Basal and stimulated release of substance P from dissociated cultures of vagal sensory neurons.
Basal and stimulated release of substance P from dissociated cultures of vagal sensory neurons.
复制标题
迷走感觉神经元分离培养物中 P 物质的基础释放和刺激释放。
DOI：
10.1016/0006-8993(90)90093-q
复制
发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Hayes,L
中科院分区：
文献类型：
--
作者：
MacLean,DB;Wheeler,F;Hayes,L
P物质是一种广泛分布的11个氨基酸的神经肽，存在于多达20%的迷走神经感觉细胞体和从它们发出的纤维中。为了研究调节SP释放的因素，从新生大鼠获得迷走感觉（结状或结状/颈静脉）神经节，并使用中性蛋白酶分离。在胶原基质上接种的神经元的存活率在2周时为10-20%，当神经元接种在先前解离的大鼠心房细胞上时为20-30%。P物质含量在培养的最初几天是低的，然后线性上升到0.1-0.2 pg/存活神经元。物质P被释放到4.5 mM钾介质中，以0.036%/min的稳定速率。在50 mM K+补充培养基中，20 min内的总释放增加了5-8倍，稳态释放增加了4-5倍，达到0.15%/min。感觉神经元特异性兴奋性神经毒素，辣椒素，诱发SP释放量与50 mM K+相似。净K+和辣椒素诱发，但不是基础释放完全抑制3.5 mM氯化钴。缓激肽，1-100 nM，刺激SP释放2-4倍以上的基础水平。毛喉素和佛波醇酯也增加SP释放1.5-3倍的基础金额。总之，P物质以与体内相似的量存在于培养的迷走神经感觉神经元中，并且响应于感觉特异性刺激而释放。这些文化应该允许探索的一些组织特异性因素调节神经递质释放的感觉迷走神经。
Substance P, the widely distributed 11 amino acid neuropeptide, is present in up to 20% of vagal sensory cell bodies and the fibers emanating from them. To study the factors regulating the release of SP, vagal sensory (nodose or nodose/jugular) ganglia were obtained from neonatal rats and dissociated using neutral protease. Survival of plated neurons on collagen substrate was 10–20% at 2 weeks and 20–30% when neurons were plated over previously dissociated rat atriacytes. Substance P content was low in cultures for the first several days, then rose linearly to 0.1–0.2 pg/surviving neuron. Substance P was released into a 4.5 mM potassium medium at a steady rate of 0.036%/min. In 50 mM K+supplemented medium, total release during 20 min increased 5–8-fold and steady-state release increased 4–5-fold to 0.15%/min. The sensory neuron specific excitatory neurotoxin, capsaicin, evoked SP release in similar amounts to 50 mM K+. Both net K+- and capsaicin-evoked, but not basal release were completely inhibited by 3.5 mM cobalt chloride. Bradykinin, 1–100 nM, stimulated SP release 2–4 times above basal levels. Forskolin and phorbol ester also increased SP release 1.5–3 times basal amounts. In summary, substance P is present in cultured vagal sensory neurons in amounts similar to in vivo and is released in response to sensory specific stimuli. These cultures should allow exploration of some of the tissue specific factors regulating neurotransmitter release in the sensory vagus nerve.