Mucin gene cDNA sequence characterization in chinchilla middle ear epithelium

被引:9
作者
Kerschner, Joseph E.
Meyer, Tanya K.
Burrows, Amy
Ehrlich, Garth
Post, J. Christopher
机构
[1] Med Coll Wisconsin, Div Pediat Otolaryngol, Milwaukee, WI 53226 USA
[2] Med Coll Wisconsin, Dept Otolaryngol & Commun Sci, Milwaukee, WI 53226 USA
[3] Allegheny Singer Res Inst, Ctr Genom Sci, Pittsburgh, PA 15212 USA
[4] Univ Maryland, Sch Med, Dept Otorhinolaryngol, Baltimore, MD 21201 USA
关键词
otitis media; chinchilla; mucin gene; cDNA;
D O I
10.1016/j.ijporl.2006.03.007
中图分类号
R76 [耳鼻咽喉科学];
学科分类号
100213 ;
摘要
Objectives: 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. Methods: 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. Results: 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. Conclusions: 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. (C) 2006 Elsevier Ireland Ltd. All rights reserved.
引用
收藏
页码:1449 / 1456
页数:8
相关论文
共 27 条
[21]  
NAKAMURA A, 1993, ACTA OTO-LARYNGOL, P75
[22]   PRIMARY CULTURE OF CHINCHILLA MIDDLE-EAR EPITHELIUM [J].
NAKAMURA, A ;
LIM, DJ ;
DEMARIA, TF ;
VANBLITTERSWIJK, CA .
ANNALS OF OTOLOGY RHINOLOGY AND LARYNGOLOGY, 1991, 100 (09) :774-782
[23]   Effects of modified pellicles on Candida biofilm formation on acrylic surfaces [J].
Nikawa, H ;
Nishimura, H ;
Hamada, T ;
Yamashiro, H ;
Samaranayake, LP .
MYCOSES, 1999, 42 (1-2) :37-40
[24]   Implications of the AHRQ evidence report on acute otitis media [J].
Rosenfeld, RM ;
Casselbrant, ML ;
Hannley, MT .
OTOLARYNGOLOGY-HEAD AND NECK SURGERY, 2001, 125 (05) :440-448
[25]   Mucin gene expression in the effusions of otitis media with effusion [J].
Takeuchi, K ;
Yagawa, M ;
Ishinaga, H ;
Kishioka, C ;
Harada, T ;
Majima, Y .
INTERNATIONAL JOURNAL OF PEDIATRIC OTORHINOLARYNGOLOGY, 2003, 67 (01) :53-58
[26]  
van der Velden VHJ, 1998, EUR CYTOKINE NETW, V9, P585
[27]  
Zeiher B G, 1996, Curr Opin Pulm Med, V2, P166, DOI 10.1097/00063198-199605000-00002