Journal article
Partial characterization of normal and Haemophilus influenzae-infected mucosal complementary DNA libraries in chinchilla middle ear mucosa
Annals of otology, rhinology & laryngology, v 119(4)
Apr 2010
PMID: 20433028
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We sought to construct and partially characterize complementary DNA (cDNA) libraries prepared from the middle ear mucosa (MEM) of chinchillas to better understand pathogenic aspects of infection and inflammation, particularly with respect to leukotriene biogenesis and response.
Chinchilla MEM was harvested from controls and after middle ear inoculation with nontypeable Haemophilus influenzae. RNA was extracted to generate cDNA libraries. Randomly selected clones were subjected to sequence analysis to characterize the libraries and to provide DNA sequence for phylogenetic analyses. Reverse transcription-polymerase chain reaction of the RNA pools was used to generate cDNA sequences corresponding to genes associated with leukotriene biosynthesis and metabolism.
Sequence analysis of 921 randomly selected clones from the uninfected MEM cDNA library produced approximately 250,000 nucleotides of almost entirely novel sequence data. Searches of the GenBank database with the Basic Local Alignment Search Tool provided for identification of 515 unique genes expressed in the MEM and not previously described in chinchillas. In almost all cases, the chinchilla cDNA sequences displayed much greater homology to human or other primate genes than with rodent species. Genes associated with leukotriene metabolism were present in both normal and infected MEM.
Based on both phylogenetic comparisons and gene expression similarities with humans, chinchilla MEM appears to be an excellent model for the study of middle ear inflammation and infection. The higher degree of sequence similarity between chinchillas and humans compared to chinchillas and rodents was unexpected. The cDNA libraries from normal and infected chinchilla MEM will serve as useful molecular tools in the study of otitis media and should yield important information with respect to middle ear pathogenesis.
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Details
- Title
- Partial characterization of normal and Haemophilus influenzae-infected mucosal complementary DNA libraries in chinchilla middle ear mucosa
- Creators
- Joseph E Kerschner - Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Children's Hospital of Wisconsin, Milwaukee 53226, USAGeza ErdosFen Ze HuAmy BurrowsJoseph CioffiPawjai KhampangMargaret DahlgrenJay HayesRandy KeefeBenjamin JantoJ Christopher PostGarth D Ehrlich
- Publication Details
- Annals of otology, rhinology & laryngology, v 119(4)
- Publisher
- Sage; United States
- Grant note
- R01 DC005659-05 / NIDCD NIH HHS R01 DC007903 / NIDCD NIH HHS DC007903 / NIDCD NIH HHS R01 DC005659 / NIDCD NIH HHS DC02148 / NIDCD NIH HHS R01 DC004173-08 / NIDCD NIH HHS R01 DC004173 / NIDCD NIH HHS K08 DC000192-05 / NIDCD NIH HHS DC00192 / NIDCD NIH HHS F32 DC000192 / NIDCD NIH HHS R01 DC007903-04 / NIDCD NIH HHS R01 DC002148 / NIDCD NIH HHS R01 DC002148-16S1 / NIDCD NIH HHS DC04173 / NIDCD NIH HHS K08 DC000192 / NIDCD NIH HHS DC05659 / NIDCD NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000276976900011
- Scopus ID
- 2-s2.0-77950837313
- Other Identifier
- 991014878367704721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Otorhinolaryngology