(3.238.96.184) 您好!臺灣時間:2021/05/08 03:04
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:李佩庭
研究生(外文):Pei-TingLi
論文名稱:探討 round-up 基因在果蠅骨骼肌發育的角色
論文名稱(外文):Characterizing the role of the round-up gene in Drosophila somatic muscle development
指導教授:劉雅心劉雅心引用關係
指導教授(外文):Ya-Hsin Liu
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生命科學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:46
中文關鍵詞:果蠅骨骼肌發育肌纖維母細胞融合
外文關鍵詞:Drosophila somatic muscle developmentmyoblast fusion
相關次數:
  • 被引用被引用:0
  • 點閱點閱:54
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
調控哺乳類動物和果蠅肌肉發育的基因網絡是高度保留的(conversed),因此可藉由遺傳學方式來探討果蠅骨骼肌發育過程以利於了解哺乳類動物肌肉的發育。本論文探討一個尚未被研究過的CG 基因。CG 基因在果蠅胚胎發育中期,除了會表現在骨骼肌前驅細胞中,也會表現在平滑肌前驅細胞中。為了要了解CG 在骨骼肌發育過程中所扮演的角色,利用 FRT-FLP 重組技術移除CG 並觀察CG 突變時的表現型態,結果發現,跟 wide type 相比,wide type 在胚胎發育後期會有肌纖維的形成,但
CG 突變時的表現型態卻沒有肌纖維的形成,反而呈現未融合的細胞狀態,因此將CG 基因命名為round - up 基因,推測round - up 基因可能跟調控骨骼肌的細胞融合有關。此外,也觀察round - up 基因過量表現時的肌肉發育形態,結果發現,在胚胎發育後期,肌纖維的數量有異常現象而且平滑肌也不能把中腸正常的收縮成四個腔室。為了探討CG 基因表現如何被調控,選殖round - up 基因的候選調控區域放置於報導基因綠螢光基因( GFP )前並建立帶有此片段的轉殖果蠅株,收集其胚胎並利用非螢光免疫染色觀察,結果發現,此候選調控區域表現形態跟內源性
CG 基因表現形態一致,表示此候選調控區域為round - up 基因的調控序列。
Somatic muscle development in Drosophila is analogous to mammals. To dissect Drosophila muscle development by genetics helps understand mammalian muscle development. I analyzed the role of an uncharacterized CG gene in mesoderm development during Drosophila embryogenesis. During Drosophila mid-embryogenesis, expression of CG was in the somatic muscle precursor cells and the visceral muscle precursor cells. In order to characterize the role of the CG gene in Drosophila somatic muscle development, I used FRT-FLP recombination to remove CG and examined phenotype of mutant CG embryos. Wild type embryos form muscle fibers at late embryogenesis, but I found that mutant embryos of the CG gene didn’t form muscle fibers. Instead, the founder cells remained unfused and appear in a rounded shape. I therefore named the gene round up. It implied that the gene round up may be involved in myoblast fusion in somatic muscle development. I then overexpressed round-up in mesoderm and found that embryos with round-up overexpression is defecting in the somatic muscle and abnormal constrictions in the visceral muscle during embryogenesis. To understand how CG gene expression is regulated, I cloned a candidate cis-regulatory region of round-up gene upstream a reporter gene GFP, and established transgenic fly carring this construct. I used non-fluorescence immunostaining to examine the expression of GFP which is driven by the candidate cis-regulatory region. I found that the expression pattern of the candidate genomic region was similar to round-up endogenous expression.
