Primary culture of salamander intestinal epithelial cells from nests: whole cell Na+ currents induced by valine

Methods are described for isolating the cell nests, subepithelial clusters of germinative cells, from salamander intestinal mucosa and for growing the nests in culture into polarized monolayers of intestinal epithelial cells. Cells were viable in culture for up to 3 wk. The capacity of the monolayer cells to engage in membrane transport was evaluated using the patch-clamp technique in the whole cell mode. L-Valine (25 mM) induced an inward current in small intestinal cells of 25.8 +/- 5.7 pA and depolarized the cell membrane 14.5 +/- 1.6 mV. L-Alanine and L-phenylalanine were similarly effective, whereas D-valine was ineffective. The Km of the transporter for valine was 90 mM. Replacement of bath Na with tris(hydroxymethyl)aminomethane eliminated the inward current induced by valine. The basal (solute-independent) inward current was also reduced by Na+ replacement. Glucose did not induce a Na+ current. In contrast to the effect of valine on small intestinal cells, large intestinal cells were unresponsive to valine. It is concluded that the cultured small intestinal cells possess Na-amino acid but not Na-sugar cotransport. This profile of behavior is characteristic of undifferentiated small intestinal cells. Primary cultures of salamander small intestinal cells should be useful for studying enterocyte function and the developmental biology of the small intestinal mucosa.

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Immunocytochemical localization of ornithine transcarbamylase in rat intestinal mucosa. Light and electron microscopic study.

Journal of Histochemistry & Cytochemistry ◽

10.1177/36.1.3275711 ◽

1988 ◽

Vol 36 (1) ◽

pp. 29-35 ◽

Cited By ~ 11

Author(s):

Y Hamano ◽

H Kodama ◽

M Yanagisawa ◽

Y Haraguchi ◽

M Mori ◽

...

Keyword(s):

Electron Microscopy ◽

Small Intestine ◽

Epithelial Cells ◽

Light Microscopy ◽

Intestinal Mucosa ◽

Intestinal Epithelial Cells ◽

Protein A ◽

Intestinal Epithelial ◽

Electron Microscopic ◽

Small Intestinal

We investigated light and electron microscopic localization of ornithine transcarbamylase (OTC) in rat intestinal mucosa. In the immunoblotting assay of OTC-related protein, a single protein band with a molecular weight of about 36,500 is observed in extracts of liver and small intestinal mucosa but is not observed in those of stomach and large intestine. For light microscopy, tissue slices of the digestive system were embedded in Epon and stained by using anti-bovine OTC rabbit IgG and the immunoenzyme technique. For electron microscopy, slices of these and the liver tissues were embedded in Lowicryl K4M and stained by the protein A-gold technique. By light microscopy, the absorptive epithelial cells of duodenum, jejunum, and ileum stained positively for OTC, but stomach, large intestine, rectum, and propria mucosa of small intestine were not stained. Electron microscopy showed that gold particles representing the antigenic sites for OTC were confined to the mitochondrial matrix of hepatocytes and small intestinal epithelial cells. However, the enzyme was detected in mitochondria of neither liver endothelial cells, submucosal cells of small intestine, nor large intestinal epithelial cells. Labeling density of mitochondria in the absorptive epithelial cells of duodenum, jejunum, and ileum was about half of that in liver cells.

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Faculty Opinions recommendation of Response of small intestinal epithelial cells to acute disruption of cell division through CDC25 deletion.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1157508.617683 ◽

2009 ◽

Author(s):

John Lazo

Keyword(s):

Cell Division ◽

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Small Intestinal

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Enterococcus faecium PNC01 isolated from the intestinal mucosa of chicken as an alternative for antibiotics to reduce feed conversion rate in broiler chickens

Microbial Cell Factories ◽

10.1186/s12934-021-01609-z ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Yang He ◽

Xuan Liu ◽

Yuanyang Dong ◽

Jiaqi Lei ◽

Koichi Ito ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Conversion Rate ◽

