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

Dopido, R; Rodrı́guez, Covadonga; Gómez, Tomás; Acosta, N.G.; Dı́az, Mario

doi:10.1016/j.cbpb.2004.06.013

Cited by 11 publications

(12 citation statements)

References 25 publications

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“…Cell oxygen consumption was neither affected by diet showing values that were in the range previously reported for gilthead seabream enterocytes (Dópido et al, 2004).…”

Section: Discussionsupporting

confidence: 83%

“…After 48 h of starvation, the foregut, including pyloric caeca, was rapidly removed from the carcass, cleaned of adhering adipose tissue, and contents removed with cold (4 ºC) physiological saline solution III (Dópido et al, 2004). The intestine was then filled, immersed and incubated with a hyperosmolar solution.…”

Section: Preparation Of Isolated Enterocytes and Hepatocytesmentioning

confidence: 99%

“…Oxygen consumption was measured on freshly isolated cells in an O 2 -saturated closed system using a Clark electrode connected to a computerized data acquisition system, and processed with (Dópido et al, 2004).…”

Section: Enterocyte and Hepatocyte Viabilitymentioning

confidence: 99%

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Effects of dietary fish oil substitution by Echium oil on enterocyte and hepatocyte lipid metabolism of gilthead seabream (Sparus aurata L.)

Díaz-López

Pérez

Acosta

et al. 2010

Comparative Biochemistry and Physiology Part B: Biochemistry an

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The fatty acid profile of vegetable oils (VOs), together with the poor ability of marine fish to convert polyunsaturated fatty acids (PUFA) to highly unsaturated fatty acids (HUFA), lead to important changes in the nutritional value of farmed fish fed VO, which include increased fat and 18:2n-6 and reduced n-3 HUFA. Echium oil (EO) has a good n-3/n-6 balance as well as an interesting profile with its high content of unusual fatty acids (SDA, was also unaffected by EO in terms of total uptake, incorporation, β-oxidation and elongationdesaturation activities.

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“…Cell oxygen consumption was neither affected by diet showing values that were in the range previously reported for gilthead seabream enterocytes (Dópido et al, 2004).…”

Section: Discussionsupporting

confidence: 83%

Section: Preparation Of Isolated Enterocytes and Hepatocytesmentioning

confidence: 99%

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Effects of dietary fish oil substitution by Echium oil on enterocyte and hepatocyte lipid metabolism of gilthead seabream (Sparus aurata L.)

Díaz-López

Pérez

Acosta

et al. 2010

Comparative Biochemistry and Physiology Part B: Biochemistry an

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“…Cl -uptake is unaffected by mucosal exposure to the anion exchanger inhibitors SITS (4-acetamino-4′-isothiocyanostilbene-2,2′-disulfonic acid) and DIDS (4,4′-diisothiocyanostilbene-2,2′-disulfonic acid) , suggesting that transport may not occur by apical Cl -,HCO 3 -exchange. Interestingly, CFTR appears to localize to the basolateral surface of pavement cells in killifish opercular epithelium in freshwater, while it is not expressed in this cell type in seawater Dópido et al, 2004) in the opercular epithelium of killifish in freshwater.…”

Section: Mechanisms Of Ion Absorptionmentioning

confidence: 97%

Gene expression after freshwater transfer in gills and opercular epithelia of killifish: insight into divergent mechanisms of ion transport

Scott

Claiborne

Edwards

et al. 2005

Journal of Experimental Biology

123

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SUMMARY We have explored the molecular basis for differences in physiological function between the gills and opercular epithelium of the euryhaline killifish Fundulus heteroclitus. These tissues are functionally similar in seawater, but in freshwater the gills actively absorb Na+ but not Cl–, whereas the opercular epithelium actively absorbs Cl– but not Na+. These differences in freshwater physiology are likely due to differences in absolute levels of gene expression (measured using real-time PCR), as several proteins important for Na+ transport, namely Na+,H+-exchanger 2 (NHE2), carbonic anhydrase 2 (CA2),Na+,HCO3–cotransporter 1, and V-type H+-ATPase, were expressed at 3- to over 30-fold higher absolute levels in the gills. In gills, transfer from 10% seawater to freshwater increased the activity of Na+,K+-ATPase by twofold (from 12 h to 7 days), increased the expression of NHE2 (at 12 h) and CA2 (from 12 h to 7 days), and decreased the expression of NHE3 (from 12 h to 3 days). In opercular epithelium, NHE2 was not expressed; furthermore,Na+,K+-ATPase activity was unchanged after transfer to freshwater, CA2 mRNA levels decreased, and NHE3 levels increased. Consistent with their functional similarities in seawater, killifish gills and opercular epithelium expressed Na+,K+-ATPase α1a,Na+,K+,2Cl–cotransporter 1 (NKCC1),cystic fibrosis transmembrane conductance regulator (CFTR)Cl– channel and the signalling protein 14-3-3a at similar absolute levels. Furthermore, NKCC1 and CFTR were suppressed equally in each tissue after freshwater transfer, and 14-3-3a mRNA increased in both. These results provide insight into the mechanisms of ion transport by killifish gills and opercular epithelia, and demonstrate a potential molecular basis for the differences in physiological function between these two organs.

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“…Moreover, primary enterocytes rarely multiply for more than 10 generations, which restricts the number of experiments per cell lot and consequently the comparative value of the results (Bader et al, 2000;Dimier-Poisson et al, 2004;Dopido et al, 2004;Evans et al, 1994;Macartney et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Preliminary characterization of jejunocyte and colonocyte cell lines isolated by enzymatic digestion from adult and young cattle

Loret

Rusu

Moualij

et al. 2009

Research in Veterinary Science

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a b s t r a c tIn the present study we developed an enzymatic approach (through the use of collagenase and dispase) to isolate bovine intestinal epithelial cells. Using this method, freshly isolated jejunocytes could be distinguished from simultaneously isolated colonocytes, as the jejunocytes specifically exhibited the small intestinal peptidase gene transcript, as well as an active alkaline phosphatase. The transformation of both types of cell suspension was performed by retroviral infection, using reproduction-defective viruses bearing the gene coding for the large T antigen of the leukaemia simian virus (SV40). The success of the transfection was demonstrated by (1) a significant increase in cell passage numbers (52-53 vs. 7 passages for nontransfected cells), (2) the detection of both the large T transcript and the large T antigen in transformed cells. Possible contamination and progressive substitution of bovine primocultures by non-bovine lineages available in the laboratory was excluded, as the transformed cells presented a bovine typical karyotype. Most transfected cells kept an epithelial morphology after transformation. They also maintained the expression of FABP and enterocyte specific enzymes (brush-border associated maltase and IAP). However, levels of specific activity of these enzymes were low, suggesting that cell differentiation is not completely achieved under the applied culture conditions.

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

Cited by 11 publications

References 25 publications

Effects of dietary fish oil substitution by Echium oil on enterocyte and hepatocyte lipid metabolism of gilthead seabream (Sparus aurata L.)

Effects of dietary fish oil substitution by Echium oil on enterocyte and hepatocyte lipid metabolism of gilthead seabream (Sparus aurata L.)

Gene expression after freshwater transfer in gills and opercular epithelia of killifish: insight into divergent mechanisms of ion transport

Preliminary characterization of jejunocyte and colonocyte cell lines isolated by enzymatic digestion from adult and young cattle

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