Huang Yibo, Chi Fanglu, Han Zhao, Yang Juanmei, Gao Wenyuan, Li Yike
Department of Otorhinolaryngology, Eye, Ear, Nose and Throat Hospital, Fudan University, 83 Fen Yang Road, Shanghai 200031, People's Republic of China.
Brain Res. 2009 Jun 18;1276:31-8. doi: 10.1016/j.brainres.2009.04.036. Epub 2009 May 3.
Overexpression of Math1 has been demonstrated to induce robust ectopic new hair cells in greater epithelial ridge (GER) and lesser epithelial ridge (LER) of the postnatal rats' cochlear in vitro. In spite of the similarities between cochlear and vestibular epithelia in origin and structure, no similar results have been reported in the non-sensory region of vestibular epithelia in vitro. In the study, the adenoviral vectors inserted with Math1 gene were constructed to examine their effect on vestibular epithelia in postnatal rats. In vivo, the adenovirus vectors administered in vestibular perilymphatic or endolymphatic space, their transduction efficiency and other indexes are different. We set up a culture construction to simulate and show the differences. In the study, we also developed a new dissection protocol to be quick to harvest the whole maculae and cristae. The new ectopic vestibular hair cell-like cells induced by overexpression of Math1, were found to appear in the non-sensory region of the postnatal rats' vestibular epithelia as observed in the cochlear, and the number of the new cells was different when a different virus administration was simulated in vestibular perilymphatic or endolymphatic space, suggesting that the cells found could have the capability to differentiate into new hair cells. Our study might pave the way for further in vivo studies on vestibular Math1 adenoviral vector gene transfer.
已证明Math1的过表达可在体外诱导出生后大鼠耳蜗的大上皮嵴(GER)和小上皮嵴(LER)中产生大量异位新毛细胞。尽管耳蜗上皮和前庭上皮在起源和结构上有相似之处,但体外前庭上皮的非感觉区域尚未有类似结果的报道。在本研究中,构建了插入Math1基因的腺病毒载体,以研究其对出生后大鼠前庭上皮的作用。在体内,将腺病毒载体注入前庭外淋巴间隙或内淋巴间隙,其转导效率和其他指标有所不同。我们建立了一种培养结构来模拟并展示这些差异。在本研究中,我们还开发了一种新的解剖方法,能够快速获取整个黄斑和嵴。结果发现,Math1过表达诱导产生的新的异位前庭毛细胞样细胞出现在出生后大鼠前庭上皮的非感觉区域,这与耳蜗中的情况类似,并且在前庭外淋巴间隙或内淋巴间隙模拟不同的病毒给药时,新细胞的数量有所不同,这表明所发现的细胞可能具有分化为新毛细胞的能力。我们的研究可能为进一步开展关于前庭Math1腺病毒载体基因转移的体内研究铺平道路。