De Nollin S, Thoné F, Borgers M
J Histochem Cytochem. 1975 Oct;23(10):758-65. doi: 10.1177/23.10.172554.
The application of a new preparation method for demonstrating the activities of hydrolytic and oxidative enzymes in Candida albicans is reported. The problem of inadequate penetration of fixatives into yeast cells has been solved by sectioning solidified pellets of the cells in the presence of glutaraldehyde, a procedure that yields a fairly well preserved ultrastructure and sufficient enzyme activities. The subcellular distribution of most specific and nonspecific phosphatases and of peroxidases is at variance with that found in mammalian cells. The activities toward beta-glycerophosphate, p-nitrophenylphosphate, adenosine triphosphate, adenosine monophosphate, thiamine pyrophosphate and glucose 6-phosphate are almost exclusively confined to the central vacuolar apparatus. Oxidative and peroxidative activities are demonstrated only in mitochondria. Specific marker enzymes for endoplasmic reticulum, plasmalemma, Golgi apparatus and peroxisomes in C. albicans are not found. The possible function of the various subcellular organelles in relation to their enzymatic content is discussed.
本文报道了一种用于展示白色念珠菌水解酶和氧化酶活性的新制备方法的应用。通过在戊二醛存在下对固化的细胞团块进行切片，解决了固定剂难以充分渗透到酵母细胞中的问题，该方法能产生保存相当完好的超微结构和足够的酶活性。大多数特异性和非特异性磷酸酶以及过氧化物酶的亚细胞分布与哺乳动物细胞中的情况不同。对β-甘油磷酸、对硝基苯磷酸、三磷酸腺苷、一磷酸腺苷、硫胺焦磷酸和6-磷酸葡萄糖的活性几乎完全局限于中央液泡装置。氧化和过氧化活性仅在线粒体中得到证实。未发现白色念珠菌中内质网、质膜、高尔基体和过氧化物酶体的特异性标记酶。讨论了各种亚细胞器与其酶含量相关的可能功能。
