A combined in situ hybridization and RT-PCR method to detect spatial and temporal patterns of Noggin gene expression in embryonic and postnatal rat hippocampus

Xiaotang Fan , Haiwei Xu , Yunjian Huang , Wenqin Cai
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引用次数: 10

Abstract

Recent studies indicate that Noggin not only plays an important role in the early development of the nervous system, but may also plays a role in postnatal central nervous system (CNS) development. In this study, we examined the relative levels and localization of Noggin mRNA in the hippocampus of rats of different developmental stages with reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization. RT-PCR showed that the peak level of expression of Noggin mRNA was observed at embryonic day 13 (E13), subsequently gradually declined at 1–3 months (P1-3M) postnatal, and was detected only at a low level at P18M. In situ hybridization revealed that at embryonic stages, Noggin mRNA was localized throughout all hippocampal regions, whereas at early postnatal ages, Noggin mRNA was primarily localized in the anterior subiculum. At later postnatal ages, Noggin mRNA expression was obviously observed in the dentate gyrus and in the CA1-CA3 pyramidal cell layers. Taken together, our results demonstrate that Noggin is expressed in embryonic and postnatal hippocampus, and that the temporal and spatial patterns of its expression is developmentally regulated.

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采用原位杂交和RT-PCR相结合的方法检测胚胎和出生后大鼠海马中Noggin基因表达的时空格局
最近的研究表明,Noggin不仅在神经系统的早期发育中起重要作用,而且可能在出生后中枢神经系统(CNS)的发育中起作用。本研究采用逆转录聚合酶链反应(RT-PCR)和原位杂交技术检测了不同发育阶段大鼠海马中Noggin mRNA的相对水平和定位。RT-PCR结果显示,Noggin mRNA的表达量在胚胎第13天(E13)达到峰值,随后在出生后1-3个月(P1-3M)逐渐下降,仅在P18M时低表达。原位杂交显示,在胚胎阶段,Noggin mRNA定位于所有海马区域,而在出生后早期,Noggin mRNA主要定位于前骨下带。出生后较晚,在齿状回和CA1-CA3锥体细胞层中明显观察到Noggin mRNA的表达。综上所述,我们的研究结果表明,Noggin在胚胎和出生后海马中表达,并且其表达的时间和空间模式受发育调节。
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