Immunohistochemical localization of the Na-K-Cl co-transporter (NKCC1) in the gerbil inner ear
Immunohistochemical localization of the Na-K-Cl co-transporter (NKCC1) in the gerbil inner ear
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DOI：
10.1177/002215549704500601
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发表时间：
1997-06-01
影响因子：
3.2
通讯作者：
Schulte, BA
中科院分区：
文献类型：
--
作者：
Crouch, JJ;Sakaguchi, N;Schulte, BA
我们绘制了细胞和亚细胞分布的钠-钾-氯协同转运蛋白（NKCC）在成年沙鼠内耳免疫染色与单克隆抗体（MAb T4）产生的对人类结肠NKCC。在前庭系统中的血管纹和暗细胞的边缘细胞的基底外侧质膜中观察到重免疫标记。耳蜗螺旋韧带和利姆布斯以及前庭神经感觉上皮下层的纤维细胞亚群也以中等至强强度染色，显然沿着它们的整个质膜。由于单克隆抗体T4识别基底外侧分泌（NKCC 1）和顶端吸收（NKCC 2）亚型的共转运蛋白，我们采用逆转录和聚合酶链反应（RT-PCR），以探讨亚型的多样性在内耳组织。使用基于小鼠和人类序列的NKCC 1和NKCC 2亚型特异性PCR引物，在沙鼠内耳中仅检测到NKCC 1的转录本。丰富的NKCC 1的存在下，在基底外侧质膜的前庭边缘和前庭暗细胞证实了从药理学和生理学数据得出的结论。耳蜗和前庭纤维细胞的高度特化亚群中NKCC 1和Na，K-ATP酶的共表达进一步证明了它们在通过毛细胞泄漏或流出到外淋巴中的K+再循环回到内淋巴中的作用，如当前内耳离子稳态模型中所假设的。
We mapped the cellular and subcellular distribution of the Na-K-Cl co-transporter (NKCC) in the adult gerbil inner ear by immunostaining with a monoclonal antibody (MAb T4) generated against human colon NKCC. Heavy immunolabeling was seen in the basolateral plasma membrane of marginal cells in the stria vascularis and dark cells in the vestibular system. Subpopulations of fibrocytes in the cochlear spiral ligament and limbus and underlying the vestibular neurosensory epithelium also stained with moderate to strong intensity, apparently along their entire plasmalemma. Because MAb T4 recognizes both the basolateral secretory (NKCC1) and the apical absorptive (NKCC2) isoforms of the co-transporter, we employed reverse transcription and the polymerase chain reaction (RT-PCR) to explore isoform diversity in inner ear tissues. Using NKCC1 and NKCC2 isoform-specific PCR primers based on mouse and human sequences, only transcripts for NKCC1 were detected in the gerbil inner ear. The presence of abundant NKCC1 in the basolateral plasmalemma of strial marginal and Vestibular dark cells confirms conclusions drawn from pharmacological and physiological data. The co-expression of NKCC1 and Na,K-ATPase in highly specialized subpopulations of cochlear and vestibular fibrocytes provides further evidence for their role in recycling K+ leaked or effluxed through hair cells into perilymph back to endolymph, as postulated in current models of inner ear ion homeostasis.