为观察坐骨神经慢性压迫性损伤(chronic constriction injury,CCI)后大鼠远位触液神经元(distal cerebrospinal fluid contacting neurons,dCSF-CNs)中drebrin的表达,探讨免疫荧光技术用于dCSF-CNs研究的可能性,将雄性Sprague-Dawley大鼠随机分为空白对照组、假手术组和CCI组,采用侧脑室注射霍乱毒素亚单位B(choleratoxin subunit B,CB)与辣根过氧化物酶(horseradish peroxidase,HRP)结合物(CB-HRP)示踪标记大鼠dCSF-CNs,观测三组大鼠行为学评分,并应用免疫荧光双标记和激光共聚焦显微镜技术比较各组dCSF-CNs中drebrin的表达。结果显示,三组中仅CCI组大鼠痛阈下降,三组大鼠dCSF-CNs均显示清晰,空白对照组和假手术组dCSF-CNs胞浆内无drebrin表达,CCI组dCSF-CNs胞浆内drebrin表达较多。结果表明,应用免疫荧光双标记观察dCSF-CNs,形态清晰,技术可靠。dCSF-CNs可能参与了神经病理性疼痛的信息传递。
To observe the expression of drebrin in the distal cerebrospinal fluid contacting neurons (dCSF-CNs) of rats with chronic constriction injury (CCI) of sciatic nerve by immunofluorescence technique, male Sprague-Dawley rats were randomly divided into three groups: control group, sham surgery group and CCI group. The behavior of rats was scored. After choleratoxin subunit B- conjugated horseradish peroxidase (CB-HRP, 3 μL) was injected into the lateral cerebroventricle to trace dCSF-CNs, the expression of drebrin was observed in the dCSF-CNs through immunofluorescence double staining and laser scanning confocal microscopy technique. The results showed that only the pain threshold of CCI group was decreased. The dCSF-CNs were clearly displayed in three groups. No drebrin expression was observed in the control and sham groups. In CCI group, drebrin was markedly expressed in intracytoplasm. It is suggested that the technique displaying dCSF-CNs with immunofluorescence is successful and the dCSF-CNs are possibly involved in the transmission of nociceptive information under the neuropathic pain state.