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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32476完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 胡孟君(Meng-Chun Hu) | |
| dc.contributor.author | Hui-Ting Lin | en |
| dc.contributor.author | 林慧婷 | zh_TW |
| dc.date.accessioned | 2021-06-13T03:51:40Z | - |
| dc.date.available | 2006-08-03 | |
| dc.date.copyright | 2006-08-03 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-25 | |
| dc.identifier.citation | Arukwe A. 2005. Modulation of brain steroidogenesis by affecting transcriptional changes of steroidogenic acute regulatory (StAR) protein and cholesterol side chain cleavage (P450scc) in juvenile Atlantic salmon (Salmo salar) is a novel aspect of nonylphenol toxicity. Environ Sci Technol 39:9791-9798.
Baulieu EE, Robel P. 1998. Dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEAS) as neuroactive neurosteroids. Proc Natl Acad Sci U S A 95:4089-4091. Beyenburg S, Watzka M, Blumcke I, Schramm J, Bidlingmaier F, Elger CE, Stoffel-Wagner B. 2000. Expression of mRNAs encoding for 17beta-hydroxisteroid dehydrogenase isozymes 1, 2, 3 and 4 in epileptic human hippocampus. Epilepsy Res 41:83-91. Cheng KC, Lee J, Khanna M, Qin KN. 1994. Distribution and ontogeny of 3 alpha-hydroxysteroid dehydrogenase in the rat brain. J Steroid Biochem Mol Biol 50:85-89. Chung BC, Hu MC, Lai CC, Lin CH. 1989. The 5'-region of the P450XIA1 (P450scc) gene contains a basal promoter and an adrenal-specific activating domain. Biochem Biophys Res Commun 160:276-281. Chung JH, Whiteley M, Felsenfeld G. 1993. A 5' element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila. Cell 74:505-514. Compagnone NA, Bulfone A, Rubenstein JL, Mellon SH. 1995. Expression of the steroidogenic enzyme P450scc in the central and peripheral nervous systems during rodent embryogenesis. Endocrinology 136:2689-2696. Compagnone NA, Mellon SH. 1998. Dehydroepiandrosterone: a potential signalling molecule for neocortical organization during development. Proc Natl Acad Sci U S A 95:4678-4683. Compagnone NA, Salido E, Shapiro LJ, Mellon SH. 1997. Expression of steroid sulfatase during embryogenesis. Endocrinology 138:4768-4773. Corpechot C, Robel P, Baulieu EE. 1981. Characterization and measurement of dehydroepiandrone sulfate in the rat brain. Proc. Natl. Acad. Sci. 78:4704-4707. Dupont E, Rheaume E, Simard J, Luu-The V, Labrie F, Pelletier G. 1991. Ontogenesis of 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase in the rat adrenal as revealed by immunocytochemistry and in situ hybridization. Endocrinology 129:2687-2692. Ebner MJ, Corol DI, Havlikova H, Honour JW, Fry JP. 2006. Identification of neuroactive steroids and their precursors and metabolites in adult male rat brain. Endocrinology 147:179-190. Emery DW, Yannaki E, Tubb J, Stamatoyannopoulos G. 2000. A chromatin insulator protects retrovirus vectors from chromosomal position effects. Proc Natl Acad Sci U S A 97:9150-9155. Foidart A, Harada N, Balthazart J. 1995. Aromatase-immunoreactive cells are present in mouse brain areas that are known to express high levels of aromatase activity. Cell Tissue Res 280:561-574. Fraile