Glucose transport by epithelia prepared from harvested enterocytes

Yasuhiro Kimura, Marie van der Merwe, Stine B. Bering, Himabindu Penmatsa, Veronica G. Conoley, Per T. Sangild, Anjaparavanda P. Naren, Randal Buddington

Research output: Contribution to journalArticle

Abstract

Transformed and cultured cell lines have significant shortcomings for investigating the characteristics and responses of native villus enterocytes in situ. Interpretations of results from intact tissues are complicated by the presence of underlying tissues and the crypt compartment. We describe a simple, novel, and reproducible method for preparing functional epithelia using differentiated enterocytes harvested from the small intestine upper villus of adult mice and preterm pigs with and without necrotizing enterocolitis. Concentrative, rheogenic glucose uptake was used as an indicator of epithelial function and was demonstrated by cellular accumulation of tracer 14 C d-glucose and Ussing chamber based short-circuit currents. Assessment of the epithelia by light and immunofluorescent microscopy revealed the harvested enterocytes remain differentiated and establish cell–cell connections to form polarized epithelia with distinct apical and basolateral domains. As with intact tissues, the epithelia exhibit glucose induced short-circuit currents that are increased by exposure to adenosine and adenosine 5′-monophosphate (AMP) and decreased by phloridzin to inhibit the apical glucose transporter SGLT-1. Similarly, accumulation of 14 C d-glucose by the epithelia was inhibited by phloridzin, but not phloretin, and was stimulated by pre-exposure to AMP and adenosine, apparently by a microtubule-based mechanism that is disrupted by nocodazole, with the magnitudes of responses to adenosine, forskolin, and health status exceeding those we have measured using intact tissues. Our findings indicate that epithelia prepared from harvested enterocytes provide an alternative approach for comparative studies of the characteristics of nutrient transport by the upper villus epithelium and the responses to different conditions and stimuli.

Original languageEnglish (US)
Pages (from-to)39-49
Number of pages11
JournalCytotechnology
Volume67
Issue number1
DOIs
StatePublished - Jan 1 2013

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Enterocytes
Adenosine
Glucose
Epithelium
Tissue
Phlorhizin
Short circuit currents
Phloretin
Nocodazole
Facilitative Glucose Transport Proteins
Colforsin
Adenosine Monophosphate
Nutrients
Microscopic examination
Cells
Health
Necrotizing Enterocolitis
Transformed Cell Line
Microtubules
Health Status

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

Cite this

Kimura, Y., van der Merwe, M., Bering, S. B., Penmatsa, H., Conoley, V. G., Sangild, P. T., ... Buddington, R. (2013). Glucose transport by epithelia prepared from harvested enterocytes. Cytotechnology, 67(1), 39-49. https://doi.org/10.1007/s10616-013-9656-1

Glucose transport by epithelia prepared from harvested enterocytes. / Kimura, Yasuhiro; van der Merwe, Marie; Bering, Stine B.; Penmatsa, Himabindu; Conoley, Veronica G.; Sangild, Per T.; Naren, Anjaparavanda P.; Buddington, Randal.

In: Cytotechnology, Vol. 67, No. 1, 01.01.2013, p. 39-49.

Research output: Contribution to journalArticle

Kimura, Y, van der Merwe, M, Bering, SB, Penmatsa, H, Conoley, VG, Sangild, PT, Naren, AP & Buddington, R 2013, 'Glucose transport by epithelia prepared from harvested enterocytes', Cytotechnology, vol. 67, no. 1, pp. 39-49. https://doi.org/10.1007/s10616-013-9656-1
Kimura Y, van der Merwe M, Bering SB, Penmatsa H, Conoley VG, Sangild PT et al. Glucose transport by epithelia prepared from harvested enterocytes. Cytotechnology. 2013 Jan 1;67(1):39-49. https://doi.org/10.1007/s10616-013-9656-1
Kimura, Yasuhiro ; van der Merwe, Marie ; Bering, Stine B. ; Penmatsa, Himabindu ; Conoley, Veronica G. ; Sangild, Per T. ; Naren, Anjaparavanda P. ; Buddington, Randal. / Glucose transport by epithelia prepared from harvested enterocytes. In: Cytotechnology. 2013 ; Vol. 67, No. 1. pp. 39-49.
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