APPLICATION OF STEREOLOGICAL ANALYSIS OF CELL-VOLUME TO ISOLATED MYOCYTES IN CULTURE WITH AND WITHOUT ADRENERGIC-INNERVATION
APPLICATION OF STEREOLOGICAL ANALYSIS OF CELL-VOLUME TO ISOLATED MYOCYTES IN CULTURE WITH AND WITHOUT ADRENERGIC-INNERVATION
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DOI：
10.1002/ar.1092310209
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发表时间：
1991-10-01
影响因子：
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通讯作者：
MARVIN, WJ
中科院分区：
文献类型：
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作者：
ATKINS, DL;ROSENTHAL, JK;MARVIN, WJ
利用以前仅限于组织和器官细胞的立体技术，对肾上腺素能神经支配后培养心肌细胞的结构变化进行了三维分析。在培养96小时计算两组细胞的细胞体积：分离的肌细胞和肾上腺素能神经元支配的肌细胞。细胞核、细胞质和细胞的相对体积和绝对体积通过整个细胞的系统取样切片和点计数取样技术进行定量。同样测定了细胞质中线粒体、肌节和其他细胞成分的体积。数据分析采用方差分析和随机区组设计，以控制各培养间的差异。肾上腺素能神经支配使细胞体积（X +/- SEM）增加44% （3,344 +/- 196-mu-m3至4,816 +/- 400-mu-m3, P = 0.007）。线粒体绝对体积在神经支配后显著增加（521 +/- 42 ~ 744 +/- 54 ~ m3, P < 0.01）。绝对肌节体积无显著变化（750 +/- 92 mu-m3至642 +/- 1061 mu-m3， P = 0.14）。其他细胞成分，定义为除线粒体和肌节外的所有细胞质成分，随着神经的支配而显著增加（1,739 +/- 166-mu-m3至3,097 +/- 338-mu-m3, P = 0.02）。神经支配后，细胞核和细胞质的相对体积保持不变。然而，线粒体的相对体积下降了6%，肌节占据的细胞质百分比下降了44%，其他细胞成分占据的体积增加了22%。这些发现支持使用立体分析作为一种手段来量化培养的肌细胞的细胞体积。细胞大小的显著增加，以及细胞内成分的变化，伴随着体外肾上腺素能神经支配在发育中的心肌细胞。
A three-dimensional analysis to evaluate structural changes in cultured cardiac myocytes following adrenergic innervation was performed using stereological techniques formerly limited to cells in tissue and organs. Cell volumes were calculated for two groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons. Relative and absolute volumes of the nucleus, cytoplasm, and cell were quantified by systematically sampling sections throughout the cell and by point count sampling techniques. Volumetric estimates were similarly determined for the mitochondria, sarcomeres, and other cellular components in the cytoplasm. Data were analyzed with ANOVA and randomized block design to control for variation among the cultures. Adrenergic innervation produced a 44% increase in cell volume, X +/- SEM, (3,344 +/- 196-mu-m3 to 4,816 +/- 400-mu-m3, P = 0.007). The absolute volume of mitochondria significantly increased after innervation (521 +/- 42-mu-m3 to 744 +/- 54-mu-m3, P < 0.01). Absolute sarcomere volume did not change significantly (750 +/- 92-mu-m3 to 642 +/- 1061-mu-m3, P = 0.14). Other cellular components, defined as all cytoplasmic components except mitochondria and sarcomeres, significantly increased with innervation (1,739 +/- 166-mu-m3 to 3,097 +/- 338-mu-m3, P = 0.02). The relative volume of the nucleus and the cytoplasm in the cell remained unchanged following innervation. However, the relative volume of mitochondria decreased by 6%, the percent of the cytoplasm occupied by the sarcomeres decreased by 44%, and the volume occupied by the other cellular components increased by 22%. These findings support the use of stereological analysis as a means to quantify cell volumes of cultured myocytes. Significant increases in cell size, as well as changes in intracellular composition, accompany in vitro adrenergic innervation in the developing cardiac myocyte.