IG, McEwen BS, Pfaff DW. 1988. Comparative effects of progesterone and alphaxalone on aggressive, reproductive and locomotor behaviors. Pharmacol Biochem Behav 30:729-735. Furukawa A, Miyatake A, Ohnishi T, Ichikawa Y. 1998. Steroidogenic acute regulatory protein (StAR) transcripts constitutively expressed in the adult rat central nervous system: colocalization of StAR, cytochrome P-450SCC (CYP XIA1), and 3beta-hydroxysteroid dehydrogenase in the rat brain. J Neurochem 71:2231-2238. Greenstein BG, A. 2000. color atlas of neuroscience: neuroanatomy and neurophysiology. New York: Thieme. Griffin LD, Mellon SH. 1999. Selective serotonin reuptake inhibitors directly alter activity of neurosteroidogenic enzymes. Proc Natl Acad Sci U S A 96:13512-13517. Guarneri P, Guarneri R, Cascio C, Pavasant P, Piccoli F, Papadopoulos V. 1994. Neurosteroidogenesis in rat retinas. J Neurochem 63:86-96. Guennoun R, Fiddes RJ, Gouezou M, Lombes M, Baulieu EE. 1995. A key enzyme in the biosynthesis of neurosteroids, 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta-HSD), is expressed in rat brain. Brain Res Mol Brain Res 30:287-300. Guo IC, Tsai HM, Chung BC. 1994. Actions of two different cAMP-responsive sequences and an enhancer of the human CYP11A1 (P450scc) gene in adrenal Y1 and placental JEG-3 cells. J Biol Chem 269:6362-6369. Harada N, Yamada K. 1992. Ontogeny of aromatase messenger ribonucleic acid in mouse brain: fluorometrical quantitation by polymerase chain reaction. Endocrinology 131:2306-2312. He XY, Wegiel J, Yang SY. 2005. Intracellular oxidation of allopregnanolone by human brain type 10 17beta-hydroxysteroid dehydrogenase. Brain Res 1040:29-35. Heldt SA, Ressler KJ. 2006. Lesions of the habenula produce stress- and dopamine-dependent alterations in prepulse inhibition and locomotion. Brain Res 1073-1074:229-239. Hojo Y, Hattori TA, Enami T, Furukawa A, Suzuki K, Ishii HT, Mukai H, Morrison JH, Janssen WG, Kominami S, Harada N, Kimoto T, Kawato S. 2004. Adult male rat hippocampus synthesizes estradiol from pregnenolone by cytochromes P45017alpha and P450 aromatase localized in neurons. Proc Natl Acad Sci U S A 101:865-870. Hu MC, Chiang EF, Tong SK, Lai W, Hsu NC, Wang LC, Chung BC. 2001a. Regulation of steroidogenesis in transgenic mice and zebrafish. Mol Cell Endocrinol 171:9-14. Hu MC, Hsu HJ, Guo IC, Chung BC. 2004. Function of Cyp11a1 in animal models. Mol Cell Endocrinol 215:95-100. Hu MC, Hsu NC, Pai CI, Wang CK, Chung B. 2001b. Functions of the upstream and proximal steroidogenic factor 1 (SF-1)-binding sites in the CYP11A1 promoter in basal transcription and hormonal response. Mol Endocrinol 15:812-818. Hu ZY, Bourreau E, Jung-Testas I, Robel P, Baulieu EE. 1987. Neurosteroids: oligodendrocyte mitochondria convert cholesterol to pregnenolone. Proc Natl Acad Sci U S A 84:8215-8219. Hum DW, Staels B, Black SM, Miller WL. 1993. Basal transcriptional activity and cyclic adenosine 3',5'-monophosphate responsiveness of the human cytochrome P450scc promoter transfected into MA-10 Leydig cells. Endocrinology 132:546-552. Ibanez C, Guennoun R, Liere P, Eychenne B, Pianos A, El-Etr M, Baulieu EE, Schumacher