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用高效液相色谱法测定小鼠植入前胚胎中嘌呤补救和分解代谢的酶。

Enzymes of purine salvage and catabolism in the mouse preimplantation embryo measured by high performance liquid chromatography.

作者信息

Alexiou M, Leese H J

机构信息

Department of Biology, University of York, UK.

出版信息

J Reprod Fertil. 1994 May;101(1):151-8. doi: 10.1530/jrf.0.1010151.

Abstract

The activities of five enzymes involved in purine salvage and catabolism--hypoxanthine phosphoribosyl transferase (HPRT), adenine phosphoribosyl transferase (APRT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and guanase--were measured in mouse embryo extracts, from the one-cell to the blastocyst stage. Xanthine oxidase activity was not detected. The analyses were performed using high performance liquid chromatography and the enzymes showed different patterns of activity during development. Activities of HPRT, APRT and PNP were low before morula formation, and then increased until the blastocyst stage. ADA and guanase showed high activities after fertilization; guanase activity decreased sharply after the two-cell stage and ADA activity decreased sharply after the morula stage. Blastocyst formation was accompanied by a further decline in activity of both enzymes. The methods used may be suitable for measuring these enzymes in single human embryos, or in biopsies derived from them.

摘要

在小鼠胚胎提取物中,测定了从单细胞期到囊胚期参与嘌呤补救和分解代谢的五种酶——次黄嘌呤磷酸核糖转移酶(HPRT)、腺嘌呤磷酸核糖转移酶(APRT)、腺苷脱氨酶(ADA)、嘌呤核苷磷酸化酶(PNP)和鸟嘌呤酶的活性。未检测到黄嘌呤氧化酶活性。使用高效液相色谱法进行分析,这些酶在发育过程中表现出不同的活性模式。HPRT、APRT和PNP的活性在桑椹胚形成前较低,然后一直增加到囊胚期。ADA和鸟嘌呤酶在受精后活性较高;鸟嘌呤酶活性在二细胞期后急剧下降,ADA活性在桑椹胚期后急剧下降。囊胚形成伴随着这两种酶活性的进一步下降。所使用的方法可能适用于测量单个人类胚胎或源自它们的活检组织中的这些酶。

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