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大鼠乳腺组织及分离细胞在妊娠和哺乳期的钙调蛋白含量。

Calmodulin content of rat mammary tissue and isolated cells during pregnancy and lactation.

作者信息

Riss T L, Bechtel P J, Baumrucker C R

出版信息

Biochem J. 1984 May 1;219(3):927-34. doi: 10.1042/bj2190927.

Abstract

The calmodulin content of heat-treated extracts of rat mammary tissue and isolated cells was measured by using stimulation of cyclic nucleotide phosphodiesterase (PDE) activity and radioimmunoassay (r.i.a.) procedures. The calmodulin content of mammary tissue increased 2.5-fold near the time of parturition, remained at the elevated level during lactation, then, after the onset of involution, decreased to values similar to those measured from mammary tissue of pregnant rats. When tissue from 15 animals in different stages of pregnancy, lactation and involution were compared, the r.i.a. gave 2.6-fold higher results than the PDE assay. To investigate further the increase in calmodulin content of mammary tissue, secretory and myoepithelial cells were enzymically dissociated from rat mammary tissue during different stages of pregnancy, lactation and involution. Protein, DNA, lactose, glucose-6-phosphate dehydrogenase and alkaline phosphatase were assayed to characterize the cell fractions. By using r.i.a., the calmodulin content per mg of protein in isolated secretory-cell fractions was high near parturition, then decreased and remained relatively constant during lactation. The amount of calmodulin expressed per mg of DNA in secretory cells did not show a marked change near parturition, suggesting a constant amount of calmodulin per cell. The calmodulin content of myoepithelial cells dissociated from mammary tissue and measured by using r.i.a. was 6-fold lower than in secretory cells and remained relatively constant during the course of lactation. The changing levels of calmodulin in rat mammary tissue during development are suggested to be related to proliferation and destruction of secretory epithelial cells, events that occur near parturition and involution respectively.

摘要

采用环核苷酸磷酸二酯酶(PDE)活性刺激法和放射免疫分析法(r.i.a.)测定了大鼠乳腺组织和分离细胞经热处理后的提取物中的钙调蛋白含量。乳腺组织中的钙调蛋白含量在分娩临近时增加了2.5倍,在哺乳期保持在升高水平,然后在退化开始后,降至与妊娠大鼠乳腺组织测得的值相似。当比较处于妊娠、哺乳和退化不同阶段的15只动物的组织时,放射免疫分析法得到的结果比磷酸二酯酶分析法高2.6倍。为了进一步研究乳腺组织中钙调蛋白含量的增加情况,在妊娠、哺乳和退化的不同阶段,从大鼠乳腺组织中酶解分离出分泌细胞和肌上皮细胞。测定了蛋白质、DNA、乳糖、葡萄糖-6-磷酸脱氢酶和碱性磷酸酶,以表征细胞组分。通过放射免疫分析法,分离的分泌细胞组分中每毫克蛋白质的钙调蛋白含量在分娩临近时较高,然后下降,并在哺乳期保持相对恒定。分泌细胞中每毫克DNA表达的钙调蛋白量在分娩临近时没有明显变化,表明每个细胞中的钙调蛋白量恒定。用放射免疫分析法测定的从乳腺组织中分离出的肌上皮细胞的钙调蛋白含量比分泌细胞低6倍,并且在哺乳期过程中保持相对恒定。大鼠乳腺组织在发育过程中钙调蛋白水平的变化被认为与分泌上皮细胞的增殖和破坏有关,这两个事件分别发生在分娩临近和退化时。

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