M. 2003. Developmental expression of genes involved in neurosteroidogenesis: 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase in the rat brain. Endocrinology 144:2902-2911. Ivanova T, Beyer C. 2000. Ontogenetic expression and sex differences of aromatase and estrogen receptor-alpha/beta mRNA in the mouse hippocampus. Cell Tissue Res 300:231-237. Jung-Testas I, Hu ZY, Baulieu EE, Robel P. 1989. Neurosteroids: biosynthesis of pregnenolone and progesterone in primary cultures of rat glial cells. Endocrinology 125:2083-2091. Kaufman MH. 1995 The atlas of mouse development. San Diego Academic Press. Khanna M, Qin KN, Cheng KC. 1995. Distribution of 3 alpha-hydroxysteroid dehydrogenase in rat brain and molecular cloning of multiple cDNAs encoding structurally related proteins in humans. J Steroid Biochem Mol Biol 53:41-46. Kim HJ, Ha M, Park CH, Park SJ, Youn SM, Kang SS, Cho GJ, Choi WS. 2003. StAR and steroidogenic enzyme transcriptional regulation in the rat brain: effects of acute alcohol administration. Brain Res Mol Brain Res 115:39-49. Kimonides VG, Khatibi NH, Svendsen CN, Sofroniew MV, Herbert J. 1998. Dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEAS) protect hippocampal neurons against excitatory amino acid-induced neurotoxicity. Proc Natl Acad Sci U S A 95:1852-1857. Kimoto T, Tsurugizawa T, Ohta Y, Makino J, Tamura H, Hojo Y, Takata N, Kawato S. 2001. Neurosteroid synthesis by cytochrome p450-containing systems localized in the rat brain hippocampal neurons: N-methyl-D-aspartate and calcium-dependent synthesis. Endocrinology 142:3578-3589. King SL, Marks MJ, Grady SR, Caldarone BJ, Koren AO, Mukhin AG, Collins AC, Picciotto MR. 2003. Conditional expression in corticothalamic efferents reveals a developmental role for nicotinic acetylcholine receptors in modulation of passive avoidance behavior. J Neurosci 23:3837-3843. King SR, Manna PR, Ishii T, Syapin PJ, Ginsberg SD, Wilson K, Walsh LP, Parker KL, Stocco DM, Smith RG, Lamb DJ. 2002. An essential component in steroid synthesis, the steroidogenic acute regulatory protein, is expressed in discrete regions of the brain. J Neurosci 22:10613-10620. Kohchi C, Ukena K, Tsutsui K. 1998. Age- and region-specific expressions of the messenger RNAs encoding for steroidogenic enzymes p450scc, P450c17 and 3beta-HSD in the postnatal rat brain. Brain Res 801:233-238. Korneyev A, Pan BS, Polo A, Romeo E, Guidotti A, Costa E. 1993. Stimulation of brain pregnenolone synthesis by mitochondrial diazepam binding inhibitor receptor ligands in vivo. J Neurochem 61:1515-1524. Lauber ME, Lichtensteiger W. 1996. Ontogeny of 5 alpha-reductase (type 1) messenger ribonucleic acid expression in rat brain: early presence in germinal zones. Endocrinology 137:2718-2730. Le Goascogne C, Robel P, Gouezou M, Sananes N, Baulieu EE, Waterman M. 1987. Neurosteroids: cytochrome P-450scc in rat brain. Science 237:1212-1215. Majewska MD, Harrison NL, Schwartz RD, Barker JL, Paul SM. 1986. Steroid hormone metabolites are barbiturate-like modulators of the GABA receptor. Science 232:1004-1007. Mao X, Fujiwara Y, Orkin SH. 1999. Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice. Proc Natl Acad Sci U S A 96:5037-5042. Melcangi RC, Celotti F, Castano P, Martini L. 1993. Differential localization of the 5 alpha-reductase and the 3 alpha-hydroxysteroid dehydrogenase in neuronal and glial cultures. Endocrinology 132:1252-1259. Melcangi RC, Panzica G. 2003. Steroids and the nervous system. Introduction. Ann N Y Acad Sci 1007:1-5. Mellon SH, Deschepper CF. 1993. Neurosteroid biosynthesis: genes for adrenal steroidogenic enzymes are expressed in the brain. Brain Res 629:283-292. Mellon SH, Griffin LD, Compagnone NA. 2001. Biosynthesis and action of neurosteroids. Brain Res Brain Res Rev 37:3-12. Mellon SH, Miller WL, Bair SR, Moore CC, Vigne JL, Weiner RI. 1994. Steroidogenic adrenocortical cell lines produced by genetically targeted tumorigenesis in transgenic mice. Mol Endocrinol 8:97-108. Mensah-Nyagan AM, Feuilloley M, Do-Rego JL, Marcual A, Lange C, Tonon MC, Pelletier G, Vaudry H. 1996. Localization of 17beta-hydroxysteroid dehydrogenase and characterization of testosterone in the brain of the male frog. Proc Natl Acad Sci U S A 93:1423-1428. Mitev YA, Darwish M, Wolf SS, Holsboer F, Almeida OF, Patchev VK. 2003. Gender differences in the regulation of 3 alpha-hydroxysteroid dehydrogenase in rat brain and sensitivity to neurosteroid-mediated stress protection. Neuroscience 120:541-549. Modin C, Pedersen FS, Duch M. 2000. Lack of shielding of primer binding site silencer-mediated repression of an internal promoter in a retrovirus vector by the putative insulators scs, BEAD-1, and HS4. J Virol 74:11697-11707. Mukai H, Takata N, Ishii HT, Tanabe N, Hojo Y, Furukawa A, Kimoto T, Kawato S. 2006. Hippocampal synthesis of estrogens and androgens which are paracrine modulators of synaptic plasticity: synaptocrinology. Neuroscience 138:757-764. Naftolin F, Horvath TL, Jakab RL, Leranth C, Harada N, Balthazart J. 1996. Aromatase immunoreactivity in axon terminals of the vertebrate brain. An immunocytochemical study on quail, rat, monkey and human tissues. Neuroendocrinology 63:149-155. Nagy A. 2000. Cre recombinase: the universal reagent for genome tailoring. Genesis 26:99-109. Pelletier G, Luu-The V, Labrie F. 1994. Immunocytochemical localization of 5 alpha-reductase in rat brain. Mol Cell Neurosci 5:394-399. Plassart-Schiess E, Baulieu EE. 2001. Neurosteroids: recent findings. Brain Res Brain Res Rev 37:133-140. Romeo E, Cavallaro S, Korneyev A, Kozikowski AP, Ma D, Polo A, Costa E, Guidotti A. 1993. Stimulation of brain steroidogenesis by 2-aryl-indole-3-acetamide derivatives acting at the mitochondrial diazepam-binding inhibitor receptor complex. J Pharmacol Exp Ther 267:462-471. Sanne JL, Krueger KE. 1995. Expression of cytochrome P450 side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase in the rat central nervous system: a study by polymerase chain reaction and in situ hybridization. J Neurochem 65:528-536. Sharp PE, Turner-Williams S, Tuttle S. 2006. Movement-related correlates of single cell activity in the interpeduncular nucleus and habenula of the rat during a pellet-chasing task. Behav Brain Res 166:55-70. Sierra A. 2004. Neurosteroids: the StAR protein in the brain. J Neuroendocrinol 16:787-793. Soriano P. 1999. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21:70-71. Steckelbroeck S, Stoffel-Wagner B, Reichelt R, Schramm J, Bidlingmaier F, Siekmann L, Klingmuller D. 1999. Characterization of 17beta-hydroxysteroid dehydrogenase activity in brain tissue: testosterone formation in the human temporal lobe. J Neuroendocrinol 11:457-464. Steckelbroeck S, Watzka M, Reissinger A, Wegener-Toper P, Bidlingmaier F, Bliesener N, Hans VH, Clusmann H, Ludwig M, Siekmann L, Klingmuller D. 2003. Characterisation of estrogenic 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity in the human brain. J Steroid Biochem Mol Biol 86:79-92. Steckelbroeck S, Watzka M, Stoffel-Wagner B, Hans VH, Redel L, Clusmann H, Elger CE, Bidlingmaier F, Klingmuller D. 2001. Expression of the 17beta-hydroxysteroid dehydrogenase type 5 mRNA in the human brain. Mol Cell Endocrinol 171:165-168. Stoffel-Wagner B. 2003. Neurosteroid biosynthesis in the human brain and its clinical implications. Ann N Y Acad Sci 1007:64-78. Takayama K, Morohashi K, Honda S, Hara N, Omura T. 1994. Contribution of Ad4BP, a steroidogenic cell-specific transcription factor, to regulation of the human CYP11A and bovine CYP11B genes through their distal promoters. J Biochem (Tokyo) 116:193-203. Torres JM, Sanchez P, Ortega E. 2004. Quantitation of mRNA levels of steroid 5 alpha-reductase isoenzymes in the rat brain by 'one-step' RT-PCR and capillary electrophoresis. J Neurosci Methods 136:105-110. Tsuruo Y, Ishimura K, Fujita H, Osawa Y. 1994. Immunocytochemical localization of aromatase-containing neurons in the rat brain during pre- and postnatal development. Cell Tissue Res 278:29-39. Tsuruo Y, Miyamoto T, Yokoi H, Kitagawa K, Futaki S, Ishimura K. 1996. Immunohistochemical presence of 5 alpha-reductase rat type 1-containing cells in the rat brain. Brain Res 722:207-211. Ukena K, Usui M, Kohchi C, Tsutsui K. 1998. Cytochrome P450 side-chain cleavage enzyme in the cerebellar Purkinje neuron and its neonatal change in rats. Endocrinology 139:137-147. Uzunov DP, Cooper TB, Costa E, Guidotti A. 1996. Fluoxetine-elicited changes in brain neurosteroid content measured by negative ion mass fragmentography. Proc Natl Acad Sci U S A 93:12599-12604. Warner M, Gustafsson JA. 1995. Cytochrome P450 in the brain: neuroendocrine functions. Front Neuroendocrinol 16:224-236. Warner M, Tollet P, Stromstedt M, Carlstrom K, Gustafsson JA. 1989. Endocrine regulation of cytochrome P-450 in the rat brain and pituitary gland. J Endocrinol 122:341-349. Watanabe N, Inoue H, Fujii-Kuriyama Y. 1994. Regulatory mechanisms of cAMP-dependent and cell-specific expression of human steroidogenic cytochrome P450scc (CYP11A1) gene. Eur J Biochem 222:825-834. Wehrenberg U, Prange-Kiel J, Rune GM. 2001. Steroidogenic factor-1 expression in marmoset and rat hippocampus: co-localization with StAR and aromatase. J Neurochem 76:1879-1886. Wieland S, Lan NC, Mirasedeghi S, Gee KW. 1991. Anxiolytic activity of the progesterone metabolite 5 alpha-pregnan-3 alpha-o1-20-one. Brain Res 565:263-268. Yarim M, Kabakci N. 2004. Neurosteroidogenesis in oligodendrocytes and Purkinje neurones of cerebellar cortex of dogs. Anat Histol Embryol 33:151-154. Zhang P, Rodriguez H, Mellon SH. 1995. Transcriptional regulation of P450scc gene expression in neural and steroidogenic cells: implications for regulation of neurosteroidogenesis. Mol Endocrinol 9:1571-1582. 游祥明編著. 1999. 神經解剖學, first ed. 台北市: 藝軒圖書出版社. mouse brain library. http://www.mbl.org/atlas/atlas.php | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32476 | - |
