| Continuous fluorometric measurement of intracellular pH and Ca2+ in perfused salivary gland and pancreas. | |
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MedLine Citation:
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PMID: 8146028 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Intracellular pH (pHi) has been measured in intact, perfused rat mandibular salivary glands loaded with the fluorescent pH indicator BCECF [2',7'-bis(2-carboxyethyl)-5(6)- carboxyfluorescein]. Glands mounted in the cuvette of a conventional bench-top spectro-fluorometer were perfused for 5 min with the acetoxymethyl ester of BCECF and fluorescence was measured ratiometrically at 6-s intervals. The mean value of pHi in glands perfused with a HCO3(-)-free, N-2-hydroxyethylpiperazine-N'-2- ethanesulphonic acid (HEPES)-buffered solution at 37 degrees C was 7.36 +/- 0.01 (n = 52) which is comparable with values obtained by 31P nuclear magnetic resonance (NMR) spectroscopy. NMR data confirmed that the BCECF loading period was accompanied by a transient acidification of the cells, but there was no significant change in the content of the major phosphorus metabolites. Changes in pHi in response to NH4Cl pulses and acetylcholine stimulation were comparable with results reported previously for isolated acini. Additional, preliminary experiments show that the method can also be used to monitor intracellular Ca2+ (using fura-2) in perfused salivary glands, and can be adapted for studies of the isolated, perfused pancreas. |
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Authors:
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J T Seo; M C Steward; J B Larcombe-McDouall; L J Cook; R M Case |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Pflügers Archiv : European journal of physiology Volume: 426 ISSN: 0031-6768 ISO Abbreviation: Pflugers Arch. Publication Date: 1994 Jan |
Date Detail:
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Created Date: 1994-05-03 Completed Date: 1994-05-03 Revised Date: 2009-09-29 |
Medline Journal Info:
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Nlm Unique ID: 0154720 Medline TA: Pflugers Arch Country: GERMANY |
Other Details:
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Languages: eng Pagination: 75-82 Citation Subset: IM |
Affiliation:
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School of Biological Sciences, University of Manchester, UK. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acetylcholine
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pharmacology Amiloride / pharmacology Ammonium Chloride / pharmacology Animals Biological Transport, Active Calcium / metabolism* Fluoresceins / pharmacokinetics Hydrogen-Ion Concentration Intracellular Fluid / metabolism Magnetic Resonance Spectroscopy Male Pancreas / drug effects, metabolism* Perfusion Rats Rats, Sprague-Dawley Sodium-Hydrogen Antiporter / metabolism Spectrometry, Fluorescence Submandibular Gland / drug effects, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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//Wellcome Trust |
| Chemical | |
Reg. No./Substance:
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0/Fluoresceins; 0/Sodium-Hydrogen Antiporter; 12125-02-9/Ammonium Chloride; 2609-46-3/Amiloride; 51-84-3/Acetylcholine; 7440-70-2/Calcium; 85138-49-4/2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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