An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice

Abstract Abundant evidence from many laboratories supports the premise that α-defensin peptides secreted from Paneth cells are key mediators of host-microbe interactions in the small intestine that contribute to host defense and homeostasis. α-defensins are among the most highly expressed antimicrob...

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Autores principales:	Patricia A. Castillo, Eric B. Nonnecke, Daniel T. Ossorio, Michelle T. N. Tran, Stephanie M. Goley, Bo Lönnerdal, Mark A. Underwood, Charles L. Bevins
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2ee908382b004aaf9bc36a2914fbd87d
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spelling	oai:doaj.org-article:2ee908382b004aaf9bc36a2914fbd87d2021-12-02T15:09:25ZAn Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice10.1038/s41598-019-49471-92045-2322https://doaj.org/article/2ee908382b004aaf9bc36a2914fbd87d2019-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-49471-9https://doaj.org/toc/2045-2322Abstract Abundant evidence from many laboratories supports the premise that α-defensin peptides secreted from Paneth cells are key mediators of host-microbe interactions in the small intestine that contribute to host defense and homeostasis. α-defensins are among the most highly expressed antimicrobial peptides at this mucosal surface in many mammals, including humans and mice; however, there is striking variation among species in the number and primary structure of α-defensin paralogs. Studies of these biomolecules in vivo are further complicated by striking variations between laboratory mouse strains. Herein, we report an experimental approach to determine with precision and specificity expression levels of α-defensin (Defa) mRNA in the small intestine of C57BL/6 mice through an optimized set of oligonucleotide primers for qRT-PCR assays and cloned cDNA plasmids corresponding to the Defa paralogs. This approach demonstrated marked differences in α-defensin expression in C57BL/6 mice with respect to proximal/distal anatomical location and developmental stage, which have not been described previously. These data underscore the importance of careful attention to method (primer choice, proximal vs. distal location, and developmental stage) in analysis of antimicrobial peptide expression and their impact.Patricia A. CastilloEric B. NonneckeDaniel T. OssorioMichelle T. N. TranStephanie M. GoleyBo LönnerdalMark A. UnderwoodCharles L. BevinsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-14 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Patricia A. Castillo Eric B. Nonnecke Daniel T. Ossorio Michelle T. N. Tran Stephanie M. Goley Bo Lönnerdal Mark A. Underwood Charles L. Bevins An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
description	Abstract Abundant evidence from many laboratories supports the premise that α-defensin peptides secreted from Paneth cells are key mediators of host-microbe interactions in the small intestine that contribute to host defense and homeostasis. α-defensins are among the most highly expressed antimicrobial peptides at this mucosal surface in many mammals, including humans and mice; however, there is striking variation among species in the number and primary structure of α-defensin paralogs. Studies of these biomolecules in vivo are further complicated by striking variations between laboratory mouse strains. Herein, we report an experimental approach to determine with precision and specificity expression levels of α-defensin (Defa) mRNA in the small intestine of C57BL/6 mice through an optimized set of oligonucleotide primers for qRT-PCR assays and cloned cDNA plasmids corresponding to the Defa paralogs. This approach demonstrated marked differences in α-defensin expression in C57BL/6 mice with respect to proximal/distal anatomical location and developmental stage, which have not been described previously. These data underscore the importance of careful attention to method (primer choice, proximal vs. distal location, and developmental stage) in analysis of antimicrobial peptide expression and their impact.
format	article
author	Patricia A. Castillo Eric B. Nonnecke Daniel T. Ossorio Michelle T. N. Tran Stephanie M. Goley Bo Lönnerdal Mark A. Underwood Charles L. Bevins
author_facet	Patricia A. Castillo Eric B. Nonnecke Daniel T. Ossorio Michelle T. N. Tran Stephanie M. Goley Bo Lönnerdal Mark A. Underwood Charles L. Bevins
author_sort	Patricia A. Castillo
title	An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
title_short	An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
title_full	An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
title_fullStr	An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
title_full_unstemmed	An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice
title_sort	experimental approach to rigorously assess paneth cell α-defensin (defa) mrna expression in c57bl/6 mice
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/2ee908382b004aaf9bc36a2914fbd87d
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An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice

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