Isolation of functionally active acini from bovine mammary gland.
Isolation of functionally active acini from bovine mammary gland.
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DOI：
10.3168/jds.s0022-0302(79)83287-7
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发表时间：
1979-04
影响因子：
3.5
通讯作者：
C. Park;J. J. Smith-J.;M. Sasaki;W. Eigel;T. W. Keenan
中科院分区：
文献类型：
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作者：
C. Park;J. J. Smith-J.;M. Sasaki;W. Eigel;T. W. Keenan
研究了从泌乳牛乳腺中获得高产量完整腺泡的条件以及分离腺泡的某些结构和功能特征。三种胶原酶浓度（100，150和200毫克/100毫升）和孵育期（40，60和90分钟）的双因素实验表明，增加这两个因素显着增加净腺泡产量。获得的腺泡的最大量，基于脱氧核糖核酸的含量，是原始组织的10.3%。在形态学上，组分主要由腺泡或大细胞团组成，几乎所有细胞都不含台盼蓝。在完全营养培养基中培养的腺泡将放射性亮氨酸掺入蛋白质中。当腺泡在不添加氨基酸的培养基中孵育时，合成蛋白的比活性与孵育时间呈负相关。在脉冲标记放射性L-亮氨酸超过16分钟，真正的标记腺泡蛋白4分钟后发生。脉冲追逐标记的顺序动力学表现出独特的反应模式，在体外腺泡系统。放线菌酮抑制腺泡蛋白质合成，而3 '，5'-环磷酸腺苷强烈刺激腺泡蛋白质合成。
Conditions to obtain high yields of intact acini from lactating bovine mammary glands and certain structural and functional characteristics of isolated acini were investigated. A two-factor experiment with three collagenase concentrations (100, 150, and 200 mg/100 ml) and incubation periods (40, 60, and 90 min) demonstrated that increases in both factors significantly increased net acini yield. Largest amounts of acini obtained, based on content of deoxyribonucleic acid, were 10.3% of the original tissue. Morphologically, fractions consisted primarily of acini or large cell clumps, and nearly all cells excluded trypan blue. Acini cultured in complete nutrient medium incorporated radioactive leucine into proteins. When acini were incubated in medium without supplemental amino acids, specific activity of synthesized proteins was correlated negatively with incubation time. During pulse labeling with radioactive L-leucine over 16 min, true labeling of acinar proteins occurred after 4 min. Sequential kinetics of pulse-chase labeling demonstrated a response pattern unique to the in vitro acinar system. Acinar protein synthesis was inhibited by cycloheximide and strongly stimulated by by 3',5'-cyclic adenosine monophosphate.