受器 5
7. 生物功能 6
8. 內分泌調控甘丙胺激素之表現 6
參、研究目的 7
第二章:材料與方法 8
壹、實驗材料 8
貳、實驗動物 8
參、實驗方法 8
一、快速核酸兩端放大 (RACE) 8
1. RNA萃取 (RNA Extraction) 9
2. 3´RACE 9
A. 第一股cDNA合成 9
B. 聚合酶連鎖反應(PCR) 10
3. 5´RACE 11
A. 第一股cDNA合成 11
B. 聚合酶連鎖反應(PCR) 11
二、組織特異性實驗 11
1. RNA萃取 (RNA Extraction) 11
A. 第一股cDNA合成 12
B. 聚合酶連鎖反應(PCR) 12
2. 第一股cDNA合成 12
3. 聚合酶連鎖反應(PCR) 12
三、激素對甘丙胺激素表達情形影響之實驗 13
1. 鱉腦垂體於M199離體培養實驗 13
2. 激素處理 13
第三章:結果 14
壹、中華鱉腦垂體甘丙胺激素cDNA序列 14
貳、組織特異性 14
參、激素對甘丙胺激素基因表達之影響 14
1. 甘丙胺激素基因於M199離體培養之表現情形 14
2. 雌二醇促進甘丙胺激素基因的表現 15
3. 瘦素抑制甘丙胺激素基因的表現 15
4. 雌二醇與瘦素之交互影響甘丙胺激素基因的表現 15
5. 性釋素促進甘丙胺激素基因的表現 16
7. 神經月太 Y抑制甘丙胺激素基因的表現 16
8. 性釋素與神經月太 Y之交互影響甘丙胺激素基因的表現 17
第四章:討論 18
壹、中華鱉腦垂體甘丙胺激素cDNA序列 18
貳、組織特異性 19
參、激素影響中華鱉腦垂體甘丙胺激素基因的表現 19
第五章:結論與展望 20
第六章:參考文獻 21
第七章:附錄 27
圖目錄 Ⅳ
表目錄 Ⅵ
圖目錄
圖一、 哺乳動物甘丙胺激素的基因表現受到其他激素的調控 28
圖二、 本研究所採用之引子於甘丙胺激素的相對位置 29
圖三、 鱉腦垂體甘丙胺激素cDNA序列 30
圖四、 中華鱉甘丙胺激素的組織特異性 31
圖五、 離體培養中華鱉腦垂體甘丙胺激素基因表現之時間型態 32
圖六、 雌二醇 (10-6M) 對中華鱉腦垂體甘丙胺激素mRNA表現的影響 33
圖七、 雌二醇 (10-8 M) 對中華鱉腦垂體甘丙胺激素mRNA表現的影響 34
圖八、 瘦素對中華鱉腦垂體甘丙胺激素mRNA表現的影響 35
圖九、 雌二醇 (10-8M) 與瘦素 (10-8M) 交互處理6小時對中華鱉腦垂體甘丙胺激素mRNA表現的影響 36
圖十、 雌二醇 (10-8M) 與瘦素 (10-8M) 交互處理12小時對中華鱉腦垂體甘丙胺激素基因表現的影響 37
圖十一、 性釋素 (GnRH) 對中華鱉腦垂體甘丙胺激素mRNA表現的影響 38
圖十二、性釋素 (GnRH, 10-9 M) 對中華鱉腦垂體中甘丙胺激素mRNA表現的影響 39圖十三、 神經月太 Y (NPY) 對中華鱉腦垂體甘丙胺激素mRNA表現的影響 40
圖十四、 性釋素 (GnRH,10-9 M) 與神經月太 Y (NPY, 10-9 M) 交互處理12小時對中華鱉腦垂體甘丙胺激素mRNA表現的影響 41
圖十五 、脊椎動物甘丙胺激素之演化樹 (phylogenic tree) 42
圖十六、 激素對中華鱉腦垂體甘丙胺激素的調控 43
表目錄
表一 :甘丙胺激素對脊椎動物荷爾蒙的影響 44
表二 :本研究所採用之引子 (primer) 序列 45表三 :中華鱉之前原甘丙胺激素(preprogalanin) cDNA序列與其他脊椎動物之同源性 46
表四 :中華鱉甘丙胺激素胺基酸序列與脊椎動物之同源性 47
表五 :脊椎動物甘丙胺激素的胺基酸序列 48
參考文獻 Argiolas A. (1999). Neuropeptides and sexual behavior. Neurosci Biobehav Rev, 23, 1127-1142.
