臺灣博碩士論文加值系統

由近來的研究中發現，一群bHLH-PAS蛋白在脊椎動物的胚胎發育過程中扮演著重要的角色。其中sim是bHLH-PAS蛋白群之一，並由研究証實與中樞神經系統的發育有密切關係。人類的sim2 gene位於人類第21條染色體上的唐氏症候群染色體區(Down syndrome critical region, DCR)，經研究証實過量表現hsim2所造成神經解剖學(neuroanatomic)及神經化學(neurochemical)異常發育的情況，與唐氏症病患的表現型相似。
先前，本實驗室成功的自斑馬魚(zebrafish)胚胎中選殖出一段sim基因，利用RNA全覆式原位雜交法觀察，結果顯示，SIM 會表現於間腦、頷弧和舌弧。為了進一步了解SIM 在斑馬魚胚胎發育過程中所扮演的角色，因此構築了三種不同的質體，在斑馬魚卵受精後的1-2個細胞時期，進行顯微注射，結果顯示，在注射pCMV-SIM2(150pg)及pCMV-AB(150pg)的斑馬魚胚胎，其顱底(neurocranium)及咽骨(mandibular arch , hyoid arch)的發育有異常的情形，接著更進一步利用專一性標示基因shh、msxB、dlx3、gsc進行RNA全覆式原位雜交法，發現其於斑馬魚胚胎中皆有異常的表現，因此本實驗証實SIM2基因在斑馬魚頭骨發育中扮演著重要的角色。

　　Recently , a novel bHLH-PAS protein family were revealed to have important roles in vertebrate development. Single-minded(sim) is a member of bHLH-PAS family that acts important role in central nervous system development. The human sim homolog , hsim2, was identified in the Down syndrome critical region of chromosome 21. It was proposed that the trisomy of the hsim2 gene induces the neuroanatomic and neurochemical abnormalities that often occur in DS.
Recently we have isolated a sim2 homolog gene from a zebrafish cDNA library (AF363019) .The expression pattern of zebrafish sim in the developing embryo was revealed by whole mount in situ hybridization. It shows that sim mRNA was detectable at diencephalons, mandibular arch and hyoid arch. To investigate the functions of sim in zebrafish development, we have microinjected a variety forms of sim DNA into fertilized embryo at 1-2 cell stages. Ectopic expression of sim2 gene or its C-terminal truncation form induced abnormal neurocranium and pharyngeal skeleton development and the expression pattern of midline regulator , shh , and downstream target of ET-1(endothelin-1) , including mexB, dlx3,gsc, were all disturbed by the ectopic sim2 expression , it suggests that sim2 gene plays important role in head skeleton development .

目錄
中文摘要……………………………………………………1
英文摘要…………………………………………………….2
壹、 緒論……………………………………………………4
貳、 實驗材料與方法………………………………………14
參、 結果……………………………………………………33
肆、 討論……………………………………………………39
伍、 參考文獻………………………………………………43
陸、 圖表……………………………………………………52

Akimenko, M.A., Ekker, M., Wegner, J., Lin, W. and Westerfield, M. (1994). Combinatorial expression of three zebrafish genes related to distall-less: part of a homeobox gene code for head. J. Neurosci. 10:3475-3486.
Ahlgren SC, Bronner-Fraser M.(1999) Inhibition of sonic hedgehog signaling in vivo results in craniofacial neural crest cell death. Curr Biol. 9(22):1304-14.
Akimenko MA, Johnson SL, Westerfield M, Ekker M.(1995) Differential induction of four msx homeobox genes during fin development and regeneration in zebrafish.Dev. 121(2):347-57.
Blagden CS, Currie PD, Ingham PW, Hughes SM.(1997) Notochord induction of zebrafish slow muscle mediated by Sonic hedgehog.Genes Dev. 11(17):2163-75.
Britto J, Tannahill D, Keynes R.(2002) A critical role for sonic hedgehog signaling in the early expansion of the developing brain. Nat Neurosci. 5(2):103-10.
Chandrasekhar A, Warren JT Jr, Takahashi K, Schauerte HE, van Eeden FJ, Haffter P, Kuwada JY.(1998) Role of sonic hedgehog in branchiomotor neuron induction in zebrafish. Mech Dev. 76(1-2):101-15.
