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ANALYSIS OF REGULATIVE FUNCTION OF MELANIN-RELATED GENE EXPRESSION BY BIOCHEMICALAND CELLULAR BIOLOGICAL METHODS

生物化学和细胞生物学方法分析黑色素相关基因表达的调控功能

基本信息

批准号：
10670812
负责人：
ITO Shosuke
金额：
$ 2.05万
依托单位：
FUJITA HEALTH UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670812/
关键词：
Mouse Agouti Black pink-eyed dilution Melanocyte Melanosome Tyrosine Tyrosinase ピンクアンドダイリューション純粋培養

项目摘要

The role of the agouti(A) gene in the development of epidermal melanocytes by comparing the activity of proliferation and differentiation of epidermal melanocytes from black (C57BL/10JHir) and agouti (C57BL/10JHir-A/A) mice in serum-free primary culture. There was no significant difference between agouti and black melanocytes in the proliferative activity as well as the reactivity to differentiation-stimulating factors. However, cultured agouti melanocytes possessed numerous stage "III" melanosomes than cultured black melanocytes under the electron microscopic observations. On the contrary, agouti melanocytes possessed a small number of stage "IV" melanosomes as compared to black melanocytes. The maturation of melanosomes form stage "III" to "IV" in agouti melanocytes is thought to be inhibited. To understand the mechanism of the action of the pink-dyed dilution (p) gene on the proliferation and differentiation of the epidermal melanocytes of pink-dyed dilution mice (C57BL/10Hir-p/p),L-tyrosine(L-try), the substrate of tyrosine, was supplemented to a serum-free metanocyte growth medium from the initiation of primary culture, and its effects were examined. In pink-eyed dilution melanocytes cultured with L-tyr, the numbers of melanosomes in all stage("I","II","III",and "IV")were dramatically increased, despite the limited numbers of stage "I","II",and "III" melanosomes in control culture. The content of pyrrole-2,3,5-tricarboxylic and (PTCA, degradation product of eumelanin)both in cultured cells and conditioned media was increased 2-fold. These results suggest that the pink-eyed dilution gene is involved in regulating the melanin synthesis as well as melanosome formation affecting the retention of L-try in melanosomes.

通过比较无血清原代培养黑鼠（C57 BL/10 JHir）和黑鼠（C57 BL/10 JHir-A/A）表皮黑素细胞的增殖和分化活性，探讨agglutinase（A）基因在表皮黑素细胞发育中的作用。黑素细胞和黑素细胞的增殖活性以及对分化刺激因子的反应性无显著差异。然而，培养的黑素细胞具有大量的“III”期黑素小体比培养的黑色黑素细胞在电镜下观察。相反，agglomerate黑素细胞具有少量的阶段“IV”黑素体相比，黑色黑素细胞。黑素体从“III”期到“IV”期在黑素细胞中的成熟被认为受到抑制。为了解pink-dilution（p）基因对pink-dilution小鼠（C57 BL/10 Hir-p/p）表皮黑素细胞增殖和分化的作用机制，从原代培养开始，在无血清的后成细胞生长培养基中添加酪氨酸底物L-酪氨酸（L-try），观察其作用。在用L-tyr培养的红眼稀释黑素细胞中，尽管在对照培养物中阶段“I”、“II”和“III”黑素体的数量有限，但所有阶段（“I”、“II”、“III”和“IV”）的黑素体的数量显著增加。培养细胞和条件培养基中吡咯-2，3，5-三羧酸和（PTCA，真黑素的降解产物）的含量均增加2倍。这些结果表明，粉眼稀释基因参与调节黑素合成以及黑素体的形成，影响L-try在黑素体中的保留。