Baranowska B, Chemielowska M, Radzikowska M, Borowiec M, Roguski K, Wasilewski-Dziubinska E. (1999). Effects of neuropeptide Y (NPY), galanin and vasoactiveintestinal peptide (VIP) on pituitary hormone release and on ovarian steroidogenesis. Neuroendocrinol Lett, 20, 385-389.
Beck B, Stricker-Krongard A, Burlet A, Cumin F, Burlet C. (2001). Plasma leptin and hypothalamic neuropeptide Y and galanin levels in long-evans rats with marked dietary preferences. Nutr Neurosci, 4, 39-50.
Bersani M, Johnsen AH, Højrup P, Dunning BE, Andreasen JJ, Holst JJ. (1991). Human galanin: primary structure and identification of two molecular forms. FEBS Letters, 283, 189-194.
Bloch GJ, Butler PC, Kohlert JG, Bloch DA. (1996). Galanin microinjected into the medial preoptic nucleus facilitates female-and-male typical sexual behavior in the female rat. Physol Behav, 59, 1147-1154.
Bloch GJ, Butler PC, Kohlert JG, Bloch DA. (1993). Microinjection of galanin into the medial preoptic nucleus facilitates copulatory behavior in the male rat. Physol Behav, 54, 615-624.
Cheung CC, Hohmann JG, Clifton DK, Steiner RA. (2001). Distribution of galanin messenger rna-expressing cells in murine brain and their regulation by leptin in regions of the hypothalamus. Neuroscience, 103, 423-432.
Cimini V. (2000). Galanin coexists with pituitary hormones in the fetal rat. Peptides, 21, 1711-1716.
Crawley JN*. (1995). Biological action of galanin. Regulatory peptides, 59, 1-16.
Crawley JN. (1999). The role of galanin in feeding behavior. Neuropepetides, 33, 369-375.
Delemarre-van de waal HA, Burton KA, Kabigting EB, Steiner RA, Clifton DK. (1994). Expression and sexual dimorphisom of galanin messenger ribonucleic acid in growth hormone-releasing hormone neurons of the rat during development. Endocrinology, 134, 665-671.
Evans H, Baumgartner M, Shine J, Herzog H. (1993). Genomic organization and localization of the gene encoding human preprogalanin. Genomics, 18, 473-477.
Elsaesser F. (1995). Stimulation of porcine pituitary luteinizing hormone release by galanin: putative auto/paracrine regulation. Neuroendocrinology, 74, 288-299.
Finn PD, McFall TB, Clifton DK, Steiner RA. (1996). Sexual differentiation of galanin gene expression in gonadotropin-releasing hormone neurons. Endocrinology, 137, 4767-4772.
Genazzani AR, Bernardi F, Monteleone P, Luisi S, Luisi M. (2000). Neuropeptides, Neurotransmitters neurosteroids, and the onset of puberty. Ann N Y Acad Sci, 900, 1-9.
Guistina A, Licini M, Bussi AR, Girelli A, Pizzocolo G, Schettino M, Negro-Vilar A. (1993). Effects of sex and age on the growth hormone response to galanin in healthy human subjects. J Clin Endocrinol Metab, 76, 1369-1372.
Hsieh YL, Chatterjee A, Chien JT, Yu JYL. (2001). Molecular cloning of the cDNAs for pituitary glycoprotein hormone α-subunit of two species of duck and their gene regulation. JME, 27, 339-347.
Hsieh YL, Chatterjee A, Lee G, Yu JYL. (2000). Molecular cloning and sequence analysis of the cDNA for thyroid-stimulating hormone β-subunit of muscovy duck. GCE, 120, 336-344.
Ingvartsen KL, Boisclair YR. (2001). Leptin and the regulation of food intake, energy homeostasis and immunity with special focus on periparturient ruminants. Domest Anim Endocrinol, 21, 215-250.
Karelson E, Langel U. (1998). Galaninergic signalling and adenylate cyclase. Neuropepetides, 32,197-210.
Kofler B, Liu ML, Jacoby AS, Shine J, Iismaa TP. (1996). Molecular cloning and characterization of the mouse preprogalanin gene. Gene, 182, 71-75.
Kohchi C, Tsutsui K. (2000). Avian galanin: cloning of complementary DNAs and characterization of transcripts in different tissues. Journal of Experimental zoology, 287, 183-190.
