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为了观察结状神经节与心肌细胞联合培养中神经元的迁移和肽类神经递质的合成,我们建立了Wistar大鼠迷走神经结状神经节与心肌细胞联合培养的模型。用倒置相差显微镜观察了结状神经节与心肌细胞联合培养不同时间的活细胞生长情况。用Holmes还原银染色法观察了联合培养72 h和96 h的标本中神经元和心肌细胞的生长以及神经元的迁移。用免疫组织化学法观察了联合培养72 h和96 h的标本中肽类神经递质—P物质(SP)和降钙素基因相关肽(CGRP)的合成。结果表明,结状神经节与心肌细胞联合培养中,神经元在形态学上的发育成熟可能需要72 h就可以完成,而神经递质合成的发育成熟则需要96 h。神经元在形态学上的发育成熟并不能代表神经递质合成的发育已经完成。
In order to observe the migration of neurons and the synthesis of peptide neurotransmitters during the co-culture of nodose ganglion cells and cardiomyocytes, we established a co-culture model of vagal nodose ganglion cells and cardiomyocytes in Wistar rats. Inverted phase contrast microscope was used to observe the growth of living cells of nodose ganglion and cardiomyocytes co-cultured at different times. Holmes reduction silver staining was used to observe the growth and neuronal migration of neurons and cardiomyocytes in specimens cultured for 72 h and 96 h. Immunohistochemistry was used to observe the synthesis of peptide neurotransmitter-P substance (SP) and calcitonin gene-related peptide (CGRP) in the co-cultured samples for 72 h and 96 h. The results showed that in the co-culture of nodose ganglion cells and cardiomyocytes, morphological development of neurons may take 72 h to complete, while neurotransmitter synthesis maturation requires 96 h. The morphological maturation of neurons does not represent the completion of the development of neurotransmitter synthesis.