Abstract
To develop techniques for studying transport properties and secretory function of selected cell types in the gastric mucosa, separated fractions of dispersed canine fundic mucosal cells were placed in short-term culture to form epithelial monolayers. Cell fractions enriched in either chief, parietal, or mucous cells were prepared by using counterflow centrifugation and were plated on type I collagen. An epithelial monolayer formed by approximately equal to 36 hr. Immunofluorescence with an antipepsinogen I antibody revealed pepsinogen-containing granules in greater than 95% of the cells, regardless of whether the monolayers were formed from the mucous, chief, or parietal cell-enriched fractions. Upon achieving confluency, chief cell monolayers were mounted in Ussing chambers to study their electrical properties. Under basal conditions, monolayers (n = 6) had a spontaneous potential difference (PD) (+/- SEM) of 26 +/- 4 mV (apical surface negative), a short-circuit current (Isc) (+/- SEM) of 16 +/- 2 microA/cm2, and a transepithelial resistance (R) (+/- SEM) of 1,480 +/- 210 omega X cm2. Histamine increased the short-circuit current, an effect blocked by an H2-receptor antagonist. Seventy percent of the spontaneous PD was amiloride sensitive, suggesting sodium absorption accounted for a major component of the PD. These preparative techniques yield highly enriched chief cell monolayers, which maintain morphological and functional cellular differentiation for greater than 48 hr in culture, thus allowing study of oriented functions of a selected cell type. The present studies indicate that an H2 receptor enhances electrogenic ion transport in chief cell monolayers, indicating that histamine can act on fundic mucosal cells other than just parietal cells.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Berstad A. A modified hemoglobin substrate method for the estimation of pepsin in gastric juice. Scand J Gastroenterol. 1970;5(5):343–348. [PubMed] [Google Scholar]
- Bisbee C. A., Machen T. E., Bern H. A. Mouse mammary epithelial cells on floating collagen gels: transepithelial ion transport and effects of prolactin. Proc Natl Acad Sci U S A. 1979 Jan;76(1):536–540. doi: 10.1073/pnas.76.1.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cereijido M., Robbins E. S., Dolan W. J., Rotunno C. A., Sabatini D. D. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. doi: 10.1083/jcb.77.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Emerman J. T., Pitelka D. R. Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes. In Vitro. 1977 May;13(5):316–328. doi: 10.1007/BF02616178. [DOI] [PubMed] [Google Scholar]
- Guth P. H., Moler T. L., Smith E. H1 and H2 histamine receptors in rat gastric submocosal arterioles. Microvasc Res. 1980 May;19(3):320–328. doi: 10.1016/0026-2862(80)90051-5. [DOI] [PubMed] [Google Scholar]
- Kitahara S., Fox K. R., Hogben C. A. Acid secretion, Na+ absorption, and the origin of the potential difference across isolated mammalian stomachs. Am J Dig Dis. 1969 Apr;14(4):221–238. doi: 10.1007/BF02235951. [DOI] [PubMed] [Google Scholar]
- Logsdon C. D., Bisbee C. A., Rutten M. J., Machen T. E. Fetal rabbit gastric epithelial cells cultured on floating collagen gels. In Vitro. 1982 Mar;18(3 Pt 1):233–242. doi: 10.1007/BF02618576. [DOI] [PubMed] [Google Scholar]
- Misfeldt D. S., Hamamoto S. T., Pitelka D. R. Transepithelial transport in cell culture. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212–1216. doi: 10.1073/pnas.73.4.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Misfeldt D. S., Sanders M. J. Transepithelial transport in cell culture: D-glucose transport by a pig kidney cell line (LLC-PK1). J Membr Biol. 1981 Mar 15;59(1):13–18. doi: 10.1007/BF01870816. [DOI] [PubMed] [Google Scholar]
- Samloff I. M., Liebman W. M. Cellular localization of the group II pepsinogens in human stomach and duodenum by immunofluorescence. Gastroenterology. 1973 Jul;65(1):36–42. [PubMed] [Google Scholar]
- Soll A. H. The actions of secretagogues on oxygen uptake by isolated mammalian parietal cells. J Clin Invest. 1978 Feb;61(2):370–380. doi: 10.1172/JCI108947. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wollin A., Soll A. H., Samloff I. M. Actions of histamine, secretin, and PGE2 on cyclic AMP production by isolated canine fundic mucosal cells. Am J Physiol. 1979 Nov;237(5):E437–E443. doi: 10.1152/ajpendo.1979.237.5.E437. [DOI] [PubMed] [Google Scholar]