Broiler Chickens ◽

Intestinal Epithelial Cells ◽

Relative Length ◽

Intestinal Epithelial ◽

Feed Conversion ◽

Broiler Production ◽

Feed Conversion Rate

Abstract Background The development and utilization of probiotics had many environmental benefits for replacing antibiotics in animal production. Bacteria in the intestinal mucosa have better adhesion to the host intestinal epithelial cells compared to bacteria in the intestinal contents. In this study, lactic acid bacteria were isolated from the intestinal mucosa of broiler chickens and investigated as the substitution to antibiotic in broiler production. Results In addition to acid resistance, high temperature resistance, antimicrobial sensitivity tests, and intestinal epithelial cell adhesion, Enterococcus faecium PNC01 (E. faecium PNC01) was showed to be non-cytotoxic to epithelial cells. Draft genome sequence of E. faecium PNC01 predicted that it synthesized bacteriocin to perform probiotic functions and bacteriocin activity assay showed it inhibited Salmonella typhimurium from invading intestinal epithelial cells. Diet supplemented with E. faecium PNC01 increased the ileal villus height and crypt depth in broiler chickens, reduced the relative length of the cecum at day 21, and reduced the relative length of jejunum and ileum at day 42. Diet supplemented with E. faecium PNC01 increased the relative abundance of Firmicutes and Lactobacillus, decreased the relative abundance of Bacteroides in the cecal microbiota. Conclusion E. faecium PNC01 replaced antibiotics to reduce the feed conversion rate. Furthermore, E. faecium PNC01 improved intestinal morphology and altered the composition of microbiota in the cecum to reduce feed conversion rate. Thus, it can be used as an alternative for antibiotics in broiler production to avoid the adverse impact of antibiotics by altering the gut microbiota. Graphic Abstract

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miRNA-30e regulates abnormal differentiation of small intestinal epithelial cells in diabetic mice by downregulating Dll4 expression

Cell Proliferation ◽

10.1111/cpr.12230 ◽

2016 ◽

Vol 49 (1) ◽

pp. 102-114 ◽

Cited By ~ 21

Author(s):

Ti-Dong Shan ◽

Hui Ouyang ◽

Tao Yu ◽

Jie-Yao Li ◽

Can-Ze Huang ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Diabetic Mice ◽

Small Intestinal

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Proteomic response of inflammatory stimulated intestinal epithelial cells to in vitro digested plums and cabbages rich in carotenoids and polyphenols

Food & Function ◽

10.1039/c6fo00674d ◽

2016 ◽

Vol 7 (10) ◽

pp. 4388-4399 ◽

Cited By ~ 4

Author(s):

Anouk Kaulmann ◽

Sébastien Planchon ◽

Jenny Renaut ◽

Yves-Jacques Schneider ◽

Lucien Hoffmann ◽

...

Keyword(s):

Epithelial Cells ◽

Inflammatory Bowel Diseases ◽

Intestinal Epithelial Cells ◽

Intestinal Cells ◽

Intestinal Epithelial ◽

Bowel Diseases ◽

Inflammatory Bowel ◽

Proteomic Response

Proteomic response of intestinal cells as a model of inflammatory bowel diseases to digested plum and cabbage rich in polyphenols and carotenoids.

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Mitochondrial gene expression in small intestinal epithelial cells

Biochemical Journal ◽

10.1042/bj3080665 ◽

1995 ◽

Vol 308 (2) ◽

pp. 665-671 ◽

Cited By ~ 5

Author(s):

T P Mayall ◽

I Bjarnason ◽

U Y Khoo ◽

T J Peters ◽

A J S Macpherson

Keyword(s):

Epithelial Cells ◽

Cytochrome Oxidase ◽

Cytochrome B ◽

Intestinal Epithelial Cells ◽

Mitochondrial Gene ◽

Cytochrome Oxidase I ◽

Mrna Levels ◽

Intestinal Epithelial ◽

Mitochondrial Transcription Factor ◽

Small Intestinal

Most mitochondrial genes are transcribed as a single large transcript from the heavy strand of mitochondrial DNA, and are subsequently processed into the proximal mitochondrial (mt) 12 S and 16 S rRNAs, and the more distal tRNAs and mRNAs. We have shown that in intestinal epithelial biopsies the steady-state levels of mt 12 S and 16 S rRNA are an order of magnitude greater than those of mt mRNAs. Fractionation of rat small intestinal epithelial cells on the basis of their maturity has shown that the greatest ratios of 12 S mt rRNA/cytochrome b mt mRNA or 12 S mt rRNA/cytochrome oxidase I mt mRNA are found in the surface mature enterocytes, with a progressive decrease towards the crypt immature enteroblasts. Cytochrome b and cytochrome oxidase I mt mRNA levels are relatively uniform along the crypt-villus axis, but fractionation experiments showed increased levels in the crypt base. The levels of human mitochondrial transcription factor A are also greater in immature crypt enteroblasts compared with mature villus enterocytes. These results show that the relative levels of mt rRNA and mRNA are distinctly regulated in intestinal epithelial cells according to the crypt-villus position and differentiation status of the cells, and that there are higher mt mRNA and mt TFA levels in the crypts, consistent with increased transcriptional activity during mitochondrial biogenesis in the immature enteroblasts.