中文摘要……………………………………………………………………………1
英文延伸摘要………………………………………………………………………2
致謝…………………………………………………………………………………6
目錄…………………………………………………………………………………7
圖目錄………………………………………………………………………………10
縮寫表………………………………………………………………………………11
第一章、 前言……………………………………………………………………13
一. 果蠅中胚層形成…………………………………………………………13
二. 果蠅骨骼肌形成…………………………………………………………14
三. 果蠅平滑肌形成…………………………………………………………16
四. 實驗目的…………………………………………………………………17
第二章、 材料與方法……………………………………………………………18
一. 材料………………………………………………………………………18
(一) 實驗動物…………………………………………………………18
(二) 實驗藥品…………………………………………………………18
(三) 試劑組……………………………………………………………19
(四) 抗體………………………………………………………………19
(五) 限制酶……………………………………………………………20
二. 方法………………………………………………………………………20
(一) 建立經 FRT-FLP 的
D el
CG 果蠅株…………………………………20
1. 利用 FRT-FLP 來建構
D el CG 果蠅株………………………………20
2. 抽取果蠅基因組 DNA……………………………………………21
3. 聚合酶連鎖反應(Polymerase chain reaction)………………21

(二) 建立
UAS – cDNA of CG 轉殖果蠅株…………………………22
1. PCR 及酵素反應…………………………………………………22
2. 建構
UAS – cDNA of CG 質體…………………………………22
3. 質體 DNA 的萃取…………………………………………………23
(三) 建立含有 GFP 的候選調控區域的轉殖果蠅株……………………24
(四) 果蠅胚胎處理與免疫染色……………………………………………24
1. 果蠅胚胎收集………………………………………………………24
2. 果蠅胚胎固定………………………………………………………24
3. 螢光免疫染色………………………………………………………25
4. 非螢光免疫染色……………………………………………………25
5. 探針的製備…………………………………………………………26
6. 原位雜合反應………………………………………………………27
第三章、 結果………………………………………………………………………29
一. 鑑定Del CG 基因突變果蠅株…………………………………………………29
二. 觀察Del CG 基因表現型態……………………………………………………30
三. round - up 基因過量表現時的表現型態……………………………………30
四. 染色並觀察round - up 基因的候選調控區域表現形態……………………31
第四章、 討論…………………………………………………………………………32
一. UAS -cDNA of round-up是否會表現出有功能的Round-up…………32
二. round - up基因在骨骼肌發育過程可能參與的分子機制…………………32
三. round - up 基因會受哪些中胚層轉錄因子所調節……………………………33
第五章、 參考文獻……………………………………………………………………34
Abmayr SM, Pavlath GK (2012) Myoblast fusion: lessons from flies and mice. Development
139:641-656.
Azpiazu N, Lawrence PA, Vincent J-P, Frasch M (1996) Segmentation and specification of
the Drosophila mesoderm. Genes & Development 10:3183-3194.
Bate M, Rushton E (1993) Myogenesis and muscle patterning in Drosophila. Comptes
rendus de l'Academie des sciences Serie III, Sciences de la vie 316:1047-1061.
Baylies M, Arias AM, Bate M (1995) wingless is required for the formation of a subset of
muscle founder cells during Drosophila embryogenesis. Development
121:3829-3837.
Baylies MK, Bate M (1996) twist: a myogenic switch in Drosophila. Science 272:1481-1484.
Bodmer R (1993) The gene tinman is required for specification of the heart and visceral
muscles in Drosophila. Development 118:719-729.
Bour BA, O'Brien MA, Lockwood WL, Goldstein ES, Bodmer R, Taghert PH, Abmayr SM,
Nguyen HT (1995) Drosophila MEF2, a transcription factor that is essential for
myogenesis. Genes & development 9:730-741.
Boyle M, Bonini N, DiNardo S (1997) Expression and function of clift in the development
of somatic gonadal precursors within the Drosophila mesoderm. Development
124:971-982.
Castanon I, Von Stetina S, Kass J, Baylies MK (2001) Dimerization partners determine the
activity of the Twist bHLH protein during Drosophila mesoderm development.
Development 128:3145-3159.
Chen EH, Pryce BA, Tzeng JA, Gonzalez GA, Olson EN (2003) Control of myoblast fusion by
a guanine nucleotide exchange factor, loner, and its effector ARF6. Cell 114:751-762.
Deng H, Bell JB, Simmonds AJ (2010) Vestigial is required during late-stage muscle
differentiation in Drosophila melanogaster embryos. Molecular biology of the cell
21:3304-3316.
Elwell JA, Lovato TL, Adams MM, Baca EM, Lee T, Cripps RM (2015) The myogenic
repressor gene Holes in muscles is a direct transcriptional target of Twist and
Tinman in the Drosophila embryonic mesoderm. Developmental biology
400:266-276.
Furlong EE, Andersen EC, Null B, White KP, Scott MP (2001) Patterns of gene expression
during Drosophila mesoderm development. Science 293:1629-1633.
Kadam S, Ghosh S, Stathopoulos A (2012) Synchronous and symmetric migration of
Drosophila caudal visceral mesoderm cells requires dual input by two FGF ligands.
Development 139:699-708.
Kim JH, Jin P, Duan R, Chen EH (2015a) Mechanisms of myoblast fusion during muscle
development. Current opinion in genetics & development 32:162-170.
Kim JH, Ren Y, Ng WP, Li S, Son S, Kee Y-S, Zhang S, Zhang G, Fletcher DA, Robinson DN
(2015b) Mechanical tension drives cell membrane fusion. Developmental cell
32:561-573.
Lacroix C, Giovannini D, Combe A, Bargieri DY, Späth S, Panchal D, Tawk L, Thiberge S,
Carvalho TG, Barale J-C (2011) FLP/FRT-mediated conditional mutagenesis in
pre-erythrocytic stages of Plasmodium berghei. Nature protocols 6:1412-1428.
