HTR4 gene structure and altered expression in the developing lung

Hodge, Emily; Nelson, Carl P.; Miller, Suzanne; Billington, Charlotte K.; Stewart, Ceri E.; Swan, Caroline; Malarstig, Anders; Henry, Amanda P.; Gowland, Catherine; Mel�n, Erik; Hall, Ian P.; Sayers, Ian

Abstract

Background: Meta-analyses of genome-wide association studies (GWAS) have identified single nucleotide
polymorphisms (SNPs) spanning the 5-hydroxytryptamine receptor 4 (5-HT4R) gene (HTR4) associated with lung
function. The aims of this study were to i) investigate the expression profile of HTR4 in adult and fetal lung tissue
and cultured airway cells, ii) further define HTR4 gene structure and iii) explore the potential functional implications of key SNPs using a bioinformatic approach.

Methods: Following reverse transcription (RT)-PCR in human brain, 5′ rapid amplification of cDNA ends (5′ RACE) was used to examine the exonic structure of HTR4 at the 5′ end. Quantitative (Q)-PCR was used to quantify HTR4 mRNA expression in total RNA from cultured airway cells and whole lung tissue. Publically available gene microarray data on fetal samples of estimated gestational age 7–22 weeks were mined for HTR4 expression. Immunohistochemistry (IHC; in adult and fetal lung tissue) and a radioligand binding assay (in cultured airway cells) were used to analyze 5HT4R protein expression.

Results: IHC in adult lung, irrespective of the presence of chronic obstructive pulmonary disease (COPD), suggested low
level expression of 5-HT4R protein, which was most prominent in alveolar pneumocytes. There was evidence of differential 5-HT4R protein levels during gestation in fetal lung, which was also evident in gene expression microarray data. HTR4 mRNA expression, assessed by Q-PCR, was

Citation

Hodge, E., Nelson, C. P., Miller, S., Billington, C. K., Stewart, C. E., Swan, C., …Sayers, I. (2013). HTR4 gene structure and altered expression in the developing lung. Respiratory Research, 14(77), https://doi.org/10.1186/1465-9921-14-77

• Journal Article Type: Article
• Publication Date: Jul 26, 2013
• Deposit Date: May 2, 2014
• Publicly Available Date: May 2, 2014
• Journal: Respiratory Research
• Print ISSN: 1465-9921
• Electronic ISSN: 1465-9921
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 14
• Issue: 77
• DOI: https://doi.org/10.1186/1465-9921-14-77
• Public URL: https://nottingham-repository.worktribe.com/output/716126
• Publisher URL: http://respiratory-research.com/content/14/1/77