Solubilization of a membrane factor that stimulates levels of substance P and choline acetyltransferase in sympathetic neurons.
Solubilization of a membrane factor that stimulates levels of substance P and choline acetyltransferase in sympathetic neurons.
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溶解刺激交感神经元中 P 物质和胆碱乙酰转移酶水平的膜因子。
DOI：
10.1073/pnas.84.23.8726
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发表时间：
1987
影响因子：
11.1
通讯作者：
Kessler,JA
中科院分区：
文献类型：
--
作者：
Wong,V;Kessler,JA
发育中的神经元将产生哪些神经递质的选择受到神经元微环境的影响。在这项研究中，我们表明神经元与膜相关分子的接触促进了肽能和胆碱能特征的表达。用成年大鼠脊髓或交感神经节来源的质膜处理培养的新生大鼠交感神经元，可诱导肽递质P物质的表达，并增加胆碱能生物合成酶胆碱乙酰转移酶的水平。这种递质刺激活性可被洗涤剂辛基葡萄糖苷从脊髓膜上溶解，但不能被Triton X-100溶解。用4M氯化钠也能从脊髓膜上提取胆碱乙酰转移酶和P物质的刺激活性，表明该活性物质是膜相关的，而不是固有的结构膜分子。胰酶或热处理的提取物破坏了刺激递质的活性，表明该因子含有蛋白质。活性也被透明质酸酶处理破坏，这表明活性物质可能含有糖胺聚糖。4M氯化钠提取物的胆碱乙酰转移酶刺激活性在校正的Sephadex G-75柱上以单峰形式洗脱，保留时间略短于25 kDa的标准品。活性峰的NaDodSO4/聚丙烯酰胺凝胶电泳图显示在29 kDa处有一条优势带。因此，接触性刺激交感神经元中P物质和胆碱乙酰转移酶的活性是神经元暴露于29 kDa膜相关因子的结果。
The choice of which neurotransmitters will be produced by a developing neuron is influenced by the microenvironment of the neuron. In this study we show that neuronal contact with membrane-associated molecules promotes expression of peptidergic and cholinergic traits. Treatment of cultured neonatal rat sympathetic neurons with plasma membranes derived from adult rat spinal cord or sympathetic ganglia induced expression of the peptide transmitter substance P and increased levels of the cholinergic biosynthetic enzyme choline acetyltransferase. The transmitter-stimulating activity could be solubilized from spinal cord membranes by the detergent octyl glucoside but not by Triton X-100. The choline acetyltransferase- and substance P-stimulating activity also could be extracted from spinal cord membranes by 4 M sodium chloride, suggesting that the active material is membrane associated rather than an intrinsic structural membrane molecule. Trypsin or heat treatment of the extract destroyed the transmitter-stimulating activity, indicating that the factor contains a protein. Activity also was destroyed by hyaluronidase treatment, suggesting that the active material may contain a glycosaminoglycan. The choline acetyltransferase-stimulating activity in the 4 M NaCl extract was eluted in a single peak from a calibrated Sephadex G-75 column with a retention time slightly less than that of a 25-kDa standard. NaDodSO4/polyacrylamide gel electrophoresis of the active peak revealed a predominant band at 29 kDa. Thus, contact-mediated stimulation of substance P and choline acetyltransferase activity in sympathetic neurons results from neuronal exposure to a 29-kDa membrane-associated factor.