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携带小鼠酪氨酸酶基因的青鳉(Oryzias latipes)橙色转基因变体皮肤中野生型色素沉着的表达与传递。

Expression and transmission of wild-type pigmentation in the skin of transgenic orange-colored variants of medaka (Oryzias latipes) bearing the gene for mouse tyrosinase.

作者信息

Matsumoto J, Akiyama T, Hirose E, Nakamura M, Yamamoto H, Takeuchi T

机构信息

Department of Biology, Keio University, Yokohama, Japan.

出版信息

Pigment Cell Res. 1992 Nov;5(5 Pt 2):322-7. doi: 10.1111/j.1600-0749.1992.tb00556.x.

Abstract

Transgenic fish carrying a reconstructed mouse tyrosinase gene, mg-Tyrs-J, were produced by microinjecting the gene into the oocyte nucleus of an orange-colored variant of medaka (Oryzias latipes). Of 64 oocytes microinjected and subsequently inseminated, 13 embryos developed normally beyond hatching and three of them exhibited brown skin pigmentation in the adult as was commonly observed in the wild type of this species. Light and electron microscopic examination disclosed a ubiquitous distribution of typical melanophores in the skin of these transgenic fish. Judging from their population density and distribution pattern, it was presumed that melanogenesis in these fish was elicited in amelanotic melanophores that resided in the skin of the orange-colored fish of this variant. Immunofluorescence with use of the anti-mouse tyrosinase antiserum lacking reactivity to medaka tyrosinase clearly disclosed that the gene introduced was expressed in the melanophores of transgenic fish. Crosses of female transgenic fish and males from an orange-colored variant yielded offspring exhibiting wild-type or orange-colored pigmentation in a ratio of 1:1, thus implying that mg-Tyrs-J integrated into the medaka genome behaves like a dominant gene. Little melanogenesis was observed in xanthophores, leucophores and iridophores in transgenic fish, suggesting possible specificity in recognition of teleostean cell types (i.e., melanophores) by the regulatory region of the mouse tyrosinase gene.

摘要

通过将重组的小鼠酪氨酸酶基因mg-Tyrs-J显微注射到青鳉(Oryzias latipes)橙色变种的卵母细胞核中,培育出了携带该基因的转基因鱼。在64个显微注射并随后受精的卵母细胞中,有13个胚胎正常发育至孵化后,其中3个成年后表现出棕色皮肤色素沉着,这在该物种的野生型中很常见。光镜和电镜检查显示,这些转基因鱼的皮肤中普遍分布着典型的黑素细胞。从它们的种群密度和分布模式判断,推测这些鱼的黑色素生成是由存在于该变种橙色鱼皮肤中的无黑色素黑素细胞引发的。使用对青鳉酪氨酸酶无反应性的抗小鼠酪氨酸酶抗血清进行免疫荧光检测,清楚地表明导入的基因在转基因鱼的黑素细胞中表达。雌性转基因鱼与橙色变种的雄性杂交,产生的后代中野生型或橙色色素沉着的比例为1:1,这意味着整合到青鳉基因组中的mg-Tyrs-J表现得像一个显性基因。在转基因鱼的黄色素细胞、白色素细胞和虹彩细胞中几乎没有观察到黑色素生成,这表明小鼠酪氨酸酶基因的调控区域对硬骨鱼类细胞类型(即黑素细胞)的识别可能具有特异性。

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