傷口癒合的能力對於生物體的生長很重要，目前已知JNK訊息傳導路徑會在傷口癒合中被活化，puckered (puc)是JNK訊息傳導路徑下游基因，會表現在傷口周圍。之前研究建立了包含有puc基因上游1.6 kb DNA片段的轉殖基因果蠅pucF43-pGP，發現此果蠅傷口附近的細胞質有pucF43-GFP的表現。經過transcription Element Search System (TESS)的預測，我們知道這段1.6 kb的序列中包含有AP-1轉錄因子結合位置，所以我們把puc基因上游0.7 kb (pucF3)和0.7 kb ~ 1.6 kb (pucF4)的DNA片段分別插入細胞核表現GFP的載體中，建立出兩種轉殖基因果蠅pucF3-pHS和pucF4-pHS，發現這兩種果蠅的GFP表現亮度、分布、時間和pucF43-pGP果蠅沒有明顯差異。我們利用pucF43-pGP果蠅來篩選哪些基因和傷口癒合相關。發現hep1會增加GFP的表現，並減緩傷口癒合速度。ed突變基因沒有明顯或是一致地改變GFP表現，所以我們推測ed不會影響傷口癒合中JNK訊息傳導。

The ability of wound healing for organisms is important. The downstream of JNK signaling pathway, puckered, was activated in wound healing process and expressed around the wound. We make a transgenic fly pucF43-pGP including the upstream 1.6 kb non-coding DNA sequences of puc and expressing pucF43-GFP in cytoplasm. This DNA fragment showed many AP-1 transcriptional factor binding sites by TESS. We inserted the upstream 0.7 kb (pucF3) and 0.7-1.6 kb (pucF4) non-coding DNA sequences of puc into pH-Stinger vector which expressing nuclear GFP to generate pucF3-pHS and pucF4-pHS transgenic flies. These two transgenic flies had no difference in GFP expressing pattern, GFP intensity and the speed of wound healing with pucF43-pGP fly significantly. We took pucF43-pGP transgenic fly to screen genes related to wound healing. hep1 increased the GFP intensity, and slowed down the speed of wound healing. ed mutant didn’t affected GFP expressing pattern or GFP intensity, so that we can not prove that ed related to wound healing.

參考資料

王天成 (2006) 果蠅傷口癒合過程中puckered表現之研究 國立清華大學 分子醫學研究所碩士論文
Bosch,M., Serras,F., Martin-Blanco,E., and Baguna,J. (2005). JNK signaling pathway required for wound healing in regenerating Drosophila wing imaginal discs. Dev. Biol. 280, 73-86.
Ciapponi,L., Jackson,D.B., Mlodzik,M., and Bohmann,D. (2001). Drosophila Fos mediates ERK and JNK signals via distinct phosphorylation sites. Genes Dev. 15, 1540-1553.
Escudero,L.M., Wei,S.Y., Chiu,W.H., Modolell,J., and Hsu,J.C. (2003). Echinoid synergizes with the Notch signaling pathway in Drosophila mesothorax bristle patterning. Development 130, 6305-6316.
Galko,M.J. and Krasnow,M.A. (2004). Cellular and genetic analysis of wound healing in Drosophila larvae. PLoS. Biol. 2, E239.
Glise,B., Bourbon,H., and Noselli,S. (1995). hemipterous encodes a novel Drosophila MAP kinase kinase, required for epithelial cell sheet movement. Cell 83, 451-461.
Harden,N. (2005). Cell biology. Of grainy heads and broken skins. Science 308, 364-365.
Islam,R., Wei,S.Y., Chiu,W.H., Hortsch,M., and Hsu,J.C. (2003). Neuroglian activates Echinoid to antagonize the Drosophila EGF receptor signaling pathway. Development 130, 2051-2059.
Jacinto,A., Woolner,S., and Martin,P. (2002). Dynamic analysis of dorsal closure in Drosophila: from genetics to cell biology. Dev. Cell 3, 9-19.
Kiehart,D.P., Galbraith,C.G., Edwards,K.A., Rickoll,W.L., and Montague,R.A. (2000). Multiple forces contribute to cell sheet morphogenesis for dorsal closure in Drosophila. J. Cell Biol. 149, 471-490.
Mace,K.A., Pearson,J.C., and McGinnis,W. (2005). An epidermal barrier wound repair pathway in Drosophila is mediated by grainy head. Science 308, 381-385.
Martin,P. (1997). Wound healing--aiming for perfect skin regeneration. Science 276, 75-81.
Martin-Blanco,E., Gampel,A., Ring,J., Virdee,K., Kirov,N., Tolkovsky,A.M., and Martinez-Arias,A. (1998). puckered encodes a phosphatase that mediates a feedback loop regulating JNK activity during dorsal closure in Drosophila. Genes Dev. 12, 557-570.
Mattila,J., Omelyanchuk,L., Kyttala,S., Turunen,H., and Nokkala,S. (2005). Role of Jun N-terminal Kinase (JNK) signaling in the wound healing and regeneration of a Drosophila melanogaster wing imaginal disc. Int. J. Dev. Biol. 49, 391-399.
Rawlins,E.L., Lovegrove,B., and Jarman,A.P. (2003). Echinoid facilitates Notch pathway signalling during Drosophila neurogenesis through functional interaction with Delta. Development 130, 6475-6484.
Redd,M.J., Cooper,L., Wood,W., Stramer,B., and Martin,P. (2004). Wound healing and inflammation: embryos reveal the way to perfect repair. Philos. Trans. R. Soc. Lond B Biol. Sci. 359, 777-784.
Riesgo-Escovar,J.R. and Hafen,E. (1997). Drosophila Jun kinase regulates expression of decapentaplegic via the ETS-domain protein Aop and the AP-1 transcription factor DJun during dorsal closure. Genes Dev. 11, 1717-1727.
Suzanne,M., Perrimon,N., and Noselli,S. (2001). The Drosophila JNK pathway controls the morphogenesis of the egg dorsal appendages and micropyle. Dev. Biol. 237, 282-294.
Ting,S.B., Caddy,J., Hislop,N., Wilanowski,T., Auden,A., Zhao,L.L., Ellis,S., Kaur,P., Uchida,Y., Holleran,W.M., Elias,P.M., Cunningham,J.M., and Jane,S.M. (2005). A homolog of Drosophila grainy head is essential for epidermal integrity in mice. Science 308, 411-413.
Wei,S.Y., Escudero,L.M., Yu,F., Chang,L.H., Chen,L.Y., Ho,Y.H., Lin,C.M., Chou,C.S., Chia,W., Modolell,J., and Hsu,J.C. (2005). Echinoid is a component of adherens junctions that cooperates with DE-Cadherin to mediate cell adhesion. Dev. Cell 8, 493-504.
Wood,W., Jacinto,A., Grose,R., Woolner,S., Gale,J., Wilson,C., and Martin,P. (2002). Wound healing recapitulates morphogenesis in Drosophila embryos. Nat. Cell Biol. 4, 907-912.