Lawrence PA, Bodmer R, Vincent J-P (1995) Segmental patterning of heart precursors in
Drosophila. Development 121:4303-4308.
Lilly B, Galewsky S, Firulli AB, Schulz RA, Olson EN (1994) D-MEF2: a MADS box
transcription factor expressed in differentiating mesoderm and muscle cell
lineages during Drosophila embryogenesis. Proceedings of the national academy
of sciences 91:5662-5666.
Lilly B, Zhao B, Ranganayakulu G, Paterson BM (1995) Requirement of MADS domain
transcription factor D-MEF2 for muscle formation in Drosophilia. Science 267:688.
Nguyen HT, Bodmer R, Abmayr SM, McDermott JC, Spoerel NA (1994) D-mef2: a
Drosophila mesoderm-specific MADS box-containing gene with a biphasic
expression profile during embryogenesis. Proceedings of the national academy of
sciences 91:7520-7524.
Park M, Wu X, Golden K, Axelrod JD, Bodmer R (1996) The Wingless Signaling Pathway Is
Directly Involved inDrosophilaHeart Development. Developmental biology
177:104-116.
Parks AL, Cook KR, Belvin M, Dompe NA, Fawcett R, Huppert K, Tan LR, Winter CG, Bogart
KP, Deal JE (2004) Systematic generation of high-resolution deletion coverage of
the Drosophila melanogaster genome. Nature genetics 36:288-292.
Ranganayakulu G, Schulz RA, Olson EN (1996) Wingless Signaling
InducesnautilusExpression in the Ventral Mesoderm of theDrosophilaEmbryo.
Developmental biology 176:143-148.
Richardson B, Beckett K, Baylies M (2008) Visualizing new dimensions in Drosophila
myoblast fusion. Bioessays 30:423-431.
Riechmann V, Irion U, Wilson R, Grosskortenhaus R, Leptin M (1997) Control of cell fates
and segmentation in the Drosophila mesoderm. Development 124:2915-2922.
Riechmann V, Rehorn K-P, Reuter R, Leptin M (1998) The genetic control of the distinction
between fat body and gonadal mesoderm in Drosophila. Development
125:713-723.
Rudolf A, Buttgereit D, Jacobs M, Wolfstetter G, Kesper D, Pütz M, Berger S,
Renkawitz-Pohl R, Holz A, Önel SF (2014) Distinct genetic programs guide
Drosophila circular and longitudinal visceral myoblast fusion. BMC cell biology
15:1.
Ruiz-Gomez M (1998) Muscle patterning and specification in Drosophila. INTERNATIONAL
JOURNAL OF DEVELOPMENTAL BIOLOGY 42:283-290.
Sandmann T, Jakobsen JS, Furlong EE (2006) ChIP-on-chip protocol for genome-wide
analysis of transcription factor binding in Drosophila melanogaster embryos.
Nature protocols 1:2839-2855.
Schnorrer F, Dickson BJ (2004) Muscle building: mechanisms of myotube guidance and
attachment site selection. Developmental cell 7:9-20.
Schulman VK, Dobi KC, Baylies MK (2015) Morphogenesis of the somatic musculature in
Drosophila melanogaster. Wiley Interdisciplinary Reviews: Developmental Biology
4:313-334.
Shirinian M (2009) Midgut and muscle development in Drosophila melanogaster.
Taylor MV (2000) Muscle development: molecules of myoblast fusion. Current Biology
10:R646-R648.
Taylor MV (2002) Muscle differentiation: how two cells become one. Current Biology
12:R224-R228.
Taylor MV (2003) Muscle differentiation: signalling cell fusion. Current Biology
13:R964-R966.
Taylor MV (2006) Comparison of muscle development in Drosophila and vertebrates. In:
Muscle development in Drosophila, pp 169-203: Springer.
Tixier V, Bataillé L, Jagla K (2010) Diversification of muscle types: recent insights from
Drosophila. Experimental cell research 316:3019-3027.
Wakabayashi-Ito N, Ip YT (2006) Mesoderm Formation in the Drosophila Embryo. In:
Muscle Development in Drosophila, pp 28-37: Springer.
Wu X, Golden K, Bodmer R (1995) Heart development in Drosophila requires the segment
polarity gene wingless. Developmental biology 169:619-628.
Zinzen RP, Girardot C, Gagneur J, Braun M, Furlong EE (2009) Combinatorial binding
predicts spatio-temporal cis-regulatory activity. Nature 462:65-70.
連結至畢業學校之論文網頁點我開啟連結
註: 此連結為研究生畢業學校所提供,不一定有電子全文可供下載,若連結有誤,請點選上方之〝勘誤回報〞功能,我們會盡快修正,謝謝!
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關論文
 
無相關期刊
 
無相關點閱論文
 
系統版面圖檔 系統版面圖檔