Analyses of direct action of light on fish chromatophores

光对鱼类色素细胞的直接作用分析

• 批准号: 11640687
• 负责人: OSHIMA Noriko
• 金额: $ 1.73万
• 依托单位: Toho Unversity
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11640687/
• 关键词: Erythrophores; Indophores; Light sensitivity; Visual pigment; Cone red; Cone green; mRNA; Cyclic AMP(cAMP); ティラピア; Gタンパク質; シグナル伝達; 光受容タンパク質; ロドプシン

Results obtained for 3 years (1999-2001) are summarized as follows :1) Erythrophores present in fins of Nile tilapia respond to light. In spectral regions of 400-400 nm and 550-600 nm, pigment aggregation occurs, while dispersion is accelerated at wavelengths between 470 and 530 nm. The degree of such responses depends on the light intensity. The same responses are observed in cultured erythrophores, suggesting the direct action of light on these cells.2) We examined the expression of mRNA of visual pigments in fins only where erythrophores are present. The PCR primers were designed corresponding to the sequences of tilapia cone opsins (GenBank). RT-PCR demonstrated the expression of mRNAs of two opsins, red opsin and green opsin. Using these cDNA fragments as probes, screening of tail fin cDAN library was performed, and total sequences of the opsins were determined. With in situ hybridization using cRNA of red opsin as probe, the distribution of red opsin mRNA was investigated and the probe was recognized in erythrophores. However, further study is required.3) Using pertussis toxin, cholera toxin and other chemicals, we found that pigment aggregation or dispersion in response to light are mediated through Gi or Gs, respectively, leading to a decrease or an increase in intracellular cAMP level, respectively. Although we examined the expression of Gt mRNA in tilapia fins, such expression was not detected.4) In iridophores of the neon tetra, the minimum effective intensity of light is about 3.4 lx. Light with a peak, wavelength of 550 nm is more effective in inducing the photoresponse (a shift in the spectral peak of reflected light to longer wavelengths) compared with light at 500 or 600 nm. Light at 400 nm is less effective.

本研究通过3年的研究，获得了如下结果：1）尼罗罗非鱼鳍中的红细胞对光有反应。在400-400 nm和550-600 nm的光谱区域中，发生颜料聚集，而分散在470和530 nm之间的波长处加速。这种反应的程度取决于光的强度。在培养的红细胞中观察到相同的反应，表明光对这些细胞的直接作用。2）我们只检测了存在红细胞的鳍中视色素mRNA的表达。根据罗非鱼视蛋白基因序列（GenBank）设计引物，进行PCR扩增。RT-PCR结果显示，红视蛋白和绿色视蛋白的mRNA在细胞内均有表达。以这些cDNA片段为探针，对尾鳍cDNA文库进行了筛选，并测定了视蛋白的全序列。以红色视蛋白cRNA为探针进行原位杂交，研究了红色视蛋白mRNA在红细胞中的分布，并对探针进行了识别。3）利用百日咳毒素、霍乱毒素和其他化学物质，我们发现光诱导的色素聚集或分散分别通过Gi或Gs介导，导致细胞内cAMP水平的降低或升高。虽然我们检测了Gt mRNA在罗非鱼鳍中的表达，但没有检测到这种表达。4）在虹鳟的虹膜细胞中，最小有效光强约为3.4 lx。与500或600 nm的光相比，峰值波长为550 nm的光在诱导光响应（反射光的光谱峰向更长波长的偏移）方面更有效。400 nm的光效率较低。

项目成果

Sugimoto, M.et al.: "Apoptosis in skin pigment cells of the medaka, Oryzias latipes(teleostei), during long-term chromatic adaptation : the role of sympathetic innervation"Cell Tissue Res.. 301. 205-216 (2000)

Sugimoto, M.等人：“青鳉、Oryzias latipes(teleostei)皮肤色素细胞的凋亡，在长期染色适应过程中：交感神经支配的作用”Cell Tissue Res. 301. 205-216 (2000)

Oshima, N. and Sugimoto, M.: "Chromatophore responses and color change in teleost fishes"In : J. Matsumoto and M. Mizoguchi (eds) Pigment Cells, Keio University Press,Tokyo. 161-176 (2001)

Oshima, N. 和 Sugimoto, M.：“硬骨鱼类的色素反应和颜色变化”，见：J. Matsumoto 和 M. Mizoguchi（编辑）色素细胞，庆应义塾大学出版社，东京。

Sugimoto,M.,Uchida,N.and Hatayama,M.: "Apoptosis in skin pigment cells of the medaka.Oryzias latipes (Teleostei), during long-term chromatic adaptation"Cell Tissue Research. 301. 205-216 (2000)

Sugimoto, M.、Uchida, N. 和 Hatayama, M.：“青鳉皮肤色素细胞的凋亡。Oryzias latipes (Teleostei)，在长期的色彩适应过程中”细胞组织研究。

Sugimoto, M. et al.: "Apoptosis in skin pigment cells of the medaka, Oryzias latipes (teleostei), during long-term chromatic adaptation : the role of sympathetic innervation."Cell Tissue Res. 301. 205-216 (2000)

Sugimoto, M. 等人：“青鳉、Oryzias latipes (teleostei) 皮肤色素细胞的凋亡，在长期染色适应过程中：交感神经支配的作用。”细胞组织研究。

Oshima, N.: "Direct reception of light by chromatophores of lower vertebrates"Pigment Cell Res.. 14. 312-319 (2001)

Oshima, N.：“低等脊椎动物色素细胞直接接收光”Pigment Cell Res.. 14. 312-319 (2001)