| dc.description.abstract | 類固醇荷爾蒙主要於腎上腺、性腺與胎盤產生，近年已有研究發現腦部也能自行合成神經性類固醇，影響腦內的功能。在類固醇合成過程的第一步也是速率決定的步驟，是由P450膽固醇側鏈截切酶(P450scc)所催化。雖然前人利用RT-PCR技術測得此酵素的基因CYP11A1能在腦內表現，證明腦部具有新生類固醇的能力，但CYP11A1基因在腦內的表現量卻遠遠低於在腎上腺內的表現量，這使得P450scc在腦內的分佈區域和生理作用難以被確切描述。
本實驗室利用長度為4.4kb的人類CYP11A1啟動子控制Cre重組酶，產生了SCC-Cre的基因轉殖小鼠，目的是將來對類固醇生成組織內的基因做功能性的研究。我們發現Cre重組基因除了專一性表現在腎上腺與性腺，腦組織也有很好的Cre重組酶表現。在本篇研究中，我們分析Cre重組酶在腦內的活性與表現分布，結果顯示其主要分布在間腦區域的上丘腦與丘腦前端，以及中腦背部的上丘結構與頂蓋前區；在海馬迴結構中則有較弱的表現。我們進一步觀察到Cre重組基因在胚胎發育第10天的腦部開始有表現，並一直持續至出生前，其分布位置與成鼠的腦部一致。另外我們還觀察到在胚胎鼠的嗅覺上皮組織與視網膜內層有Cre表現。這些結果皆顯示轉殖的CYP11A1啟動子能驅動Cre重組酶在鼠腦內表現，且分布位置能反映目前已知的腦內內生性Cyp11a1基因的表現。鼠腦內生性Cyp11a1因表現量太低，難以被明確且直接的偵測，因此SCC-Cre小鼠提供了一個有價值的動物模式助於我們了解Cyp11a1在腦內可能的分布情形。另外， CYP11A1基因上游的4.4kb序列也是第一次被證實具有足以調控基因在腦部表現的元素。 | zh_TW |
| dc.description.abstract | Steroid hormones are mainly synthesized in adrenals, gonads and placenta. Recent studies have found that steroids can be de novo synthesized in brain, termed neurosteroid, to affect brain functions. The cholesterol side-chain cleavage enzyme (P450scc), encoded by CYP11A1 gene, catalyzes the first and rate-limiting step in steroid synthetic pathway. The expression of P450scc in the CNS has been detected by RT-PCR to demonstrate the steroidogenic capacity in the brain. However, the levels of CYP11A1 expression are much lower than that in adrenal glands. It is therefore difficult to identify the exact expressional pattern and also brings challenges to elucidate the physiological function of P450scc in brain.
We generated a SCC-Cre transgenic mouse, in which Cre recombinase expression is under the control of the human CYP11A1 promoter. It has been shown that Cre recombinanse was specifically expressd in the adrenals and gonads. In addition, we found that the brain tissues also express Cre protein in the SCC-Cre mice. By RT-PCR analysis, the expression of Cre was identified in all regions of brain except cerebellum. High levels of Cre expression was detected in the epithalamus and anterior thalamus of the diencephalons, and superior colliculus and pretectum of the midbrain. Weak expression was also found in the hippocampal formation. The Cre recombinase activity was first present in the embryonic brain at 10.5 days postcoitum in a pattern similar to that of adult brain. The Cre expression was further observed in the olfactory epithelium and the inner layer of retina. In conclusion, the 4.4 kb of 5’flanking region of the human CYP11A1 genes contains efficient regulatory elements to drive transgene expression in the mouse brain. Our SCC-Cre transgenic mice provide a valuable animal model to understand the possible expressional pattern of CYP11A1 in the brain. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T03:51:40Z (GMT). No. of bitstreams: 1 ntu-95-R93441011-1.pdf: 2386721 bytes, checksum: df26ef025fb9dc89fd75a7d653fe0f55 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 表次 ……………………………………………………………………….. IV
