Dataset Information

Extending the chinchilla middle ear epithelial model for mucin gene investigation.

ABSTRACT:

Objectives

To investigate the expression of recently identified human mucin genes in an in vivo model of the chinchilla middle ear epithelium (CMEE).

Methods

CMEE was harvested, RNA was extracted and primers were designed for RT-PCR to assess for expression of mucin genes Muc6, Muc17 and Muc18. Further sequencing of these genes was also accomplished.

Results

Mucin genes Muc6, Muc17 and Muc18 was assessed and found to be identical to the expression in human and mouse MEE.

Conclusion

This study further characterizes mucin gene expression in the CMEE and provides additional sequence data for chinchilla middle ear genes. The concordance of this gene expression data to that of both the human and mouse models further demonstrates the utility of this animal model in OM investigations.

SUBMITTER: Kerschner JE

PROVIDER: S-EPMC2922454 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Extending the chinchilla middle ear epithelial model for mucin gene investigation.

Kerschner Joseph E JE Khampang P P Samuels Tina T

International journal of pediatric otorhinolaryngology 20100629 9

<h4>Objectives</h4>To investigate the expression of recently identified human mucin genes in an in vivo model of the chinchilla middle ear epithelium (CMEE).<h4>Methods</h4>CMEE was harvested, RNA was extracted and primers were designed for RT-PCR to assess for expression of mucin genes Muc6, Muc17 and Muc18. Further sequencing of these genes was also accomplished.<h4>Results</h4>Mucin genes Muc6, Muc17 and Muc18 was assessed and found to be identical to the expression in human and mouse MEE.<h4 ...[more]

PMID: 20591507

Similar Datasets

Project description:Haemophilus influenzae is a significant cause of childhood otitis media, and also has an absolute growth requirement for heme. Recent microarray studies using three H. influenzae isolates were used to propose a putative core of genes responsive to iron and heme levels. Included in the core modulon were thirty seven genes that are preferentially expressed under iron/heme limitation, most of which are directly involved with iron and or heme acquisition. In this report, the core iron/heme modulon was further refined following microarray analysis of two additional nontypeable H. influenzae isolates from patients with otitis media. The transcriptional status of the genes comprising the refined iron/heme core modulon was then assessed in vivo, in a chinchilla model of otitis media. These in vivo experiments were performed to address the hypothesis that iron and heme regulated genes are both highly expressed in vivo and important, during clinical infection.Microarray analysis of two additional H. influenzae strains resulted in the definition of a core of iron/heme responsive genes. This core consisted of 35 genes maximally expressed under heme restriction and a further 20 genes maximally expressed in heme replete conditions. In vivo studies were performed with two nontypeable H. influenzae strains, 86-028NP and HI1722. The majority of operons identified as members of the core modulon by microarray were also actively upregulated in the chinchilla ear during otitis media. In 86-028NP, 70% of the operons were significantly upregulated while in HI1722 100% of the operons were upregulated in samples recovered from the chinchilla middle ear.This study elucidates a conserved core of H. influenzae genes the transcription of which is altered by the availability of iron and heme in the growth environment, and further assesses transcription of these genes in vivo. Elucidation of this modulon allows for identification of genes with unrecognized roles in iron/heme acquisition or homeostasis and/or potential roles in virulence. Defining these core genes is also of potential importance in identifying targets for therapeutic and vaccine designs since products of these genes are likely to be preferentially expressed during growth in iron/heme restricted sites of the human body.

Project description:Otitis media (OM) is one of the most globally pervasive pediatric conditions. Translocation of nasopharynx-resident opportunistic pathogens like nontypeable Haemophilus influenzae (NTHi) assimilates into polymicrobial middle ear biofilms, which promote OM pathogenesis and substantially diminish antibiotic efficacy. Oral or tympanostomy tube (TT)-delivered antibiotics remain the standard of care (SOC) despite consequences including secondary infection, dysbiosis, and antimicrobial resistance. Monoclonal antibodies (mAb) against two biofilm-associated structural proteins, NTHi-specific type IV pilus PilA (anti-rsPilA) and protective tip-region epitopes of NTHi integration host factor (anti-tip-chimer), were previously shown to disrupt biofilms and restore antibiotic sensitivity in vitro. However, the additional criterion for clinical relevance includes the absence of consequential microbiome alterations. Here, nine chinchilla cohorts (n = 3/cohort) without disease were established to evaluate whether TT delivery of mAbs disrupted nasopharyngeal or fecal microbiomes relative to SOC-OM antibiotics. Cohort treatments included a 7d regimen of oral amoxicillin-clavulanate (AC) or 2d regimen of TT-delivered mAb, AC, Trimethoprim-sulfamethoxazole (TS), ofloxacin, or saline. Fecal and nasopharyngeal lavage (NPL) samples were collected before and several days post treatment (DPT) for 16S sequencing. While antibiotic-treated cohorts displayed beta-diversity shifts (PERMANOVA, P < 0.05) and reductions in alpha diversity (q < 0.20) relative to baseline, mAb antibodies failed to affect diversity, indicating maintenance of a eubiotic state. Taxonomic and longitudinal analyses showed blooms in opportunistic pathogens (ANCOM) and greater magnitudes of compositional change (P < 0.05) following broad-spectrum antibiotic but not mAb treatments. Collectively, results showed broad-spectrum antibiotics induced significant fecal and nasopharyngeal microbiome disruption regardless of delivery route. Excitingly, biofilm-targeting antibodies had little effect on fecal and nasopharyngeal microbiomes.

Dataset Information

Extending the chinchilla middle ear epithelial model for mucin gene investigation.

Objectives

Methods

Results

Conclusion

Publications

Extending the chinchilla middle ear epithelial model for mucin gene investigation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets