Cav1.3在大鼠血管纹发育中的表达模式。

Zhihui Du, Han Liu, Hanqi Chu, Jin Chen
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引用次数: 0

摘要

背景:成年大鼠耳蜗中Cav1.3的多种功能已被探索,但其在SD大鼠血管纹(SV)发育中的作用却鲜有报道。方法:取出生后0 ~ 21 d (p0 ~ p21)的Sprague-Dawley大鼠作为实验模型,按年龄随机分为p0、p7、p14、p21组。p3 SD大鼠培养原代边缘细胞(MCs)。采用定量逆转录聚合酶链反应(qRT-PCR)和免疫荧光技术对Cav1.3进行体内鉴定。采用免疫细胞化学方法探讨Cav1.3在体外的表达。结果:定量逆转录聚合酶链反应显示,在耳蜗侧壁(STV)中检测到Cav1.3钙通道基因(CACNA1D),包括SV和螺旋韧带。免疫荧光图像显示,Cav1.3在SV中表达强烈,在螺旋韧带和螺旋突起中表达弱。在SV中,MCs、中间细胞和基底细胞中存在Cav1.3的免疫荧光标记。免疫细胞化学证实Cav1.3在MCs细胞膜中表达。此外,Cav1.3在发育性STV中的表达在蛋白和mRNA水平上均有所增加。结论:Cav1.3主要存在于出生后SD大鼠SV的MCs细胞膜中。此外,在SD大鼠耳蜗外侧壁(包括SV)中,Cav1.3蛋白和mRNA随着发育而增加。Cav1.3表达的改变可能影响离子稳态,有利于耳蜗的正常功能和发育。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Expression Patterns of Cav1.3 in the Developing Stria Vascularis of Sprague-Dawley Rats.

Background: Although multiple functions of Cav1.3 in adult rat cochlea have been explored, its role in developing stria vascularis (SV) of Sprague-Dawley (SD) rats has rarely been reported.

Methods: Sprague-Dawley rats from postnatal 0 to 21 days (p0-p21) were utilized for the experimental model and classified into 4 groups by age randomly: p0, p7, p14, and p21. p3 SD rats were employed to culture primary marginal cells (MCs). The quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunofluorescence technique were employed to identify Cav1.3 in vivo. Immunocytochemistry was applied to explore the expression of Cav1.3 in vitro.

Results: Quantitative reverse transcription polymerase chain reaction showed that the Cav1.3 calcium channel gene (CACNA1D) was measured in the cochlear lateral wall (STV), which included SV and spiral ligament. Immunofluorescence photographs revealed that Cav1.3 was intensively expressed in the SV, weakly expressed in the spiral ligament and spiral prominence. In the SV, immunofluorescence labeling of Cav1.3 was present in MCs, intermediate cells, and basal cells. Immunocytochemistry confirmed that Cav1.3 was expressed in the cytomembrane of the MCs. Moreover, the expression of Cav1.3 was increased in the developmental STV at both the protein and mRNA levels.

Conclusion: Cav1.3 was mainly present in the cytomembrane of MCs in the SV of postnatal SD rats. Additionally, Cav1.3 protein and mRNA increased with development in the cochlear lateral wall of SD rats, including in the SV. The alteration of Cav1.3 expression may influence the homeostasis of ions and benefit the normal function and development of the cochlea.

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