Journal article
Functional characterization of Na(+)/H(+) exchangers in primary cultures of prairie dog gallbladder
The Journal of membrane biology, v 197(2)
15 Jan 2004
PMID: 15014914
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Gallbladder Na(+) absorption is linked to gallstone formation in prairie dogs. We previously reported Na(+)/H(+) exchanger (NHE1-3) expression in native gallbladder tissues. Here we report the functional characterization of NHE1, NHE2 and NHE3 in primary cultures of prairie dog gallbladder epithelial cells (GBECs). Immunohistochemical studies showed that GBECs grown to confluency are homogeneous epithelial cells of gastrointestinal origin. Electron microscopic analysis of GBECs demonstrated that the cells form polarized monolayers characterized by tight junctions and apical microvilli. GBECs grown on Snapwells exhibited polarity and developed transepithelial short-circuit current, I(sc), (11.6 +/- 0.5 microA. cm(-2)), potential differences, V(t) (2.1 +/- 0.2 mV), and resistance, R(t) (169 +/- 12 omega. cm(2)). NHE activity in GBECs assessed by measuring dimethylamiloride-inhibitable (22)Na(+) uptake under a H(+) gradient was the same whether grown on permeable Snapwells or plastic wells. The basal rate of (22)Na(+) uptake was 21.4 +/- 1.3 nmol x mg prot(-1) x min(-1), of which 9.5 +/- 0.7 (approximately 45%) was mediated through apically-restricted NHE. Selective inhibition with HOE-694 revealed that NHE1, NHE2 and NHE3 accounted for approximately 6%, approximately 66% and approximately 28% of GBECs' total NHE activity, respectively. GBECs exhibited saturable NHE kinetics ( V(max) 9.2 +/- 0.3 nmol x mg prot(-1) x min(-1); K(m) 11.4 +/- 1.4 m M Na(+)). Expression of NHE1, NHE2 and NHE3 mRNAs was confirmed by RT-PCR analysis. These results demonstrate that the primary cultures of GBECs exhibit Na(+) transport characteristics similar to native gallbladder tissues, suggesting that these cells can be used as a tool for studying the mechanisms of gallbladder ion transport both under physiologic conditions and during gallstone formation.
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Details
- Title
- Functional characterization of Na(+)/H(+) exchangers in primary cultures of prairie dog gallbladder
- Creators
- S C Narins - Drexel UniversityE H Park - Drexel UniversityR Ramakrishnan - Drexel UniversityF U Garcia - Drexel UniversityJ N Diven - Drexel UniversityB J Balin - Philadelphia College of Osteopathic MedicineC J Hammond - Philadelphia College of Osteopathic MedicineB R Sodam - Drexel UniversityP R Smith - University of Alabama at BirminghamM Z Abedin - Drexel University
- Publication Details
- The Journal of membrane biology, v 197(2)
- Publisher
- Springer Nature
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Pathology (and Laboratory Medicine)
- Web of Science ID
- WOS:000189304100005
- Scopus ID
- 2-s2.0-10744225025
- Other Identifier
- 991019168781204721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology
- Cell Biology
- Physiology