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Human small intestinal epithelial cells constitutively express the key elements for antigen processing and the production of exosomes

Blood Cells Molecules and Diseases ◽

10.1016/j.bcmd.2005.05.011 ◽

2005 ◽

Vol 35 (2) ◽

pp. 122-128 ◽

Cited By ~ 35

Author(s):

X.P. Lin ◽

N. Almqvist ◽

E. Telemo

Keyword(s):

Epithelial Cells ◽

Antigen Processing ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Small Intestinal

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Biosynthesis of Microvillus Membrane-Associated Glycoproteins of Small Intestinal Epithelial Cells in Germ-Free and Conventionalized Mice

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a133943 ◽

1982 ◽

Vol 92 (2) ◽

pp. 373-379 ◽

Cited By ~ 12

Author(s):

Yoshinori UMESAKI ◽

Kiyoshi TOHYAMA ◽

Masahiko MUTAI

Keyword(s):

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Germ Free ◽

Microvillus Membrane ◽

Small Intestinal

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Dual Recognition of Sialic Acid and αGal Epitopes by the VP8* Domains of the Bovine Rotavirus G6P[5] WC3 and of Its Mono-reassortant G4P[5] RotaTeq Vaccine Strains

Journal of Virology ◽

10.1128/jvi.00941-19 ◽

2019 ◽

Vol 93 (18) ◽

Cited By ~ 2

Author(s):

Mia Madel Alfajaro ◽

Ji-Yun Kim ◽

Laure Barbé ◽

Eun-Hyo Cho ◽

Jun-Gyu Park ◽

...

Keyword(s):

Epithelial Cells ◽

Blood Group ◽

Intestinal Epithelial Cells ◽

Sialic Acids ◽

Blood Group Antigens ◽

Intestinal Epithelial ◽

Content Type ◽

Group A Rotaviruses ◽

Group A ◽

Small Intestinal

ABSTRACTGroup A rotaviruses, an important cause of severe diarrhea in children and young animals, initiate infection via interactions of the VP8* domain of the VP4 spike protein with cell surface sialic acids (SAs) or histo-blood group antigens (HBGAs). Although the bovine G6P[5] WC3 strain is an important animal pathogen and is also used in the bovine-human reassortant RotaTeq vaccine, the receptor(s) for the VP8* domain of WC3 and its reassortant strains have not yet been identified. In the present study, HBGA- and saliva-binding assays showed that both G6P[5] WC3 and mono-reassortant G4P[5] strains recognized the αGal HBGA. The infectivity of both P[5]-bearing strains was significantly reduced in αGal-free MA-104 cells by pretreatment with a broadly specific neuraminidase or by coincubation with the α2,6-linked SA-specificSambucus nigralectin, but not by the α2,3-linked specific sialidase or byMaackia amurensislectin. Free NeuAc and the αGal trisaccharide also prevented the infectivity of both strains. This indicated that both P[5]-bearing strains utilize α2,6-linked SA as a ligand on MA104 cells. However, the two strains replicated in differentiated bovine small intestinal enteroids and in their human counterparts that lack α2,6-linked SA or αGal HBGA, suggesting that additional or alternative receptors such as integrins, hsp70, and tight-junction proteins bound directly to the VP5* domain can be used by the P[5]-bearing strains to initiate the infection of human cells. In addition, these data also suggested that P[5]-bearing strains have potential for cross-species transmission.IMPORTANCEGroup A rotaviruses initiate infection through the binding of the VP8* domain of the VP4 protein to sialic acids (SAs) or histo-blood group antigens (HBGAs). Although the bovine G6P[5] WC3 strain is an important animal pathogen and is used as the backbone in the bovine-human reassortant RotaTeq vaccine, the receptor(s) for their P[5] VP8* domain has remained elusive. Using a variety of approaches, we demonstrated that the WC3 and bovine-human mono-reassortant G4P[5] vaccine strains recognize both α2,6-linked SA and αGal HBGA as ligands. Neither ligand is expressed on human small intestinal epithelial cells, explaining the absence of natural human infection by P[5]-bearing strains. However, we observed that the P[5]-bearing WC3 and G4P[5] RotaTeq vaccine strains could still infect human intestinal epithelial cells. Thus, the four P[5] RotaTeq vaccine strains potentially binding to additional alternative receptors may be efficient and effective in providing protection against severe rotavirus disease in human.

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