CAJ | 학술논문

γ氨基丁酸(GABA)是脊髓背角、前角内主要的抑制性神经递质.为了更好地观察脊髓背角内GABA能神经元的形态和功能,本研究使用了两种谷氨酸脱羧酶67绿色荧光蛋白(GAD67-GFP)基因敲入小鼠,并观察了敲入小鼠脊髓内的GFP表达状况.用免疫荧光组织化学双标记方法显示脊髓内所有的GFP阳性神经元基本上都呈GAD67和GABA阳性;GFP阳性神经元在脊髓背角的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,而在脊髓背角其它部位及前角则呈散在分布.脊髓内GFP阳性神经元的分布与GABA能神经元的分布一致.本文作者等还进一步在GAD67-GFP敲入小鼠中观察了GFP和神经元标志物神经元核蛋白(NeuN)的共存状况.脊髓背角内GFP阳性神经元分别占Ⅰ、Ⅱ和Ⅲ层的NeuN阳性神经元的31.5%、33.3%和44.7%,与以往的GABA免疫组化研究结果基本一致.本研究表明GAD67 GFP基因敲入小鼠脊髓内的GFP在GAD67启动子的调节下正确地表达于GABA能神经元,该基因敲入小鼠可用于脊髓GABA能神经元的形态学特征和生理学特性及其发育规律等方面的研究.
γ안기정산(GABA)시척수배각、전각내주요적억제성신경체질.위료경호지관찰척수배각내GABA능신경원적형태화공능,본연구사용료량충곡안산탈최매67록색형광단백(GAD67-GFP)기인고입소서,병관찰료고입소서척수내적GFP표체상황.용면역형광조직화학쌍표기방법현시척수내소유적GFP양성신경원기본상도정GAD67화GABA양성;GFP양성신경원재척수배각적Ⅰ～Ⅲ층최위밀집,배각심층내측부급중앙관주위정중등밀도분포,이재척수배각기타부위급전각칙정산재분포.척수내GFP양성신경원적분포여GABA능신경원적분포일치.본문작자등환진일보재GAD67-GFP고입소서중관찰료GFP화신경원표지물신경원핵단백(NeuN)적공존상황.척수배각내GFP양성신경원분별점Ⅰ、Ⅱ화Ⅲ층적NeuN양성신경원적31.5%、33.3%화44.7%,여이왕적GABA면역조화연구결과기본일치.본연구표명GAD67 GFP기인고입소서척수내적GFP재GAD67계동자적조절하정학지표체우GABA능신경원,해기인고입소서가용우척수GABA능신경원적형태학특정화생이학특성급기발육규률등방면적연구.
γ-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in both dorsal and ventral horn of the spinalcord. In order to understand the morphology and function of GABAergic neurons in the spinal cord more precisely, two strains ofglutamate decarboxylase 67-green fluorescence protein (GAD67-GFP) knock-in mice were used. The expression of GFP in thespinal cord of knock-in mice was observed. The results of the immunofluoroscent histochemical double-staining study showedthat all GFP-positive neurons in the spinal cord were basically immunopositive for GAD67 and GABA. GFP positive neuronswere concentrated in laninae Ⅰ～Ⅲ of the spinal dorsal horn, presented in moderate amounts in the medial part of the deep dor-sal born and in the area around the central canal, and more sparsely distributed in the remainder of the dorsal horn and in the ven-tral horn. The distribution pattern of GFP expression neurons in the spinal cord was in consistent with that of GABAergic neurons. GAD67-GFP knock in mice were further used to investigate the colocalization of GFP with a neuronal marker, neuronalnuclei protein (NeuN). The proportion of GFP-positive neurons among the NeuN-immunopositive neurons in the spinal cord wasabout 31.5%, 33.3% and 44.7% in laminae Ⅰ , Ⅱ and Ⅲ of the spinal dorsal horn, respectively, which was close to those ob-tained from GABA immunohistochemical studies. Our results indicated that the GFP expression in the spinal cord of the GAD67-GFP knock-in mouse was regulated properly in GAD67-positive, GABAergic neurons. The knock in technique would be a usefultool for studying the morphology and physiological properties anddevelopmental pattern of GABAergic neurons in the spinalcord.