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Regulated anion secretion in cultured epithelia from Sertoli cells of immature rats
Author(s) -
Ko W. H.,
Chan H. C.,
Chew S. B.,
Wong P. Y. D.
Publication year - 1998
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1111/j.1469-7793.1998.471be.x
Subject(s) - dids , forskolin , transepithelial potential difference , thapsigargin , endocrinology , medicine , ion transporter , adenosine , extracellular , chemistry , membrane potential , reversal potential , biology , biophysics , stimulation , patch clamp , biochemistry , electrophysiology , membrane
1 Cultured epithelia of Sertoli cells from prepubertal rats were grown on Matrigel‐coated millipore filters for short‐circuit current ( I sc ) measurements. Under basal conditions, these epithelia exhibited a ‘zero’ transepithelial potential difference, a ‘zero’ short‐circuit current and a transepithelial resistance of 60 Ω cm 2 . 2 Forskolin (100 μ m ) and 8‐(4‐chlorophenylthio)‐cAMP (cpt‐cAMP) (100 μ m ) added to the apical side stimulated the I sc (forskolin, peak Δ I sc = 1.32 ± 0.16 μA cm −1 ; cpt‐cAMP, peak Δ I sc = 0.88 ± 0.16 μA cm −2 ). 3 ATP (100 μ m ) added apically elicited a I sc response (peak Δ I sc = 6.45 ± 0.28 μA cm −2 ) which was similar in magnitude to that of 1 μ m thapsigargin (peak Δ I sc = 6.09 ± 0.44 μA cm −2 ). The potency of the responses to other nucleotides: UTP ≥ ATP > ADP >> AMP = adenosine indicates the involvement of a mixture of P2Y receptors. 4 Removal of extracellular Cl − and HCO 3 − reduced the I sc response to ATP by 70 % and 40 %, respectively. Removal of K + had no effect, whereas removal of Na + attenuated the I sc response. 5 The response to ATP was insensitive to agents known to block anion secretion (except apical diphenylamine‐2‐carboxylate (DPC) and DIDS). The resistance to perturbation by pharmacological agents may be a unique property of the seminiferous epithelium. 6 Whole‐cell current recordings in cultured rat Sertoli cells demonstrated a DIDS‐sensitive outwardly rectifying Cl − conductance with activating and inactivating characteristics at depolarizing and hyperpolarizing voltages, respectively. 7 The stimulation of electrogenic ion transport by ATP may be part of a complex mechanism regulating fluid secretion by the testis. Cultured Sertoli cell epithelia are shown to provide a useful model to investigate transepithelial transport in the seminiferous epithelium.