Electrophysiological effects of chilotoquine on tight junctions of immature rat Sertoli cells in vitro.
We investigated the effect of CQ, an antimalarial drug with antiprotease activity, and NH4Cl, a related amines on the development of intercellular tight junctions in cultured immature rat Sertoli cells. Sertoli cells were seeded in serum-free defined medium at a density of 3 x 10(6) cells/0.64 cm2/well on Matrigel-covered Millicell-HA filters. CQ (1 microM and 2 microM) or NH4Cl (6.25 mM and 12 mM) was added to the outer (basal) compartment of the bicameral system either on day 1 or day 7 of the culture. Formation of tight junctions was monitored by measurement of the transepithelial resistance (TER) at 24 hr intervals using an impedance meter. TER in untreated controls was 50 omega/cm2 on day 1, increased progressively to 80 omega/cm2 by day 7 and plateaued until day 12. The cells treated from day 1 with CQ showed dose-dependent progressive increase in TER until day 12, reaching 191 omega/cm2 in cells treated with 1 microM concentration. In cells treated with CQ starting from day 7 of culture onwards, TER patterns were similar to those noted following exposure to chloroquine from day 1. Also in cultures containing NH4Cl, in comparison to the control, the increase in TER was significantly higher. These observations demonstrate that CQ and HN4Cl promote tight junction formation between immature rat Sertoli cells invitro suggesting that antiproteases may be involved in the formation of blood-testis barrier.
Okanlawon
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Department of Cell Biology, Georgetown University Medical Center, Washington DC, 20007, USA.
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D000643:Ammonium Chloride / Q000009:adverse effects*; D000818:Animals; D000831:Animals, Newborn; D000962:Antimalarials / Q000009:adverse effects*; D001814:Blood-Testis Barrier / Q000187:drug effects; D002478:Cells, Cultured; D002738:Chloroquine / Q000009:adverse effects*; D015194:Diffusion Chambers, Culture; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D017097:Electric Impedance; D004594:Electrophysiology; D010450:Endopeptidases / Q000187:drug effects; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D019108:Tight Junctions / Q000187:drug effects*
D000962:Antimalarials; D000643:Ammonium Chloride; D002738:Chloroquine; D010450:Endopeptidases