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VGLUT3 在大鼠耳蜗中的时空表达和细胞定位。

Temporospatial expression and cellular localization of VGLUT3 in the rat cochlea.

机构信息

Department of Otolaryngology-Head and Neck Surgery, Key Laboratory of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, China.

出版信息

Brain Res. 2013 Nov 6;1537:100-10. doi: 10.1016/j.brainres.2013.09.019. Epub 2013 Sep 21.

Abstract

Vesicular glutamate transporter 3 (VGLUT3) plays an important role in hearing, and VGLUT3 knockout mice are deaf. However, the mechanisms whereby VGLUT3 exerts its effects in the cochlea are not well established. Elucidating the developmental and aging dynamics of VGLUT3 localization and expression in the cochlea would aid a functional understanding of auditory glutamatergic transmission. In this study, we characterized the expression of VGLUT3 in rat auditory epithelium and spiral ganglion cells (SGCs) during postnatal development and with age using immunohistochemistry and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). We observed VGLUT3 expression in cochlea inner hair cells (IHCs) and SGCs as early as postnatally, and its expression increased gradually with maturity. However, the expression of VGLUT3 in the IHCs decreased in the aging rats compared to postnatal day 60 (P60), and VGLUT3 was still expressed in the remaining SGCs in the aging rats. VGLUT3-immunopositive punctate structures were only found in the cytoplasm of SGCs, not IHCs, at P7, or later. Variation in VGLUT3 mRNA expression evaluated by quantitative real time RT-PCR was consistent with the immunohistochemical data except for no statistical difference between P60 and aging rats in the whole cochlea level. Our data suggest that VGLUT3 likely has developmental and physiological roles in the rat cochlea during postnatal development as well as later in life. VGLUT3 may have additional unknown roles in SGCs besides glutamate transport.

摘要

囊泡谷氨酸转运体 3(VGLUT3)在听力中发挥着重要作用,VGLUT3 敲除小鼠耳聋。然而,VGLUT3 在耳蜗中发挥作用的机制尚未完全确定。阐明 VGLUT3 在耳蜗中的定位和表达的发育和衰老动态将有助于理解听觉谷氨酸能传递的功能。在这项研究中,我们使用免疫组织化学和实时定量逆转录聚合酶链反应(RT-PCR)技术,研究了 VGLUT3 在大鼠听觉上皮和螺旋神经节细胞(SGCs)中的表达在出生后发育和衰老过程中的特征。我们观察到 VGLUT3 在耳蜗内毛细胞(IHC)和 SGCs 中的表达早在出生后就出现了,并随着成熟逐渐增加。然而,与 P60 相比,衰老大鼠耳蜗中的 IHC 中 VGLUT3 的表达减少,而 VGLUT3 在衰老大鼠中剩余的 SGCs 中仍有表达。VGLUT3 免疫阳性点状结构仅在 P7 或之后的 SGC 细胞质中发现,而不在 IHC 中发现。通过实时定量 RT-PCR 评估的 VGLUT3 mRNA 表达变化与免疫组织化学数据一致,除了整个耳蜗水平上 P60 和衰老大鼠之间没有统计学差异。我们的数据表明,VGLUT3 在大鼠耳蜗的出生后发育以及以后的生命中可能具有发育和生理作用。VGLUT3 除了谷氨酸转运之外,在 SGC 中可能还有其他未知的作用。

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