Lamanna C, Assisi L, Costagliola A, Vittoria A, Botte V, Cecio A. (1999). Galanin in the lizard oviduct: it’s distribution and relationships with estrogen, VIP and oviposition. Life Sci, 65, 91-101.
Lamanna C, Assisi L, Lucini C, Botte V. (1999). Galanin-containing-neurons, in the gastrointestinal tract of the lizard Padarcis s. sicula, as components of anally projecting nerve pathways. Neuroscience Letters, 268, 93-96.
Landry M, Roche D, Vila-Porcile E, Calas A. (2000). Effects of centrally administered galanin (1-16) on galanin expression in the rat hypothalamus. Peptides, 21, 1725-1733.
Li D, Tsutsui K, Muneoka Y, Minakata H, Nomoto K. (1996). An oviposition-inducing peptide: isolation, localization and function of avian galanin in the quail oviduct. Endocrinology, 137, 1618-1626.
Marks DL, Lent KL, Rossmanith WG, Clifton DK, Steiner RA. (1994). Activation-dependent regulation of galanin gene expression in gonadotropin-releasing hormone neurons in the female rat. Endocrinology, 134, 1991-1998.
McDonald MP, Gleason TC, Robinson JK, Crawley JN. (1998). Galanin inhibits performance on rodent memory tasks. Ann N Y Acad Sci, 863, 305-322.
Miller MA. (1998). Regulation of galanin in memory pathways. Ann N Y Acad Sci, 863, 323-341.
Miller MA, Kolb PE, Raskind MA. (1993). Testosterone regulates galanin gene expression in the bed nucleus of the stria terminalis. Brain Res, 611, 338-341.
Naor Z, Clayton RN, Catt KJ. (1980). Characterization of gonadotropin-releasing hormone receptors in cultured rat pituitary cells. Endocrinology, 107, 1144-1152.
Niewiarowski PH, Balk ML, Londraville RL. (2000). Phenotypic effects of leptin in an ectotherm: a new tool to study the evolution of life histories and endothermy? JEB, 203, 295-300.
Nilsson CL, Brinkmalm A, Minthon L, Blennow K, Ekman R. (2001). Processing of neuropeptide Y, galanin, and somatostatin in the cerebrospinal fluid of patients with Alzheimer’s disease and frontotemporal dementia. Peptides, 22, 2105-2112.
Ponchon M, Lause P, Maiter D. (2000). In vitro effects of oestradiol on galanin gene expression in rat anterior pituitary cells. J Neuroendocrinol,12, 559-564.
Romanelli F, Fillo S, Isidori A, Gaudino S, Conte D. (1998). Galanin stimulates steroidogenesis in rat leydig cells. Life Sci, 63, 255-263.
Rossmanith WG, Marks DL, Clifton DK, Steiner RA. (1994). Induction of galanin gene expression in gonadotropin-releasing hormone neurons with puberty in the rat. Endocrinology, 135, 1401-1408.
Selvais PL, Denef JF, Adam E, Maiter DM. (1995). Sex-steroid control of galanin in the rat hypothalamic-pituitary axis. J Neuroendocrinol, 7,401-407.
Tatemoto K, Rökaeus Å, Jörnvall H, McDonald TJ, Mutt V. (1983). Galanin: a novel biologically active pepetide from porcine intestine. FEBS Lett, 164, 124-128.
Todd JF, Small CJ, Akinsanya KO, Stanley SA, Smith DM, Bloom SR. (1998). Galanin is a paracrine inhibitor of gonadotroph function in the female rat. Endocrinology, 139, 4222-4229.
Wang Y, Colon JM. (1994). Purification and primary structure of galanin from the alligator stomach. Peptides,15, 603-606.
Wang Y, Lance VA, Nielsen PF, Conlon JM. (1999). Neuroendocrine peptides (insulin, pancreatic polypeptide, neuropeptide Y, galanin, somatostatin, substance P, and neuropeptide γ) from the desert tortoise, Gopherus agassizii. Peptides, 20, 713-722.
Weiss JM, Bonsall RW, Demetrikopoulos MK, Emery MS, West CH. (1998). Galanin: a significant role in depression? Ann N Y Acad Sci, 863, 364-382.
Wiesenfeld-Hallin Z, Xu XJ. (1998). Galanin in somatosensory function. Ann N Y Acad Sci, 863, 383-389.
指導教授 高永旭(Yun-Hsi Kao) 審核日期 2002-7-18
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