Stimulation of the proliferation and differentiation of mouse pink-eyed dilution epidermal melanocytes by excess tyrosine in serum-free primary culture
Stimulation of the proliferation and differentiation of mouse pink-eyed dilution epidermal melanocytes by excess tyrosine in serum-free primary culture
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DOI：
10.1002/jcp.10085
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发表时间：
2002-05-01
影响因子：
5.6
通讯作者：
Mizoguchi, M
中科院分区：
文献类型：
--
作者：
Hirobe, T;Wakamatsu, K;Mizoguchi, M
将源自粉红眼稀释（p）基因座处的野生型小鼠（黑色，C57 BL/10 jHir-P/P）及其同源突变小鼠（粉红眼稀释，C57 BL/10 jHir-p/p）的新生背部皮肤的表皮细胞悬浮液与从原代培养开始补充有额外的L-酪氨酸（Tyr）的无血清黑素细胞生长培养基一起培养。L-Tyr以剂量依赖性方式抑制P/P黑素细胞的增殖，而L-Tyr刺激p/p成黑素细胞和黑素细胞的增殖，而与剂量无关。另一方面，L-Tyr以剂量依赖性方式刺激（PIP）或诱导（p/p）表皮黑素细胞的分化。在P/P和p/p成黑素细胞和黑素细胞中，用2.0 MM L-Tyr培养14天，观察到真黑素标记物吡咯-2，3，5-三羧酸（PTCA）和褐黑素标记物氨基羟基苯丙氨酸（AHP）的含量略有增加。每个PIP黑素细胞的总黑素体（阶段I、II、III和IV）的平均数目不被L-Tyr处理改变，但是阶段IV黑素体在总黑素体中的比例增加。相反，在p/p成黑素细胞和黑素细胞中，L-Tyr显著增加了II、III和IV期黑素体的数量以及III期黑素体的比例。p/p黑素细胞培养液中PTCA和真黑素前体5，6-二羟基吲哚-2-羧酸（DHICA）的含量明显高于PIP黑素细胞。而p/p黑素细胞培养液中AHP和褐黑素前体5-S-半胱氨酰多巴（5-S-CD）的含量增加，其趋势与P/P黑素细胞相似。这些结果表明，在原代培养的p/p黑素细胞被过量的L-Tyr诱导合成真黑素，但难以在黑素体中积累。(C)2002 Wiley-Liss，Inc.
The epidermal cell suspensions of the neonatal dorsal skin derived from wild type mouse at the pink-eyed dilution (p) locus (black, C57BL/10jHir-P/P) and their congenic mutant mouse (pink-eyed dilution, C57BL/10jHir-p/p) were cultured with a serum-free melanocyte growth medium supplemented with additional L-tyrosine (Tyr) from initiation of the primary culture. L-Tyr inhibited the proliferation of P/Pmelanocytes in a dose-dependent manner, whereas L-Tyr stimulated the proliferation of p/p melanoblasts and melanocytes regardless of dose. On the other hand, L-Tyr stimulated (PIP) or induced (p/p) the differentiation of epidermal melanocytes in a dose-dependent manner. In both P/P and p/p melanoblasts and melanocytes cultured with 2.0 MM L-Tyr for 14 days, slight increases in contents of eumelanin marker, pyrrole-2,3,5-tricarboxylic acid (PTCA) and pheomelanin marker, aminohydroxyphenylalanine (AHP) were observed. The average number of total melanosomes (stages I, II, III, and IV) per PIP melanocyte was not changed by L-Tyr treatment, but the proportion of stage IV melanosomes in the total melanosomes was increased. On the contrary, in p/p melanoblasts and melanocytes L-Tyr increased dramatically the number of stage II, III, and IV melanosomes as well as the proportion of stage III melanosomes. Contents of PTCA and eumelanin precursor, 5,6-dihydroxyindole-2-carboxylic acid (DHICA) of cultured media in p/p melanocytes were much more greatly increased than in PIP melanocytes. However, contents of AHP and pheomelanin precursor, 5-S-cysteinyldopa (5-S-CD) of cultured media in p/p melanocytes were increased in a similar tendency to P/Pmelanocytes. These results suggest that p/p melanocytes in the primary culture are induced to synthesize eumelanin by excess L-Tyr, but difficult to accumulate them in melanosomes. (C) 2002 Wiley-Liss, Inc.