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目的 研究大鼠N4mRNA神经元在前皮质与海马的时空表达模式。 方法 地高辛标记的cRNA探针原位杂交组织化学和逆转录PCR技术。 结果 E13 E16 :原位杂交方法最早检测到N4mRNA神经元 ,在海马与前皮质呈强阳性信号。P0～P2 :N4mRNA阳性信号在海马开始减弱 ,在前皮质仍维持较高的水平。P7:N4mRNA阳性信号在海马明显减弱 ,仅在海马的齿状回见部分阳性神经元 ,在前皮质的水平轻度减低。P14 :N4mRNA神经元数目增多 ,主要位于海马齿状回的颗粒细胞层 ;P2 1～P2 8:N4mRNA在海马的齿回呈强阳性信号 ,在CA3区呈中等程度的阳性信号 ,CA1区、CA2区及下托可见部分阳性神经元 ,前皮质可见Ⅱ /Ⅲ层的锥体细胞层呈强阳性信号。P90～P15 0 :在海马的齿状回、CA3区 ,CA1区、CA2区及下托见中等强度的N4mRNA阳性信号。P30 0 :N4mRNA神经元在海马与皮层的表达明显减少 ,仅见部分散在的阳性神经元。逆转录PCR检测结果与原位杂交所得结果基本相符 ,在胚胎期N4mRNA表达最强 ,生后减弱 ,P7降至最低 ,P2 1～P2 8为生后的表达高峰 ,随之下降 ,P30 0表达弱。N4基因在不同发育时期皮层的表达 ,在胚胎期信号最强 ,生后略微减弱 ,组间差异不明显。 结论 N4基因特异的表达于海马 ,并在海马的形成期表达最高 ,可能对海马的传入通路的?
Objective To study the temporal and spatial patterns of N4 mRNA expression in rat anterior cortex and hippocampus. Methods Digoxigenin-labeled cRNA probe was used to detect the expression of c-Jun by reverse transcription polymerase chain reaction (PCR) in situ hybridization. Results E13 E16: The earliest detection of N4 mRNA neurons by in situ hybridization showed a strong positive signal in the hippocampus and the anterior cortex. P0 ~ P2: N4 mRNA positive signal in the hippocampus began to weaken, the anterior cortex is still maintained at a high level. P7: N4 mRNA positive signal was significantly weakened in the hippocampus, and only partially positive neurons were seen in the dentate gyrus of the hippocampus, with a slight decrease in the level of the anterior cortex. P14: The number of N4mRNA neurons increased, mainly located in the granular cell layer of hippocampal dentate gyrus; P2 1 ~ P2 8: N4mRNA showed a strong positive signal in the hippocampus and a moderate positive signal in the CA3 area. CA1, CA2 Under the area and the next part of visible positive neurons, the anterior cortex shows Ⅱ / Ⅲ pyramidal cell layer was strongly positive signal. P90 ~ P15 0: in the hippocampus dentate gyrus, CA3 area, CA1 area, CA2 area and under the care to see moderate-intensity N4 mRNA positive signals. P30 0: The expression of N4 mRNA in hippocampus and cortex decreased significantly, only some scattered positive neurons were observed. The results of RT-PCR were basically consistent with those obtained by in situ hybridization. The expression of N4 mRNA was the strongest in embryonic stage, but decreased in P7 after birth, and P7 decreased to the lowest level. weak. N4 gene in different developmental stages of the cortex, the strongest signal in the embryo, slightly weakened after birth, the difference was not obvious. Conclusion The N4 gene is specifically expressed in the hippocampus and expressed the highest in the hippocampus during formation and may be involved in the afferent pathway of the hippocampus.