Dde chlorinated insecticide 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (p,p'-DDD) increases intracellular calcium in rat myometrial smooth muscle cells.
A previous report from this laboratory showed that two DDT isomers, o,p'-DDT and p,p'-DDD, increased the frequency of spontaneous oscillatory contractions to a similar extent in isolated rat uterine segments. Because regulation of intracellular calcium is fundamental for the development of oscillatory contractions, the present study examined the effects of p,p'-DDD on intracellular free calcium concentration ([Ca2+]i) in individual rat myometrial smooth muscle cells loaded with the fluorescent Ca2+ indicator fura 2. In the presence of extracellular calcium, 50 and 100 microM p,p'-DDD significantly increased peak [Ca2+]i 586 and 921%, respectively, over basal [Ca2+]i. No significant effect was observed with 10 microM p,p'-DDD. In the absence of extracellular calcium, the response to 100 microM p,p'-DDD was significantly attenuated, with cells averaging a 108% increase in peak [Ca2+]i over basal levels, presumably through Ca2+ release from intracellular stores. Nifedipine and cadmium chloride, blockers of voltage-dependent calcium channels, inhibited 100 microM p,p'-DDD-stimulated increases in [Ca2+]i such that peak [Ca2+]i was increased 250% and 259%, respectively. Because of the prominent inhibition observed with the voltage-dependent calcium channel blockers, the effect of p,p'-DDD on membrane depolarization was examined using a cationic fluorescent indicator of membrane potential, [diS-C2(5)]. A concentration of 50 microM p,p'-DDD depolarized the cells by 35% of maximum during treatment with p,p'-DDD. The data demonstrate that p,p'-DDD increased [Ca2+]i in rat myometrial smooth muscle cells in a concentration-related manner, and that this increase was largely dependent on influx of extracellular calcium through dihydropyridine-sensitive, voltage-dependent calcium channels. The data further show that p,p'-DDD depolarized the plasma membrane, providing a possible mechanism for activation of voltage-dependent calcium channels. Additionally, another calcium source, perhaps an intracellular pool, contributes significantly less to the rise of [Ca2+]i. Whether p,p'-DDD initiates the calcium response by direct actions on the plasma membrane or by other means remains to be determined.
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003632:Dichlorodiphenyldichloroethane / Q000633:toxicity*; D005260:Female; D007306:Insecticides / Q000633:toxicity; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D009215:Myometrium / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014590:Uterine Contraction / Q000187:drug effects
D015220:Calcium Channels; D007306:Insecticides; D002118:Calcium; D003632:Dichlorodiphenyldichloroethane