Du Zhihui, Liu Han, Chu Hanqi, Chen Jin
Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Neurosurgery, Hubei Provincial Hospital of Integrative Medicine, Wuhan, China.
J Int Adv Otol. 2025 Mar 25;21(2):1-6. doi: 10.5152/iao.2025.241581.
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.
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.
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.
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.
尽管已对成年大鼠耳蜗中Cav1.3的多种功能进行了探索,但它在Sprague-Dawley(SD)大鼠发育中的血管纹(SV)中的作用鲜有报道。
将出生后0至21天(p0 - p21)的SD大鼠用作实验模型,并按年龄随机分为4组:p0、p7、p14和p21。使用出生3天的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)和免疫荧光技术在体内鉴定Cav1.3。应用免疫细胞化学方法在体外探索Cav1.3的表达。
定量逆转录聚合酶链反应显示,在包括SV和螺旋韧带的耳蜗外侧壁(STV)中检测到Cav1.3钙通道基因(CACNA1D)。免疫荧光照片显示,Cav1.3在SV中强烈表达,在螺旋韧带和螺旋隆起中弱表达。在SV中,Cav1.3的免疫荧光标记存在于MCs、中间细胞和基底细胞中。免疫细胞化学证实Cav1.3在MCs的细胞膜中表达。此外,在发育中的STV中,Cav1.3在蛋白质和mRNA水平上的表达均增加。
Cav1.3主要存在于出生后SD大鼠SV中MCs的细胞膜中。此外,在SD大鼠的耳蜗外侧壁,包括SV中,Cav1.3蛋白和mRNA随发育而增加。Cav1.3表达的改变可能影响离子稳态,并有利于耳蜗的正常功能和发育。