豚鼠视网膜组织M4受体的表达

目的 检测M4受体在豚鼠视网膜组织的表达及分布.方法 4周龄三色豚鼠21只,其中7只动物眼球制备石蜡切片,免疫组织化学染色法检测后极部视网膜组织M4受体表达与分布;其余动物分别提取眼视网膜组织RNA和蛋白质,逆转录聚和酶链反应(RT-PER)、蛋白质免疫印迹(Western blot)法检测M4受体mRNA和蛋白质表达.结果 免疫组织化学染色显示,M4受体分布于豚鼠视网膜RPE细胞层,光感受器细胞层、外核层、内核层,大多数神经节细胞,以及内从状层和内核层交界区;豚鼠视网膜组织表达M4受体mRNA,扩增产物约为221 bp,阴性对照未见扩增产物.免疫印迹显示,豚鼠视网膜组织表达明显的M4受体免疫反应活性,分子量约为38 000.阴性对照组无阳性表达条带.结论 豚鼠视网膜组织有M4受体蛋白表达.

Expression of M4 receptor In normal guinea pigs retina

Objective To detect the expression and distribution of M4 receptor in normal guinea pigs retina. Methods Twenty one normal 4-week-old triad color guinens were selected. Eyes were enucleated. Seven eyes were fixed, paraffin-embeded and cut into sections. Expression and distribution of M4 receptor were detected using immunohisto chemically. The rest eyes were dissected. Tissue of retina were separated. RNA and protein of different tissue were extracted respectively. M4 receptor at mRNA level and protein level were detected by RT-PCR and Western blot methods. Results receptor at mRNA level was detected in normal guineas retina, with the amplification product at 221 bp. No product was detected in negative control. M4 receptor at protein level was detected in normal guineas retina, with signal band at 38 000. No signal was detect in negative control. As detected by immunohistochemistry, M4 receptor was detected in RPE cells, photoreceptor layer, outer nuclear layer, inner nuclear layer, most ganglion cells and the junctional zone of inner plexiform layer and inner nuclear layer. Conclusion M4 receptor was expressed in normal guineas retina.