Matsumoto S G, Gruener R P, Kreulen D L
Department of Physiology, University of Arizona College of Medicine, Tucson 85724.
Neuroscience. 1993 Dec;57(4):1147-57. doi: 10.1016/0306-4522(93)90056-l.
We report here the neurotransmitter characteristics of neurons cultured from the same ganglia of fetal and embryonic guinea-pigs. Both the celiac ganglion and the superior mesenteric ganglion were examined. In a previous paper we described the neurotransmitter properties of adult guinea-pig prevertebral sympathetic neurons grown in dissociated cell culture, including the expression by these cells of immunoreactivity for tyrosine hydroxylase, neuropeptide Y and somatostatin. Tyrosine hydroxylase immunoreactivity was ubiquitously expressed in all fetal embryonic cultures, as was the case for adult neurons. Fetal-derived celiac and superior mesenteric gangli neurons displayed neuropeptide Y and somatostatin immunoreactivity in the same percentage of neurons as in adult cultures but at markedly lower levels. Embryonic neurons also expressed somatostatin immunoreactivity in roughly the same proportion of neurons as in adult and fetal cultures; however, the expression of neuropeptide Y immunoreactivity in both celiac and superior mesenteric gangli cultures was significantly different. Specifically, neuropeptide Y immunoreactivity in embryonic celiac cultures was greatly reduced in both the number of positive-labeled neurons and the amount of immunoreactive product, while neuropeptide Y immunoreactivity in embryonic superior mesenteric gangli cultures was markedly increased compared to their adult and fetal counterparts. The expression of neuropeptide Y immunoreactivity in celiac neurons was found to be specifically elevated by culturing the neurons in medium conditioned by disassociated vascular cells, this treatment having no effect on tyrosine hydroxylase or somatostatin immunoreactivity. Heart cell-conditioned medium did not effect neuropeptide Y or somatostatin immunoreactivity, although it did result in a significant reduction of tyrosine hydroxylase immunoreactivity and an increase in 5-hydroxytryptamine immunoreactivity. We conclude that the expression of neuropeptide Y immunoreactivity develops independently in cultures of adult and near-term fetuses but that embryonic neurons require interactions with target cells to express this phenotype. Neuropeptide Y immunoreactivity can be induced in embryonic sympathetic neurons by a target-derived factor(s).
我们在此报告从胎儿和胚胎豚鼠相同神经节培养的神经元的神经递质特征。研究了腹腔神经节和肠系膜上神经节。在之前的一篇论文中,我们描述了在解离细胞培养中生长的成年豚鼠椎前交感神经元的神经递质特性,包括这些细胞对酪氨酸羟化酶、神经肽Y和生长抑素的免疫反应性表达。酪氨酸羟化酶免疫反应性在所有胎儿和胚胎培养物中普遍表达,成年神经元也是如此。来自胎儿的腹腔和肠系膜上神经节神经元显示神经肽Y和生长抑素免疫反应性的神经元百分比与成年培养物中相同,但水平明显较低。胚胎神经元也以与成年和胎儿培养物大致相同的神经元比例表达生长抑素免疫反应性;然而,腹腔和肠系膜上神经节培养物中神经肽Y免疫反应性的表达存在显著差异。具体而言,胚胎腹腔培养物中神经肽Y免疫反应性在阳性标记神经元数量和免疫反应产物量方面均大大降低,而胚胎肠系膜上神经节培养物中神经肽Y免疫反应性与其成年和胎儿对应物相比明显增加。发现通过在由解离的血管细胞条件化的培养基中培养神经元,腹腔神经元中神经肽Y免疫反应性的表达会特异性升高,这种处理对酪氨酸羟化酶或生长抑素免疫反应性没有影响。心脏细胞条件培养基对神经肽Y或生长抑素免疫反应性没有影响,尽管它确实导致酪氨酸羟化酶免疫反应性显著降低和5-羟色胺免疫反应性增加。我们得出结论,神经肽Y免疫反应性的表达在成年和近足月胎儿培养物中独立发展,但胚胎神经元需要与靶细胞相互作用才能表达这种表型。神经肽Y免疫反应性可由靶源性因子诱导胚胎交感神经元产生。