Chandrasekhar A.Warren JT Jr.Takahashi K.Schauerte HE.van Eeden FJ.Haffter P. Chrast R, Scott HS, Madani R, Huber L, Wolfer DP, Prinz M, Aguzzi A, Lipp HP, Antonarakis SE.(2000) Mice trisomic for a bacterial artificial chromosome with the single-minded 2 gene (Sim2) show phenotypes similar to some of those present in the partial trisomy 16 mouse models of Down syndrome. Hum Mol Genet. 9(12):1853-64.
Chrast, R., Scott, H.S., Madani, R., Huber, L., Wolfer, D.P., Prinz, M., Aguzzi A., Lipp, H.P.(2000) Mice trisomic for a bacterial artificial chromosome with the sim2 gene show phenotypes similar to some of those present in the partial trisomy 16 mouse models of Down syndrome. Oxford University Press 9(12)1853-1864.
Crews ST, Fan CM.(1999) Remembrance of things PAS: regulation of development by bHLH-PAS proteins. Curr Opin Genet Dev9(5):580-7.
Dahmane N, Charron G, Lopes C, Yaspo ML, Maunoury C, Decorte L, Sinet PM, Bloch B, Delabar JM.(1995) Down syndrome-critical region contains a gene homologous to Drosophila sim expressed during rat and human central nervous system development. Proc Natl Acad Sci USA 26;92(20):9191-5.
Dambly-Chaudiere, C., Vervoort, M. 1998. The bHLH genes in neural development. Intl J Dev Biol 42: 269-273.
Ekker M, Akimenko MA, Allende ML, Smith R, Drouin G, Langille RM, Weinberg ES, Westerfield M.(1997) Relationships among msx gene structure and function in zebrafish and other vertebrates.Mol Biol Evol. 14(10):1008-22.
Ekker SC, Ungar AR, Greenstein P, von Kessler DP, Porter JA, Moon RT, Beachy PA.(1995) Patterning activities of vertebrate hedgehog proteins in the developing eye and brain. Curr Bio.5(8):944-55l
Ekker, M., Wegner, J., Akimenko, M.A. and Westerfield, M. (1992). Coordinate embryonic expression of three zebrafish engrailed genes. Development 116:1001-1010.
Ellies DL, Stock DW, Hatch G, Giroux G, Weiss KM, Ekker M.(1997) Relationship between the genomic organization and the overlapping embryonic expression patterns of the zebrafish dlx genes. Genomics45(3):580-90.
Ellies, D.L., Langille, R.M., Martin, C.C., Akimenko, M.A. and Ekker, M. (1996). Specific craniofacial cartilage dysmorphogenesis coincides with a loss of dlx gene expression in retinoic acid treated zebrafish embryos. Mech. Dev. 61:23-36.
Ellies, D.L., Stock, D.W., Hatch, G., Giroux, G., Weiss, K.M. and Ekker, M. (1997). Relationship between the genomic organization and the overlapping embryonic expression patterns of the zebrafish dlx genes. Genomics 45:580-590.
Ema M, Suzuki M, Morita M, Hirose K, Sogawa K, Matsuda Y, Gotoh O, Saijoh Y, Fujii H, Hamada H, Fujii-Kuriyama Y.(1996) cDNA cloning of a murine homologue of Drosophila single-minded, its mRNA expression in mouse development, and chromosome localization. Biochem Biophys Res Commun. 218(2):588-94.
Ema, M., Morita, M., Ikawa, S., Tanaka, M., Masuda, Y., Gotoh, O., Saijoh, Y., Ffujii, H., Hamada, H., Kikuchi, Y. and Yoshiaki, F.K. (1996). Two new members of the murine sim gene family are transcriptional repressors and show different expression patterns during mouse embryogenesis.Mol.Cell Biol. 16:5865-5875.
Epstein DJ, McMahon AP, Joyner AL.(1999) Regionalization of Sonic hedgehog transcription along the anteroposterior axis of the mouse central nervous system is regulated by Hnf3-dependent and -independent mechanisms.Dev. 126(2):281-92.
Epstein, D.J., Martinu, L., Michaud, J.L., Losos, K.L., Chen-Ming Fan and Alexandra,L.J.(2000) Members of the bHLH-PAS family regulate shh transcription in forebrain regions of the mouse CNS. Dev.(127)4701-4709.
