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目的研究大鼠CNS肾母细胞瘤过度表达基因(nov)mRNA神经元的发育。方法原位杂交组织化学和逆转录PCR。结果P0~P4:原位杂交方法最早检测到novmRNA神经元是P0,P0~P4阳性信号均较弱,分布于脑桥和脊髓等。P5:丘脑部分核团和海马锥体细胞层观察到阳性神经元。P8:novmRNA神经元数目增多,边缘系统及丘脑明显阳性;大脑皮质和下丘脑阳性较弱。P15:丘脑、下丘脑和大脑皮质novmRNA神经元增多,其余核团稳定在P8水平。P30:在脑干观察到强烈的阳性信号,大脑皮质强阳性,海马、丘脑和下丘脑标记较强。P60:脑桥和延髓的标记仍然很强,大脑皮质、丘脑和下丘脑的阳性信号稳定在P30水平。P300:novmRNA神经元显著减少,阳性信号明显减弱。逆转录PCR检测结果与原位杂交所得结果基本相符,在出生前2d检测到novmR-NA,其表达高峰出现在P30~P60之间,随之下降,P150和P300表达弱。结论nov基因与其他早期反应基因不同,其表达高峰不是在胚胎发育早期,而是在近性成熟时期,提示nov基因主要在神经系统发育、分化及功能活动中起作用。
Objective To study the development of rat neuronal neoplasia overexpressing gene (nov) mRNA. Methods In situ hybridization and reverse transcription PCR. Results P0 ~ P4: The earliest detected nov mRNA neurons in situ hybridization were P0, P0 ~ P4 positive signals were weaker, located in the pons and spinal cord. P5: Positive neurons were observed in thalamic nuclei and hippocampal pyramidal cells. P8: novmRNA neurons increased, the edge of the system and the thalamus was significantly positive; cerebral cortex and hypothalamus is weakly positive. P15: NovmRNA neurons in the thalamus, hypothalamus, and cerebral cortex increased while the remaining nuclei stabilized at the P8 level. P30: A strong positive signal was observed in the brainstem, strongly positive in the cerebral cortex, and marked in the hippocampus, thalamus and hypothalamus. P60: Pons and medulla marked still strong, positive signals in cerebral cortex, thalamus and hypothalamus stabilized at P30 level. P300: novmRNA neurons decreased significantly, positive signals decreased significantly. Reverse transcriptase PCR results were in good agreement with those obtained by in situ hybridization. NovmR-NA was detected at 2 days before birth, and peaked at P30-P60, followed by a decrease, while P150 and P300 were weakly expressed. Conclusion The nov gene is different from other early response genes. The peak of expression of the nov gene is not in the early embryonic stage, but in the near mature stage, suggesting that the nov gene mainly plays a role in the development, differentiation and functional activity of the nervous system.