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仙台病毒通透化兔中性粒细胞中嗜天青颗粒和特异性颗粒胞吐作用的差异调控

Differential control of azurophilic and specific granule exocytosis in Sendai-virus-permeabilized rabbit neutrophils.

作者信息

Barrowman M M, Cockcroft S, Gomperts B D

机构信息

Department of Experimental Pathology, University College London.

出版信息

J Physiol. 1987 Feb;383:115-24. doi: 10.1113/jphysiol.1987.sp016399.

Abstract
  1. Rabbit neutrophils were permeabilized by treatment with Sendai virus. This was monitored by fluorescence measurement of the formation of the adduct of deoxyribonucleic acid (DNA) with ethidium bromide. 2. On addition of Ca2+, buffered (with EGTA) in the micromolar concentration range to the permeabilized cells, secretion of beta-glucuronidase (marker of azurophilic granules) and lysozyme (marker of specific granules) occurs. Lactate dehydrogenase (cytosol marker) is retained. Half-maximal secretion of beta-glucuronidase occurs at approximately pCa 6.3; lysozyme secretion occurs at approximately pCa 6.6. 3. Secretion is dependent on the provision of nucleoside triphosphates to the permeabilized cells. There is an absolute requirement for adenosine 5'-triphosphate (ATP) for the secretion of lysozyme, but beta-glucuronidase secretion can be partly supported by other nucleoside triphosphates in the order guanosine 5'-triphosphate (GTP) greater than uridine 5'-triphosphate (UTP) = xanthosine 5'-triphosphate (XTP) greater than cytidine 5'-triphosphate (CTP). 4. Secretion from both granules is complete within 10 min of adding Ca2+ to the permeabilized cells. There is a delay before commencement of beta-glucuronidase secretion of approximately half a minute; the secretion of lysozyme has no measurable delay.
摘要
  1. 用仙台病毒处理使兔中性粒细胞通透化。这通过荧光测量溴化乙锭与脱氧核糖核酸(DNA)加合物的形成来监测。2. 向通透化细胞中加入微摩尔浓度范围、用乙二醇双四乙酸(EGTA)缓冲的Ca2+时,β-葡萄糖醛酸酶(嗜天青颗粒标志物)和溶菌酶(特异性颗粒标志物)会分泌。乳酸脱氢酶(胞质溶胶标志物)则保留。β-葡萄糖醛酸酶分泌的半最大量出现在约pCa 6.3处;溶菌酶分泌出现在约pCa 6.6处。3. 分泌依赖于向通透化细胞提供核苷三磷酸。溶菌酶分泌绝对需要腺苷5'-三磷酸(ATP),但β-葡萄糖醛酸酶分泌可部分由其他核苷三磷酸支持,顺序为鸟苷5'-三磷酸(GTP)>尿苷5'-三磷酸(UTP)=黄苷5'-三磷酸(XTP)>胞苷5'-三磷酸(CTP)。4. 向通透化细胞中加入Ca2+后10分钟内,两种颗粒的分泌均完成。β-葡萄糖醛酸酶分泌开始前有大约半分钟的延迟;溶菌酶分泌没有可测量的延迟。

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