Kerschner Joseph E, Meyer Tanya K, Burrows Amy, Ehrlich Garth, Post J Christopher
Division of Pediatric Otolaryngology, Medical College of Wisconsin, 9000 W. Wisconsin Avenue, Milwaukee, WI 53226, USA.
Int J Pediatr Otorhinolaryngol. 2006 Aug;70(8):1449-56. doi: 10.1016/j.ijporl.2006.03.007. Epub 2006 May 2.
To identify mucin genes in chinchilla middle ear epithelium and characterize complimentary deoxyribonucleic acid (cDNA) sequences to facilitate further investigations into mucin physiology and pathophysiology on a molecular level using the chinchilla model.
Chinchilla mucin gene exploration and cDNA characterization was accomplished using reverse transcriptase-polymerase chain reactions (RT-PCR). Forward and reverse primer pairs were designed using consensus sequences available for human and rodent species. Chinchilla middle ear epithelium was harvested and primary cell cultures (CMEEC) were established. The CMEEC were explored for the expression of chinchilla mucin genes 1, 2, 4 and 5AC (cMuc1, cMuc2, cMuc4 and cMuc5AC). Identified cDNA amplicons for each of these genes was sequenced and homology compared to previously published human and rodent sequences.
CMEEC express all four of the mucin genes cMuc1, cMuc2, cMuc4 and cMuc5AC. cDNA amplicons for each of the genes were able to be sequenced with lengths ranging from 66 to 362 base pairs. Each of the chinchilla cDNA sequences expressed significant homology with published human and rodent cDNA for these mucin genes. A cDNA sequence for the housekeeping gene, beta-actin, was also identified.
Chinchilla middle ear epithelium grown in culture expresses the mucin genes 1, 2, 4 and 5AC, which have been identified as important in mucin regulation in the middle ear. cDNA sequences corresponding to these mucin genes were identified and may serve as important molecular tools in future studies of otitis media using the chinchilla model.
鉴定毛丝鼠中耳上皮中的黏蛋白基因，并对互补脱氧核糖核酸（cDNA）序列进行特征分析，以便利用毛丝鼠模型在分子水平上进一步研究黏蛋白的生理和病理生理学。
采用逆转录聚合酶链反应（RT-PCR）完成毛丝鼠黏蛋白基因探索和cDNA特征分析。使用人类和啮齿动物物种可用的共有序列设计正向和反向引物对。采集毛丝鼠中耳上皮并建立原代细胞培养（CMEEC）。对CMEEC进行毛丝鼠黏蛋白基因1、2、4和5AC（cMuc1、cMuc2、cMuc4和cMuc5AC）表达情况的探索。对这些基因各自鉴定出的cDNA扩增子进行测序，并与先前发表的人类和啮齿动物序列进行同源性比较。
CMEEC表达所有四种黏蛋白基因cMucl、cMuc2、cMuc4和cMuc5AC。每个基因的cDNA扩增子都能够测序，长度范围为66至362个碱基对。这些毛丝鼠cDNA序列中的每一个与已发表的这些黏蛋白基因的人类和啮齿动物cDNA都表现出显著同源性。还鉴定出管家基因β-肌动蛋白的一个cDNA序列。
培养的毛丝鼠中耳上皮表达黏蛋白基因1、2、4和5AC，这些基因已被确定在中耳黏蛋白调节中很重要。鉴定出了与这些黏蛋白基因相对应的cDNA序列，它们可能成为未来利用毛丝鼠模型研究中耳炎时的重要分子工具。
