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Determination of ions that are actively transported across porcine vas deferens epithelia
Author(s) -
Devlin Sarah Beth,
Schultz Bruce D
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.759.6
Subject(s) - bumetanide , vas deferens , forskolin , amiloride , chemistry , ussing chamber , ion transporter , secretion , biophysics , endocrinology , sperm , medicine , epithelial polarity , apical membrane , sodium , biology , andrology , in vitro , biochemistry , cell , membrane , organic chemistry
An appropriate fluid environment is required for sperm development, storage and activation, although the underlying mechanisms to account for electrolyte secretion into the male reproductive duct are not fully defined. Radiotracer flux experiments were conducted to test for unidirectional and net Na + , Cl − , and K + transport across epithelial cells that were isolated from porcine vas deferens and grown in culture. Epithelial monolayers were exposed to the corticosteroid hormone dexamethasone for 72 hours before being mounted in modified Ussing flux Chambers for ion transport analysis. The apical and basolateral aspects of the epithelia were separately exposed, in paired monolayers, to tracer amounts of 22 Na + , 36 Cl − , or 86 Rb + . Sampling was conducted over twenty minute periods in four cumulative conditions: basal, amiloride, forskolin, and bumetanide. Samples were analyzed using scintillation fluorimetry. Results indicate that amiloride sensitive Na + absorption is present at the outset of sampling. Subsequently, forskolin was shown to stimulate bumetanide sensitive Cl − secretion across these epithelia. No net 86 Rb + flux was observed, which indicates no net K + movement. Na + and Cl − flux may not fully account for the observed short circuit current. Thus, additional studies are required to test for the contribution of HCO 3 − to stimulated secretion, which may contribute to sperm activation.