摘要
目的 研究NMDA受体亚单位NR1在长期培养大鼠海马脑片各区中免疫组织化学反应活性变化规律。方法 将新生 6～ 9天大鼠海马和齿状回切成 4 0 0 μm厚脑片 (即海马脑片 ) ,分别体外培养 1、2、3、4、5周时再作 2 0 μm厚冰冻切片 ,行NR1免疫组织化学反应和图像分析。 结果 NR1在培养各周海马脑片各区锥体细胞和颗粒细胞胞体的免疫反应均呈阳性 ,胞膜深染 ,胞质淡染。培养 1周时 ,其反应强度为CA3区 >CA1区 >齿状回(DG) ;之后则为CA1区 >CA3区 >DG ;3周时各区反应均达高峰。结论 NR1在长期培养大鼠海马脑片各区的免疫活性有时空差异 。
Objective To investigate the changes in immunohistochcemical reactivity of NMDA receptor subunit 1 (NR1) in long-term hippocampal organotypic tissue cultures (HOTCs).?Methods The hippocampal slices of 400 μm thickness, prepared from 6-9 days old pups of SD rats, were cultured separately for 1, 2, 3, 4, 5 weeks in vitro (wiv). They were then cut by cryostat microtomy into 20 μm sections, in which the immunohistochemical reactions of NR1 were tested by ABC method and were evaluated by LEICA QWin image analysis system.?Results The immunohistochemical reaction of NR1 was positive in all pyramidal and granular cells of rat HOTC at every wiv, with the cell membrane heavily stained, while the cytoplasm lightly stained. The order of staining densities of positive cells was CA3>CA1>DG at 1 wiv, and then changed into CA1>CA3>DG afterwards. The expression of NR1 reached its peak level in all subfields of HOTCs at 3 wiv.?Conclusion The immunohistochemical reactivity of NR1 in long-term HOTCs varies in a time-space dependent manner. This implys that structural and functional alterations of NR1 may occur in long-term HOTCs.
出处
《徐州医学院学报》
CAS
2002年第6期471-473,共3页
Acta Academiae Medicinae Xuzhou
基金
江苏省自然科学基金项目 (BK99192 )
江苏省麻醉学重点实验室开放课题(K990 37)
江苏省重点学科徐州医学院生物化学与分子生物学研究中心开放课题 (990 0 1)
