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

Patricia A. Castillo, Eric B. Nonnecke, Daniel T. Ossorio, Michelle T.N. Tran, Stephanie M. Goley, Bo Lönnerdal, Mark Underwood, Charles L Bevins

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Number of pages1
JournalScientific reports
Volume9
Issue number1
DOIs
StatePublished - Sep 11 2019

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Paneth Cells
Defensins
Inbred C57BL Mouse
Messenger RNA
Peptides
Small Intestine
DNA Primers
Mammals
Plasmids
Homeostasis
Complementary DNA
Polymerase Chain Reaction

ASJC Scopus subject areas

  • General

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An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice. / Castillo, Patricia A.; Nonnecke, Eric B.; Ossorio, Daniel T.; Tran, Michelle T.N.; Goley, Stephanie M.; Lönnerdal, Bo; Underwood, Mark; Bevins, Charles L.

In: Scientific reports, Vol. 9, No. 1, 11.09.2019.

Research output: Contribution to journalArticle

Castillo, Patricia A. ; Nonnecke, Eric B. ; Ossorio, Daniel T. ; Tran, Michelle T.N. ; Goley, Stephanie M. ; Lönnerdal, Bo ; Underwood, Mark ; Bevins, Charles L. / An Experimental Approach to Rigorously Assess Paneth Cell α-Defensin (Defa) mRNA Expression in C57BL/6 Mice. In: Scientific reports. 2019 ; Vol. 9, No. 1.
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