Perturbation by the PCB mixture aroclor 1254 of GABA(A) receptor-mediated calcium and chloride responses during maturation in vitro of rat neocortical cells.
GABA(A) receptors are targets of highly chlorinated environmental chemicals and have important roles in developing neurons. As such, we examined effects of polychlorinated biphenyls (PCBs) on GABA(A) receptor responses in primary cultures of rat neocortical cells using fluorescence imaging techniques. Between days in vitro (DIV) 5 and 8, the effect of GABA(A) receptor stimulation switched from excitatory (Ca(2+) entry following a Cl(-) efflux; DIV </=6) to inhibitory (Cl(-) influx without a Ca(2+) rise; DIV >/=7). GABA(A)-receptor-stimulated increases in [Ca(2+)](i) were diminished in a concentration-dependent (1-20 microM) manner following 1 h of exposure to the PCB mixture Aroclor 1254 (A1254), with significant reductions at concentrations as low as 2 microM. A1254 (1-20 microM) also led to concentration-dependent increases in basal [Ca(2+)](i), irrespective of DIV. A1254 (10 and 20 microM) significantly increased basal Ca(2+)(i); the Ca(2+)(i) was elevated to 426 +/- 39 nM by 20 microM A1254 but this concentration was not cytotoxic at 1 h. In addition, the mixture, A1254, as well as ortho- and non-ortho-chlorinated PCB congeners (IUPAC Nos. 4, 15, 126, and 138; 5-10 microM) individually decreased GABA(A)-stimulated Ca(2+)(i) responses and this tended not to depend on increases in basal Ca(2+)(i). In cultures DIV 7 and older, A1254 (20 microM) also impaired inhibitory GABA(A) responses as evidenced by an approximately 50% reduction of GABA(A)-stimulated Cl(-) influx (from approximately 6 to 8 mM net accumulation in controls). The results demonstrate that: (1) GABA(A) receptor increases in Ca(2+)(i) and Cl(-)(i) are inhibited by 2-20 microM A1254, regardless of whether the responses are at excitatory or inhibitory stages of development; (2) Ca(2+)(i) homeostasis in cortical cells is disrupted by 10 microM A1254; yet (3) disruption of excitatory GABA(A) responses by A1254 or PCB congeners does not necessarily depend on impaired Ca(2+) homeostasis. These novel observations suggest that GABA(A) receptor responses are a sensitive target for PCB effects in the rat developing nervous system.
Inglefield
J R
JR
Neurotoxicology Division, National Health and Environmental Effects Research Laboratory, Research Triangle Park, North Carolina 27711, USA.
Shafer
T J
TJ
D000818:Animals; D013956:Antithyroid Agents / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism*; D020111:Chlorodiphenyl (54% Chlorine) / Q000633:toxicity*; D004347:Drug Interactions; D005456:Fluorescent Dyes; D016257:Fura-2; D018755:GABA Agonists / Q000494:pharmacology; D009118:Muscimol / Q000494:pharmacology; D019579:Neocortex / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D011078:Polychlorinated Biphenyls / Q000633:toxicity*; D051381:Rats; D020318:Rats, Long-Evans; D011963:Receptors, GABA-A / Q000187:drug effects*
D013956:Antithyroid Agents; D002712:Chlorides; D005456:Fluorescent Dyes; D018755:GABA Agonists; D011963:Receptors, GABA-A; D020111:Chlorodiphenyl (54% Chlorine); D009118:Muscimol; D011078:Polychlorinated Biphenyls; D002118:Calcium; D016257:Fura-2