Epstein, D.J., Martinu, L., Michaud, J.L., Losos, K.M., Fan, C.M. and Joyner, A.L. (2000). Members of the bHLH-PAS family regulate shh transcription in forebrain regions of the mouse CNS. Development 127:4701-4709.
Ericson J, Morton S, Kawakami A, Roelink H, Jessell TM.(1996) Two critical periods of Sonic Hedgehog signaling required for the specification of motor neuron identity.Cell 87(4):661-73.
Fan, C.M., Kuwana, E., Bulfone, A., Fletcher, C.F., Copeland, N.G., Jenkins, N.A., Crews, S., Martinez, S., Puelles, L., Rubenstein, J.L.R. and Marc, T.L. (1996). Expression patterns of two murine homologs of drosophila single-minded suggest possible roles in embryonic patterning and in the pathogenesis of Down syndrome. Mol. Cell Neurosci. 7:1-16.
Franks, R.G. and Crews, S.T. (1994). Transcriptional activation domains of the single-minded bHLH protein are required for CNS midline cell development. Mech. Dev. 45:269-277.
Gilbert,S.F. (1991). Early vertebrate development: neurulation and ectoderm. In Developmental Biology (3rd), pp. 165-197. Sinauer Press, Sunderland, Massachusetts.
Gilbert,S.F. (1991). Pattern formation. In Developmental biology (3rd), pp. 613-615. Sinauer Press, Sunderland, Massachusetts.
Gilbert-Barness E, Debich-Spicer D, Cohen MM Jr, Opitz JM.(2001) Evidence for the "midline" hypothesis in associated defects of laterality formation and multiple midline anomalies. Am J Med Gene. 101(4):382-7.
Goh KL, Yang JT, Hynes RO. (1997) Mesodermal defects and cranial neural crest apoptosis in alpha5 integrin-null embryos.Dev. 124(21):4309-19.
Hirakawa, T., Minegishi, T., Abe, K., Kishi, H., Ibuki, Y. and Miyamoto, K. (2000). Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the expression of luteinizing hormone receptors during cell differentiation in cultured granulose cells. A. Boil. Chem. Biop. 375(2):371-376.
Hjorth JT, Key B.(2001) Are pioneer axons guided by regulatory gene expression domains in the zebrafish forebrain? High-resolution analysis of the patterning of the zebrafish brain during axon tract formation. Dev Biol15;229(2):271-86.
Holder JL Jr, Butte NF, Zinn AR.(2000) Profound obesity associated with a balanced translocation that disrupts the SIM1 gene. Hum Mol Genet. 9(1):101-8.
Hu D.,Helms JA.(1999) The role of sonic hedgehog in normal and abnormal craniofacial morphogenesis.Development 126(21):4873-84
Izpisua-Belmonte JC, De Robertis EM, Storey KG, Stern CD.(1993) The homeobox gene goosecoid and the origin of organizer cells in the early chick blastoderm. Cell 74(4):645-59.
Klambt, C., Jacobs, J.R. and Goodman, C.S. (1991). The midline of the drosophila central nervous system: a model for the genetic analysis of cell fate, cell migration, and growth cone guidance. Cell 64:801-805.
Kondoh H, Uchikawa M, Yoda H, Takeda H, Furutani-Seiki M, Karlstrom RO.(2000) Zebrafish mutations in Gli-mediated hedgehog signaling lead to lens transdifferentiation from the adenohypophysis anlage. Mech Dev. 96(2):165-74.
Lewis, J.O. and Crews, S.T. (1994). Genetic analysis of the Drosophila single-minded gene reveals a central nervous system influence on muscle development. Mech. Dev. 48:81-91
Lindebro, M.C., Poellinger, L. and Whitelaw, M.L. (1995). Protein-protein interaction via PAS domains: role of the PAS domain in positive and negative regulation of the bHLH/PAS dioxin receptor-Arnt transcription factor complex. EMBO J. 14:3528-3539.
Litingtung Y, Chiang C.(2000) Control of Shh activity and signaling in the neural tube. Dev Dyn. 219(2):143-54.
