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大鼠睾丸中的乳酸脱氢酶活性:冻干切片荧光测定法与吩嗪硫酸甲酯组织化学定位法的比较

Lactate-dehydrogenase activity in the rat testis: a comparison between fluorometric assay of freeze-dried sections and histochemical localization with phenazine methosulphate.

作者信息

Heywood L, Blackshaw A

出版信息

J Histochem Cytochem. 1978 Nov;26(11):967-72. doi: 10.1177/26.11.722053.

Abstract

The activity of lactate dehydrogenase (LDH) in freeze-dried sections of rat testes was determined by using a fluorometric assay method and found to be 4.47 +/- 0.23 moles/Kg dry weight/hr (MKDH +/- S.E.M.) in whole sections, 3.31 +/- 0.16 in tubules and 12.0 +/- 1.9 in interstitial tissue. The activities and regional variation are similar to those measured in nervous tissue and are well correlated with the histochemical localization of LDH activity when phenazine methosulphate (PMS) is not used as an electron carrier. LDH and lipoamide dehydrogenase activity have the same histochemical distribution and there is no nonspecific staining with either method. The use of PMS results in reduced dependence on substrate and coenzyme and does not indicate higher interstitial activity but may provide an indication of developing lactate metabolism in maturing sperm. It is recomended that methods with and without PMS be used in studies of LDH activity in the testis.

摘要

采用荧光测定法测定大鼠睾丸冻干切片中乳酸脱氢酶(LDH)的活性,结果发现,整个切片中LDH的活性为4.47±0.23摩尔/千克干重/小时(MKDH±标准误),肾小管中为3.31±0.16,间质组织中为12.0±1.9。这些活性及区域差异与在神经组织中测得的结果相似,并且在不使用吩嗪硫酸甲酯(PMS)作为电子载体时,与LDH活性的组织化学定位密切相关。LDH和硫辛酰胺脱氢酶活性具有相同的组织化学分布,两种方法均无非特异性染色。使用PMS可降低对底物和辅酶的依赖性,并未显示间质活性更高,但可能提示成熟精子中乳酸代谢的发展。建议在睾丸LDH活性研究中同时使用有PMS和无PMS的方法。

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