Abstract

The small intestine of the piglet has evolved to be permeable immediately after birth to facilitate the uptake of colostrum-derived immunoglobulins as well as other macromolecules, and cells. However, the precise timing of gut closure in today’s precocious pig is not known. We gavaged piglets immediately after birth and at 1-h after birth with Cy5-labeled Ovalbumin (Cy5-Ova) then harvested their small intestine’s 6–7 h later. To assess localization of Cy5-Ova in the small intestinal epithelial cells, we performed immunohistochemistry using a basolateral surface marker and a recycling endosome marker called pIgR, the late endosomal marker Rab7, and the lysosomal marker LAMP-1. Cy5-Ova co-localized with Rab7 and LAMP-1 in the duodenum and jejunum of 0-h old and 1-h old gavaged piglets, but only in the ileum of 0-h gavaged piglets. These data suggest that movement of Cy5-Ova through the late endosomes to the lysosomes was much reduced in the ileum of 1-h gavaged piglets. Cy5-Ova was largely present in epithelial cell digestive and transport vacuoles, but it did not colocalize with pIgR-positive endosomes in 0-h and 1-h gavaged piglets. Differences in macromolecular uptake across the different regions of the small intestine after only 1-h may be due to prior processing of colostral macromolecules, changes in the intestine due to initiation of colonization by microflora and/or the initiation of gut-closure. Understanding the relationship between the localization of Cy5-Ova and small intestinal permeability may contribute to establishing whether oral vaccination in the newborn can capitalize on the transient permeability before gut closure to promote immune protection.

Highlights

  • Fetal and newborn piglet intestinal enterocytes possess an apical canalicular system which allows for the production of cytoplasmic vacuoles of various sizes, which are vital for colostrum uptake (Skrzypek et al 2018)

  • Cy5-labeled Ovalbumin (Cy5-Ova) colocalized with lysosomal marker Lysosomal-associated membrane protein 1 (LAMP-1) in duodenal, jejunal and ileal intestinal epithelial cell (IEC) in 0-h gavaged piglets as well as being present witihin vacuoles

  • Our research shows that in duodenal, jejunal and ileal IECs, Cy5-Ova does not colocalize with pIgR on the surface of the cells or within pIgR + endosomes, regardless of whether they were gavaged pre- or post-suckling

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Summary

Introduction

Fetal and newborn piglet intestinal enterocytes possess an apical canalicular system which allows for the production of cytoplasmic vacuoles of various sizes, which are vital for colostrum uptake (Skrzypek et al 2018). These fetalderived enterocytes have large vacuoles (referred to as vacuolated fetal enterocytes (VFEs) that absorb and transport macromolecules either to the basolateral surface where they are expelled, or to the lysosomes where they are digested (Skrzypek et al 2018, 2007). Transport vacuoles are formed immediately apical to the nucleus of the enterocyte after newborn piglets consume colostrum and the macromolecules are taken up via endocytosis.

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