Isolation and characterization of enterocytes along the intestinal tract of the gilthead seabream ( Sparus aurata L.)

Isolation and characterization of enterocytes along the intestinal tract of the gilthead seabream... Epithelial cells were successfully isolated along the intestine of the gilthead seabream using a dissociation method based on intracellular-like solutions. Biochemical and physiological tests revealed highly viable cells from all intestinal segments. Image analysis was used to identify cell types in the epithelial preparations which were highly enriched in enterocytes (>95%) over mucous cells. Several digestive hydrolases were determined in the isolated cells. Maltase (M), sucrase (S), leucine aminopeptidase (LA), 5′nucleotidase (5′N), but not γ-glutamyl transferase (γ-GT) or alkaline phosphatase (AP) activities were found to be enriched in the epithelial preparations versus the corresponding intestinal homogenates. Comparison of digestive hydrolases revealed the existence of a clear heterogeneity in their expression pattern in the enterocytes, along the intestine. Na + –K + -ATPase, Na + -ATPase and Cl − -ATPase activities were also determined in the membrane fraction of isolated cells. Analyses of enzymatic profiles revealed a clear asymmetry in the distribution of all Mg 2+ -dependent ATPases; that is, maximal Na + –K + - and Na + -ATPase activities were observed in the enterocytes from pyloric caeca, while Cl − -ATPase activity was about twice as high in the enterocytes from anterior and posterior intestines compared with pyloric caeca. This is the first report demonstrating the existence of heterogeneous metabolic and enzymatic profiles in different enterocyte populations from euryhaline teleosts. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Comparative Biochemistry and Physiology - Part A: Molecular & Integrative Physiology Elsevier

Isolation and characterization of enterocytes along the intestinal tract of the gilthead seabream ( Sparus aurata L.)

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Publisher
Elsevier
Copyright
Copyright © 2004 Elsevier Inc.
ISSN
1095-6433
eISSN
1531-4332
D.O.I.
10.1016/j.cbpb.2004.06.013
Publisher site
See Article on Publisher Site

Abstract

Epithelial cells were successfully isolated along the intestine of the gilthead seabream using a dissociation method based on intracellular-like solutions. Biochemical and physiological tests revealed highly viable cells from all intestinal segments. Image analysis was used to identify cell types in the epithelial preparations which were highly enriched in enterocytes (>95%) over mucous cells. Several digestive hydrolases were determined in the isolated cells. Maltase (M), sucrase (S), leucine aminopeptidase (LA), 5′nucleotidase (5′N), but not γ-glutamyl transferase (γ-GT) or alkaline phosphatase (AP) activities were found to be enriched in the epithelial preparations versus the corresponding intestinal homogenates. Comparison of digestive hydrolases revealed the existence of a clear heterogeneity in their expression pattern in the enterocytes, along the intestine. Na + –K + -ATPase, Na + -ATPase and Cl − -ATPase activities were also determined in the membrane fraction of isolated cells. Analyses of enzymatic profiles revealed a clear asymmetry in the distribution of all Mg 2+ -dependent ATPases; that is, maximal Na + –K + - and Na + -ATPase activities were observed in the enterocytes from pyloric caeca, while Cl − -ATPase activity was about twice as high in the enterocytes from anterior and posterior intestines compared with pyloric caeca. This is the first report demonstrating the existence of heterogeneous metabolic and enzymatic profiles in different enterocyte populations from euryhaline teleosts.

Journal

Comparative Biochemistry and Physiology - Part A: Molecular & Integrative PhysiologyElsevier

Published: Sep 1, 2004

References

  • A rabbit jejunal isolated enterocyte preparation suitable for transport studies
    Brown, P.D.; Sepulveda, F.V.
  • Dietary lipid level affects fatty acid composition and hydrolase activities of intestinal brush border membrane in seabass
    Cahu, C.L.; Zambonino Infante, J.L.; Corraze, G.; Coves, D.
  • Digestive enzymes in 11 freshwater teleost fish species in relation to food habit and niche segregation
    Chakrabarti, I.; Gani, Md.A.; Chaki, K.K.; Sur, R.; Misra, K.K.
  • Regulation of brush-border enzyme activities and enterocyte migration rates in mouse small intestine
    Ferraris, R.P.; Villenas, S.A.; Diamond, J.
  • Activity of intestinal mucosal brush border membrane enzymes in relation to the aquaculture fish species
    Harpaz, S.; Uni, Z.
  • Brush-border amino acid transport mechanisms in carnivorous eel intestine
    Storelli, C.; Sebastiano Vilella, M.; Romano, P.; Maffia, M.; Cassano, G.

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