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分离的鸡胚心脏细胞中的氨基酸摄取

Amino acid uptake in isolated chick embryo heart cells.

作者信息

Guidotti G G, Lüneburg B, Borghetti A F

出版信息

Biochem J. 1969 Aug;114(1):97-105. doi: 10.1042/bj1140097.

Abstract
  1. The preparation of cell suspensions by treatment of chick embryo hearts with collagenase at various stages of development is described. 2. Measurements of oxygen consumption, incorporation of labelled leucine into protein and accumulation of labelled alpha-aminoisobutyric acid against a concentration gradient indicated a long-lasting viability of the isolated heart cells in vitro; a satisfactory preservation of subcellular structures, including plasma membrane, was assessed by electron-microscopic examination. 3. The rate of alpha-aminoisobutyric acid accumulation by cardiac cells isolated from hearts at different stages of embryological development decreased with aging; insulin stimulated the intracellular accumulation of this amino acid analogue. 4. Insulin increased the uptake by isolated heart cells of several (14)C-labelled naturally occurring amino acids; however, the fraction of amino acid taken up by the cells that was recovered free intracellularly, and therefore the concentration ratio (between intracellular water and medium), was enhanced by the hormone only with glycine, proline, serine, threonine, histidine and methionine. When isolated heart cells were incubated in the presence of a mixture of labelled amino acids, the addition of insulin increased the disappearance of radioactivity from the medium. 5. The general pattern of amino acid transport (in the absence and in the presence of insulin) in isolated cardiac cells was similar to that found in intact hearts, suggesting that the biological preparation described in this paper might be useful for studies of cell permeability and insulin action.
摘要
  1. 描述了在不同发育阶段用胶原酶处理鸡胚心脏来制备细胞悬液的方法。2. 对氧气消耗、标记亮氨酸掺入蛋白质以及标记α-氨基异丁酸逆浓度梯度积累的测量表明,分离的心脏细胞在体外具有持久的活力;通过电子显微镜检查评估了包括质膜在内的亚细胞结构得到了良好的保存。3. 从胚胎发育不同阶段的心脏分离的心脏细胞积累α-氨基异丁酸的速率随衰老而降低;胰岛素刺激了这种氨基酸类似物在细胞内的积累。4. 胰岛素增加了分离的心脏细胞对几种(14)C标记的天然存在氨基酸的摄取;然而,只有甘氨酸、脯氨酸、丝氨酸、苏氨酸、组氨酸和蛋氨酸,激素才会提高细胞摄取的氨基酸中细胞内游离回收的部分,因此提高了(细胞内水与培养基之间的)浓度比。当在标记氨基酸混合物存在下孵育分离的心脏细胞时,添加胰岛素会增加培养基中放射性的消失。5. 分离的心脏细胞中氨基酸转运的总体模式(在有无胰岛素的情况下)与完整心脏中的相似,这表明本文所述的生物学制剂可能对细胞通透性和胰岛素作用的研究有用。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a075/1184801/b7a6afa70d0b/biochemj00696-0108-a.jpg

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