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目的:以PPE-GFP转基因小鼠为研究工具,观察绿色荧光蛋白(GFP)阳性的脑啡肽(ENK)能神经元与1型囊泡膜谷氨酸转运体(VGLUT1)在脊髓的分布及共存情况。方法:利用免疫组织化学和原位杂交双标染色的方法。结果:GFP标记的ENK能神经元主要位于脊髓背角,在I-ⅡI层最为密集,背角深层内侧部及中央管周围呈中等密度分布,散在分布于前角。VGLUT1 mRNA阳性细胞广泛分布在脊髓各层。GFP/VGLUT1双标细胞主要分布在脊髓背角,I-ⅡI层双标细胞占GFP阳性细胞的22.95±1.10%,占VGLUT1阳性细胞的27.91±2.42%;IV-VI层中21.49±4.99%GFP阳性细胞表达VGLUT1,10.35±2.81%VGLUT1阳性细胞表达GFP;前角双标细胞占VGLUT1阳性细胞的1.07±0.37%,占GFP阳性细胞的32.08±13.15%。结论:双标结果表明脊髓内部分ENK能神经元表达1型囊泡膜谷氨酸转运体,推测ENK能神经元可能通过调控谷氨酸的释放发挥感觉信息调控作用。
OBJECTIVE: To investigate the distribution of green fluorescent protein (GFP) -positive enkephalin (ENK) neurons and type 1 vesicular glutamate transporter (VGLUT1) in the spinal cord using PPE-GFP transgenic mice as a research tool. Coexistence. Methods: Immunohistochemistry and in situ hybridization double-stained method. RESULTS: The GFP-labeled ENK neurons were mainly located in the dorsal horn of spinal cord, and were most dense in I-III layer. The inner part of dorsal horn and the central canal were distributed in the anterior horn with moderate density. VGLUT1 mRNA positive cells are widely distributed in the spinal cord layers. The double-labeled cells of GFP / VGLUT1 mainly distributed in the dorsal horn of spinal cord. The double labeled cells in I-III layer accounted for 22.95 ± 1.10% of GFP-positive cells, accounting for 27.91 ± 2.42% of VGLUT1-positive cells and 21.49 ± 4.99% of GFP in IV- Positive cells expressed VGLUT1, while VGLUT1 positive cells expressed GFP at 10.35 ± 2.81%. Two anterior horn cells accounted for 1.07 ± 0.37% of VGLUT1 positive cells and 32.08 ± 13.15% of GFP positive cells. CONCLUSION: The double-labeled results show that ENK neurons in the spinal cord express a type 1 vesicular glutamate transporter, suggesting that ENK neurons may play a role in the regulation of sensory information by regulating the release of glutamate.