Yang D, Thalmann I, Thalmann R, Simmons D D
Siebens Hearing Research Center, Central Institute for the Deaf, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Neurobiol. 2004 Mar;58(4):479-92. doi: 10.1002/neu.10289.
The expression of two calcium-binding proteins of the parvalbumin (PV) family, the alpha isoform (alphaPV) and the beta isoform known as oncomodulin (OM), was investigated in the rat cochlea during postnatal development and related to cholinergic efferent innervation. Using RT-PCR analysis, we found that OM expression begins between postnatal day 2 (P2) and P4, and peaks as early as P10, while alphaPV mRNA begins expression before birth and remains highly expressed into the adult period. Both in situ hybridization and immunoreactivity confirm that OM is uniquely expressed by the outer hair cells (OHCs) in the rat cochlea and occurs after efferent innervation along the cochlear spiral between P2 and P4. In contrast to OM expression, alphaPV immunoreactivity is expressed in both inner hair cells (IHCs) and OHCs at birth. Following olivocochlear efferent innervation, OHCs demonstrate weak OM immunoreactivity beginning at P5 and diminished alphaPV immunoreactivity after P10. In organ cultures isolated prior to the efferent innervation of OHCs, OM immunoreactivity failed to develop in OHCs, but alphaPV immunoreactivity remained present in both IHCs and OHCs. In contrast, organ cultures isolated after efferent innervation of OHCs show OHCs with low levels of OM immunoreactivity and high levels of alphaPV immunoreactivity. This study suggests that OM and alphaPV are differentially regulated in OHCs during cochlear development. Our findings further raise the possibility that the expression of PV proteins in OHCs may be influenced by efferent innervation.
在出生后发育阶段的大鼠耳蜗中,研究了小白蛋白(PV)家族的两种钙结合蛋白α亚型(αPV)和被称为癌调蛋白(OM)的β亚型的表达,并将其与胆碱能传出神经支配相关联。通过逆转录聚合酶链反应(RT-PCR)分析,我们发现OM的表达在出生后第2天(P2)至第4天开始,并早在P10时达到峰值,而αPV mRNA在出生前就开始表达,并在成年期一直保持高表达。原位杂交和免疫反应性均证实,OM在大鼠耳蜗的外毛细胞(OHC)中独特表达,且在P2至P4期间沿着耳蜗螺旋的传出神经支配之后出现。与OM表达不同,αPV免疫反应性在出生时在内毛细胞(IHC)和OHC中均有表达。在OHC传出神经支配后,OHC从P5开始表现出较弱的OM免疫反应性,且在P10后αPV免疫反应性降低。在OHC传出神经支配之前分离的器官培养物中,OHC中未出现OM免疫反应性,但αPV免疫反应性在IHC和OHC中均依然存在。相反,在OHC传出神经支配之后分离的器官培养物显示,OHC具有低水平的OM免疫反应性和高水平的αPV免疫反应性。这项研究表明,在耳蜗发育过程中,OM和αPV在OHC中受到不同的调节。我们的研究结果进一步提出了一种可能性,即OHC中PV蛋白的表达可能受传出神经支配的影响。