Possible molecular targets of halogenated aromatic hydrocarbons in neuronal cells.
Halogenated aromatic hydrocarbon including polychlorinated biphenyls (PCBs) are persistent and bioaccumulative environmental toxicants. Although health effects associated with exposure to these chemicals, including motor dysfunction and impairment in memory and learning, have been identified, their molecular site of action is unknown. Previous study from this laboratory demonstrated that, while ortho PCBs perturbed intracellular signaling mechanisms including Ca2+ homeostasis, receptor-mediated inositol phosphate production and translocation of PKC, non-ortho PCBs did not. Since PKC signaling pathway is implicated in the modulation of motor behavior, as well as learning and memory, and the roles of PKC are isoform-specific, we have now studied the effects of two structurally distinct PCBs on isoforms of PKC in cerebellar granule cell culture model. Cells were exposed to 2,2'-dichlorobiphenyl (ortho PCB; 2,2'-DCB) or 4,4'-dichlorobiphenyl (non-ortho PCB; 4,4'-DCB) for 15 min, respectively, and subsequently fractionated and immunoblotted against the selected PKC monoclonal antibodies (alpha, gamma, delta, epsilon, lambda, iota). While 2,2'-DCB induced a translocation of PKC-alpha [cytosol (% control): 54 +/- 12 at 25 microM and 66 +/- 10 at 50 microM; membrane (% control): 186 +/- 37 at 25 microM and 200 +/- 48 at 50 microM] and -epsilon [cytosol (% control): 92 +/- 12 at 25 microM and 97 +/- 15 at 50 microM; membrane (% control): 143 +/- 23 at 25 microM and 192 +/- 24 at 50 microM] from cytosol to membrane fraction in a concentration-dependent manner, 4,4'-DCB had no effects. 2,2'-DCB induced translocation of PKC-alpha was blocked by pretreatment with sphingosine, suggesting a possible role of sphingolipid pathway. Although reports on implication of PKC-gamma with learning and memory are relatively extensive, the expression of this particular isoform in the primary cerebellar granule cells was below the detectable level. PKC-delta, -lambda and -iota were present in these cells, but were not altered by PCB exposure. These results suggest that the effects of 2,2'-DCB on PKC is isoform-dependent and PKC-alpha as well as PKC-epsilon may be target molecules for ortho-PCBs in neuronal cells.
Yang
J H
JH
Neurotoxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA.
Kodavanti
P R
PR
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D007527:Isoenzymes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D008566:Membranes / Q000187:drug effects / Q000201:enzymology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D011078:Polychlorinated Biphenyls / Q000494:pharmacology*; D011493:Protein Kinase C / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D020318:Rats, Long-Evans; D013110:Sphingosine / Q000494:pharmacology; D013329:Structure-Activity Relationship
D007527:Isoenzymes; C012280:2,2'-dichlorobiphenyl; D011078:Polychlorinated Biphenyls; C029861:4,4'-dichlorobiphenyl; D011493:Protein Kinase C; D013110:Sphingosine