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Analysis of Mir-9 Expression Pattern in Rat Retina During Postnatal Development

International journal of molecular sciences(2021)SCI 2区SCI 3区

Univ Pecs | Janos Szentagothai Res Ctr

Cited 4|Views29
Abstract
It is well established that miR-9 contributes to retinal neurogenesis. However, little is known about its presence and effects in the postnatal period. To expand our knowledge, miRNA-small RNA sequencing and in situ hybridization supported by RT-qPCR measurement were carried out. Mir-9 expression showed two peaks in the first three postnatal weeks in Wistar rats. The first peak was detected at postnatal Day 3 (P3) and the second at P10, then the expression gradually decreased until P21. Furthermore, we performed in silico prediction and established that miR-9 targets OneCut2 or synaptotagmin-17. Another two microRNAs (mir-135, mir-218) were found from databases which also target these proteins. They showed a similar tendency to mir-9; their lowest expression was at P7 and afterwards, they showed increase. We revealed that miR-9 is localized mainly in the inner retina. Labeling was observed in ganglion and amacrine cells. Additionally, horizontal cells were also marked. By dual miRNA-in situ hybridization/immunocytochemistry and qPCR, we revealed alterations in their temporal and spatial expression. Our results shed light on the significance of mir-9 regulation during the first three postnatal weeks in rat retina and suggest that miRNA could act on their targets in a stage-specific manner.
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mir-9,retina,postnatal development,in situ hybridization,qPCR,next-generation sequencing,synaptotagmin-17,OneCut2
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要点】:该论文通过实验揭示了miR-9在大鼠视网膜出生后发育期间表达模式,发现其在出生后前三周内表达出现两个高峰,并确定了miR-9的潜在靶标OneCut2和synaptotagmin-17,及其在视网膜内层的主要定位。

方法】:采用miRNA小RNA测序、原位杂交以及逆转录定量PCR(RT-qPCR)测量技术来研究miR-9的表达模式。

实验】:在Wistar大鼠中,通过原位杂交和免疫细胞化学技术结合qPCR,研究了miR-9及其同家族成员在视网膜中的时空表达变化,并使用数据库中预测的靶标进行验证。结果显示miR-9在出生后3天(P3)和10天(P10)出现表达高峰,随后逐渐降低至21天(P21)。