Watson Claire J, Lies Sarah M, Minich Rebecca R, Tempel Bruce L
Department of Otolaryngology-Head and Neck Surgery, University of Washington, Box 356515, Seattle, WA, 98195, USA,
J Assoc Res Otolaryngol. 2014 Aug;15(4):543-54. doi: 10.1007/s10162-014-0454-z. Epub 2014 May 6.
The plasma membrane Ca(2+) ATPase 2 (PMCA2) is necessary for auditory transduction and serves as the primary Ca(2+) extrusion mechanism in auditory stereocilia bundles. To date, studies examining PMCA2 in auditory function using mutant mice have focused on the phenotype of late adolescent and adult mice. Here, we focus on the changes of PMCA2 in the maturation of auditory sensitivity by comparing auditory responses to RNA and protein expression levels in haploinsufficient PMCA2 and wild-type mice from P16 into adulthood. Auditory sensitivity in wild-type mice improves between P16 and 3 weeks of age, when it becomes stable through adolescence. In haploinsufficient mice, there are frequency-dependent loss of sensitivity and subsequent recovery of thresholds between P16 and adulthood. RNA analysis demonstrates that α-Atp2b2 transcript levels increase in both wild-type and heterozygous cochleae between P16 and 5 weeks. The increases reported for the α-Atp2b2 transcript type during this stage in development support the requisite usage of this transcript for mature auditory transduction. PMCA2 expression also increases in wild-type cochleae between P16 and 5 weeks suggesting that this critical auditory protein may be involved in normal maturation of auditory sensitivity after the onset of hearing. We also characterize expression levels of two long noncoding RNA genes, Gm15082 (lnc82) and Gm15083 (lnc83), which are transcribed on the opposite strand in the 5' region of Atp2b2 and propose that the lnc83 transcript may be involved in regulating α-Atp2b2 expression.
质膜Ca(2+)ATP酶2(PMCA2)对于听觉转导是必需的,并且是听觉静纤毛束中主要的Ca(2+)排出机制。迄今为止,使用突变小鼠研究PMCA2在听觉功能中的作用主要集中在青少年晚期和成年小鼠的表型上。在这里,我们通过比较单倍体不足的PMCA2小鼠和野生型小鼠从P16到成年期的听觉反应与RNA和蛋白质表达水平,来关注PMCA2在听觉敏感性成熟过程中的变化。野生型小鼠的听觉敏感性在P16和3周龄之间提高,之后在青春期保持稳定。在单倍体不足的小鼠中,在P16到成年期之间存在频率依赖性的敏感性丧失以及随后阈值的恢复。RNA分析表明,在P16和5周之间,野生型和杂合子耳蜗中的α-Atp2b2转录水平均增加。在此发育阶段报道的α-Atp2b2转录本类型的增加支持了该转录本在成熟听觉转导中的必要用途。在P16和5周之间,野生型耳蜗中的PMCA2表达也增加,这表明这种关键的听觉蛋白可能参与听力开始后听觉敏感性的正常成熟。我们还对两个长链非编码RNA基因Gm15082(lnc82)和Gm15083(lnc83)的表达水平进行了表征,它们在Atp2b2的5'区域的相反链上转录,并提出lnc83转录本可能参与调节α-Atp2b2的表达。