Ma Y, Certel K, Gao Y, Niemitz E, Mosher J, Mukherjee A, Mutsuddi M, Huseinovic N, Crews ST, Johnson WA, Nambu JR.(2000) Functional interactions between Drosophila bHLH/PAS, Sox, and POU transcription factors regulate CNS midline expression of the slit gene. J Neurosci. 20(12):4596-605.
Marti E, Bovolenta P.(2002) Sonic hedgehog in CNS development: one signal, multiple outputs. 25(2):89-96.
Matise MP, Epstein DJ, Park HL, Platt KA, Joyner AL.(1998) Gli2 is required for induction of floor plate and adjacent cells, but not most ventral neurons in the mouse central nervous system.Development 125(15):2759-70.
Michaud, J.L., Derossi, C., May, N.R., Holdener, B.C. and Fan, C.M. (2000). ARNT2 acts as the dimerization partner of SIM1 for the development of the hypothalamus. Mech. Dev. 90:253-261.
Miller CT, Schilling TF, Lee K, Parker J, Kimmel CB.(2000) sucker encodes a zebrafish Endothelin-1 required for ventral pharyngeal arch development. Development 127(17):3815-28.
Moffett, P., Dayo, M., Reece, M., Mccormick, M.K. and Pelletier, J. (1996). Characterization of msim, a murine homologue of the Drosophila sim transcription factor. Genomics 35:144-155.
Moffett, p., Reece, M. and Pelletier, J. (1997). The murine sim2 gene product inhibits transcription by active repression and functional interference. Mol. Cell. Biol. 17:4933-4947.
Muenke M, Bone LJ, Mitchell HF, Hart I, Walton K, Hall-Johnson K, Ippel EF, Dietz-Band J, Kvaloy K, Fan CM, et al.(1995) Physical mapping of the holoprosencephaly critical region in 21q22.3, exclusion of SIM2 as a candidate gene for holoprosencephaly, and mapping of SIM2 to a region of chromosome 21 important for Down syndrome. Am J Hum Genet 57(5):1074-9.
Muller F, Albert S, Blader P, Fischer N, Hallonet M, Strahle U.(2000) Direct action of the nodal-related signal cyclops in induction of sonic hedgehog in the ventral midline of the CNS. Development 127(18):3889-97.
Muller F, Chang B, Albert S, Fischer N, Tora L, Strahle U.(1999) Intronic enhancers control expression of zebrafish sonic hedgehog in floor plate and notochord. Development 126(10):2103-16.
Muralidhar, M.G., Callahan, C.A. and Thomas, J.B. (1993). Single-minded regulation of genes in the embryonic midline of the Drosophila central nervous system. Mech. Dev. 41:129-138.
Nasrallah I, Golden JA.(2001) Brain, eye, and face defects as a result of ectopic localization of Sonic hedgehog protein in the developing rostral neural tube. Teratology64(2):107-13.
Odenthal, J. and Nusslein-Volhard, C. (1998). Fork head domain genes in zebrafish. Dev.Genes Evol. 208:245-258.
Orford J, Manglick P, Cass DT, Tam PP.(2001) Mechanisms for the development of esophageal atresia. J Pediatr Surg 36(7):985-94.
Piotrowski T, Schilling TF, Brand M, Jiang YJ, Heisenberg CP, Beuchle D, Grandel H, van Eeden FJ, Furutani-Seiki M, Granato M, Haffter P, Hammerschmidt M, Kane DA, Kelsh RN, Mullins MC, Odenthal J, Warga RM, Nusslein-Volhard C.(1996) Jaw and branchial arch mutants in zebrafish II: anterior arches and cartilage differentiation. Dev.123:345-56.
Richtsmeier JT, Zumwalt A, Carlson EJ, Epstein CJ, Reeves RH.(2002) Craniofacial phenotypes in segmentally trisomic mouse models for Down syndrome.Am J Med Genet107(4):317-24.
Rivera-Perez JA, Mallo M, Gendron-Maguire M, Gridley T, Behringer RR.(1995) Goosecoid is not an essential component of the mouse gastrula organizer but is required for craniofacial and rib development. Dev. 121(9):3005-12.
Sagerstrom, C.G., Grinbalt, Y. and Sive, H. (1996). Anteroposterior patterning in the zebrafish, Danio rerio: an explant assay reveals inductive and suppressive cell interactions. Development 122:1873-1883.
