Pérez-García M T, Obeso A, López-López J R, Herreros B, González C
Departamento de Bioquímica y Biología Molecular y Fisiología, Facultad de Medicina, Universidad de Valladolid, Spain.
Am J Physiol. 1992 Dec;263(6 Pt 1):C1152-9. doi: 10.1152/ajpcell.1992.263.6.C1152.
Short-term cell cultures were obtained from enzymatically dissociated carotid bodies from adult rabbits, and morphological and functional characterization of the cultured chemoreceptor cells were carried out. Under phase contrast, freshly isolated type I cells are round, bright, and 10-14 microns in diameter and exhibit strong fluorescence when stained with the glyoxylic acid technique. The content of endogenous dopamine in the cultures increased from 80 pmol/10(5) cells 2 h after plating the cells to 200 pmol/10(5) cells on the 3rd day, and the rate of synthesis and storage of [3H]dopamine from the precursor [3H]tyrosine increased from 1.7 pmol.10(5) cells-1.h-1 in 1-day cultures to 4 pmol.10(5) cells-1.h-1 on the 3rd day; the later values represent 80-85% of the expected values for the intact carotid body. After labeling with [3H]tyrosine, cultured chemoreceptor cells release [3H]dopamine when challenged by hypoxia, high external K+, or the protonophore dinitrophenol, the pattern of response being similar to that of the intact carotid body. When studied by whole cell clamp recording, individual chemoreceptor cells exhibit a marked variability in the properties of some ionic currents; the data, however, do not support the existence of distinct subpopulations of type I cells.
从成年兔颈动脉体酶解分离获得短期细胞培养物，并对培养的化学感受细胞进行形态学和功能表征。在相差显微镜下，新鲜分离的I型细胞呈圆形、明亮，直径为10 - 14微米，用乙醛酸技术染色时呈现强荧光。培养物中内源性多巴胺的含量在细胞接种后2小时从80 pmol/10⁵细胞增加到第3天的200 pmol/10⁵细胞，并且从前体[³H]酪氨酸合成和储存[³H]多巴胺的速率从1天培养物中的1.7 pmol·10⁵细胞⁻¹·h⁻¹增加到第3天的4 pmol·10⁵细胞⁻¹·h⁻¹；后期的值代表完整颈动脉体预期值的80 - 85%。用[³H]酪氨酸标记后，培养的化学感受细胞在受到缺氧、高细胞外钾离子或质子载体二硝基苯酚刺激时释放[³H]多巴胺，反应模式与完整颈动脉体相似。当通过全细胞钳记录进行研究时，单个化学感受细胞在某些离子电流特性方面表现出明显的变异性；然而，数据不支持存在不同亚群的I型细胞。