圖次 ………………………………………………………………………... IV 腦解剖構造中英文名稱對照表 …………………………………………... V 中文摘要 …………………………………………………………………… VIII 英文摘要 …………………………………………………………………... IX 第一章 前言 ………………………………………………………………. 1 一、類固醇的生成途徑 …………………………………………….. 1 二、神經性類固醇的生成及其功能 ………………………………… 2 1、 神經性類固醇的生成途徑 ………………………………... 2 2、 神經性類固醇的功能 …………………………………….. 3 三、腦內細胞色素P450膽固醇側鏈截切酶 (Cyp11a1)的研究 …... 4 1、 Cyp11a1在腦內的表現量 ……………………………….. 4 2、 Cyp11a1在腦內的分布 …………………………………… 5 3、 Cyp11a1在腦內的活性與生理意義 ……………………... 6 4、 研究腦內Cyp11a1功能與基因調控的困難 …………….. 7 四、Cre/loxP系統於組織專一性的基因剔除之應用 ………………. 8 1、 建立方式與基因功能研究的應用 ………………………. 8 2、 Cre重組酶表現位置與活性的檢測 …………………….. 9 五、人類CYP11A1啟動子於生物體內的研究 ……………………. 9 1、 細胞株實驗模式 ………………………………………… 9 2、 基因轉殖動物模式 ………………………………………. 10 3、 本實驗室建立的SCC-Cre動物模式 …………………… 11 六、研究動機與目的 ………………………………………………… 12 第二章 材料與方法 …………………………………………………….. 12 一、轉殖基因的建構 ………………………………………………… 12 二、SCC-Cre基因轉殖小鼠的產生 ………………………………… 12 三、SCC-Cre/R26R小鼠之交配與鑑定 ……………………………. 12 四、小鼠基因型檢測 (Genotyping) ………………………………… 13 五、小鼠不同胚胎時期的判定 ……………………………………... 13 六、灌流 ……………………………………………………………... 14 七、取腦 ……………………………………………………………. 14 八、冷凍組織切片 …………………………………………………... 15 九、X-gal酵素活性染色 …………………………………………… 15 十、反轉錄-聚合酵素連鎖反應 (RT-PCR) ………………………… 15 1. 組織準備 ……………………………………………………. 15 2. 抽取RNA …………………………………………………… 16 3. DNase處理 …………………………………………………... 16 4. 反轉錄作用 ………………………………………………….. 17 5. 聚合酵素鏈鎖反應 (PCR) …………………………………. 17 第三章 結果 ………………………………………………………………. 19 一、Cre重組酶在成鼠腦內的表現分布 …………………………… 19 二、Cre重組酶在胚胎鼠腦內之表現 ……………………………… 22 三、內生性Cyp11a1與轉殖基因在小鼠腦內表現之比較 ………. 24 第四章 討論 ……………………………………………………………… 26 一、人類CYP11A1基因上游的4.4kb片段足以調控轉殖基因在腦 組織表現 ...................................................................................... 26 二、以Cre重組酶在SCC-Cre小鼠的表現預測內生性Cyp11a1在 腦內表現布 ................................................................................... 28 三、人類CYP11A1基因5’端4.4kb可以調控胚胎時期腦內內源基 因的表現 ……………………………………………………….. 29 四、由Cre重組酶在腦內表現之特徵探討Cyp11a1可能的神經功 能 ………………………………………………………………… 30 參考文獻 ……………………………………………………………… 33 表 ……………………………………………………………………… 43 圖 ……………………………………………………………………… 44 | |
| dc.language.iso | zh-TW | |
| dc.subject | 鼠腦 | zh_TW |
| dc.subject | Cre重組酶 | zh_TW |
| dc.subject | CYP11A1啟動子 | zh_TW |
| dc.subject | CYP11A1 promoter | en |
| dc.subject | mouse brain | en |
| dc.subject | Cre recombinase | en |
| dc.title | 長度4.4kb人類CYP11A1啟動子能驅動Cre重組酶在鼠腦內表現 | zh_TW |
| dc.title | Human CYP11A1 4.4kb promoter drives Cre recombinase expression in the mouse brain | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳儀莊(Yi-juang Chern),郭應誠(Ing-Cherng Guo),湯志永(Chih-Yung Tang) | |
| dc.subject.keyword | CYP11A1啟動子,Cre重組酶,鼠腦, | zh_TW |
| dc.subject.keyword | CYP11A1 promoter,Cre recombinase,mouse brain, | en |
| dc.relation.page | 60 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-26 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 生理學研究所 | zh_TW |
| 顯示於系所單位： | 生理學科所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-95-1.pdf 未授權公開取用 | 2.33 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。