Substance P content in cultured neonatal rat vagal sensory neurons: the effect of nerve growth factor.
Substance P content in cultured neonatal rat vagal sensory neurons: the effect of nerve growth factor.
复制标题
培养的新生大鼠迷走神经感觉神经元中 P 物质的含量：神经生长因子的作用。
DOI：
10.1016/0006-8993(88)90056-x
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Wheeler,FB
中科院分区：
文献类型：
--
作者：
MacLean,DB;Lewis,SF;Wheeler,FB
为开始研究迷走感觉神经P物质（SP）合成和释放的调控因素，建立了新生大鼠结状神经节培养物。在从结状神经节的小片段获得的微外植体培养物中，SP以低量存在：3周后，每孔141 ± 36 pg，每孔10个神经节当量。为了提高神经元存活，使用中性蛋白酶酶促解离结状神经节。第5天的估计存活率为20- 30%，每个接种的神经节有800-1200个存活神经元，此后缓慢下降。10-20%的神经元有特异性SP免疫染色，大多数神经元直径较小（18-22 μm）。SP含量在5天内较低，然后在14天后逐渐上升至80-150 pg/接种的神经节。向培养基中加入神经生长因子（NGF，100 ng/ml）并不改变神经元存活。然而，SP含量在存在NGF的情况下加倍，或在其撤除后迅速下降至对照水平的一半：例如，在用NGF培养12天后1185 ± 176 pg/孔与撤除NGF的第8天-12，592 ± 118 pg/孔，平均值±S.D.，P < 0.01。生长抑素，目前在六分之一的SP量，是不变的神经生长因子。在随后的研究中，将神经元接种到先前分离的大鼠心房细胞上，存活率增加了50%，但没有改变每个存活神经元的SP含量。这些研究表明，SP存在于大鼠迷走神经感觉神经元的解离培养物中;数量和估计的净合成率与先前在体内的观察结果一致。研究还表明，在结状神经节神经元中，SP含量而不是神经元存活受NGF调节。这种模型可能被证明是有价值的SP和其他感觉神经肽在这类重要的内脏传入神经元的研究。
To begin to study the factors regulating the synthesis and release of substance P (SP) in the sensory vagus nerve, cultures of neonatal rat nodose ganglia were developed. In microexplant cultures, obtained from small fragments of nodose ganglia, SP was present in low amounts: after 3 weeks,141 ± 36pg per well, 10 ganglia equivalents per well. To enhance neuron survival, nodose ganglia were enzymatically dissociated using neutral protease. Estimated survival at 5 days was 20–30%, with 800–1200 surviving neurons per plated ganglion, and decreased slowly thereafter. Specific SP immunostaining was present in 10–20% of neurons, mostly of small diameter (18–22 μm). SP content was low for 5 days then rose progressively after 14 days to 80–150 pg per plated ganglion. The addition of nerve growth factor (NGF, 100 ng/ml) to the culture medium did not alter neuron survival. However, SP content was doubled in the presence of NGF, or fell rapidly to one-half control levels following its withdrawal: e.g. following 12 days in culture with NGF1185 ± 176pg/well vs NGF withdrawn day 8–12,592 ± 118pg/well, mean±S.D., P < 0.01. Somatostatin, present in one-sixth the amount of SP, was unaltered by NGF. In subsequent studies, plating of neurons onto previously dissociated rat atriacytes increased survival by 50% but did not alter SP content per surviving neurons. These studies demonstrate that SP is present in dissociated cultures of rat vagal sensory neurons; the quantities and estimated net synthesis rate correspond to previous observations in vivo. The studies also demonstrate that SP content but not neuron survival are regulated by NGF in nodose ganglion neurons. This model may prove valuable for the study of SP and other sensory neuropeptides in this important class of visceral afferent neurons.