Schier AF, Neuhauss SC, Helde KA, Talbot WS, Driever W.(1997) The one-eyed pinhead gene functions in mesoderm and endoderm formation in zebrafish and interacts with no tail.Development 124(2):327-42.
Schilling TF, Piotrowski T, Grandel H, Brand M, Heisenberg CP, Jiang YJ, Beuchle D, Hammerschmidt M, Kane DA, Mullins MC, van Eeden FJ, Kelsh RN, Furutani-Seiki M, Granato M, Haffter P, Odenthal J, Warga RM, Trowe T, Nusslein-Volhard C.(1996) Jaw and branchial arch mutants in zebrafish I: branchial arches.Development 123:329-44.
Schilling TF.(1997) Genetic analysis of craniofacial development in the vertebrate embryo. Bioessays. 19(6):459-68.
Serluca, F.C., Fishman M.C.(2000) Pre-pattern in the pronephric kidney field of zebrafish.Development (128):2233-2241.
Stephen, T.C.(1998) Control of cell lineage-specific development and transcription by bHLH-PAS proteins. Gene Dev.Review.12:607-620.
Strahle U, Blader P, Ingham PW.(1996) Expression of axial and sonic hedgehog in wildtype and midline defective zebrafish embryos. Int J Dev Biol. 40(5):929-40.
Sun D, Baur S, Hay ED.(2000) Epithelial-mesenchymal transformation is the mechanism for fusion of the craniofacial primordia involved in morphogenesis of the chicken lip.Dev Bio. 228(2):337-49.
Swanson, H.I., Chan, W.K. and Bradfield, C.A. (1995). DNA binding specificities and pairing rules of the Ah receptor , ARNT , and SIM proteins. J. Biol.Chem. 270:26292-26302.
Wanner, R., Panteleyev, A., Henz, B.M. and Rosenbach, T. (1996). Retinoic acid affects the expression rate of the differentiation-related genes aryl hydrocarbon receptor, ARNT and keratin 4 in proliferative keratinocytes only. Biochim. et Biophy. Acta 1317:105-111.
Wen, H.J. (2000). “ Molecular cloning and expression of zebrafish single minded gene.” Master’s thesis, University of Taiwan Ocean, Keelung.
Woods SL, Whitelaw ML.(2002) Differential activities of murine single minded 1 (SIM1) and SIM2 on a hypoxic response element. Cross-talk between basic helix-loop-helix/per-Arnt-Sim homology transcription factors. J. B. Chem. 277(12):10236-43.
Yamaki A, Tochigi J, Kudoh J, Minoshima S, Shimizu N, Shimizu Y.(2001) Molecular mechanisms of human single-minded 2 (SIM2) gene expression: identification of a promoter site in the SIM2 genomic sequence. Gene 270(1-2):265-75.
Yamaki, A., Noda, S., Kudoh, J., Shindoh, N., Maeda, H., Minoshima, S., Kawasaki, K., Shimizu, N. and Shimiru, N. (1996). The mamalian single-minded (SIM) gene: mouse cDNA structure and diencephalic expression indicate a candidate gene for Down syndrome. Genomics 35:136-143.
Yamaki, A., Tochigi, J., Kudoh, J., Minoshima, S., Shimizu, N., Shimizu, Y.(2001) Molecular mechanisms of human sim2 gene expression: identification of a promoter site in the SIM2 genomic sequence. Gene 270:265-275.
Young DL, Schneider RA, Hu D, Helms JA.(2000) Genetic and teratogenic approaches to craniofacial development. Crit Rev Oral Biol Med. 11(3):304-17.
Zelzer, E., Wappner, P. and Shilo, B.Z. (1997). The PAS domain confers target gene specificity of Drosophila bHLH/PAS proteins. Genes Dev. 11:2079-2089.
Zerucha, T., Muller, J.P., Chartrand, N. and Ekker, M. (1997) Cross-interactions between two members of the dlx family of homeobox-containing genes during zebrafish development. Biochem. Cell Biol. 75:613-622.
Zhao, G.Q., Zhao, Q., Zhou, X., Mattei, M.G. and Crombrugghe, B. (1993) TFEC, a basic helix-loop-helix protein, forms heterodimers with TFE3 and inhibits TFE3-dependent transcription activation. Mol. Cell Biol. 13:4505-4512.