Effect of lead on cytoskeletal proteins expressed in E14 mesencephalic primary cultures.
Several lines of evidence indicated that Pb exposure in vivo and in vitro altered neurite morphology in central and peripheral neurons. The present report shows that neurite length in mesencephalic primary cultures, consisting of neurons and glia, was decreased by Pb exposure when serum factors, presumably essential for glial functions, were absent in the culture medium. We studied whether a serum factor might control the mechanisms involved in the uptake and accumulation of Pb and its effect on cytoskeleton proteins. The total amount of Pb taken up in cell cultures was measured by atomic absorption spectroscopy and appeared to be down-regulated by a non-albumin-like serum component. In presence of serum, Pb exposure failed to alter cytoskeletal proteins. Instead, in serum-free neurobasal medium, Pb uptake failed to reach saturation within 6 h. Western blot analysis showed that the tau, 280 kDa MAP-2b, 70 kDa MAP-2c and GAP-43 protein bands were decreased 24 h after a 3 h exposure to 3 or 6 microM Pb in absence of serum. However, if cultures were maintained in serum-containing media after a 3 h Pb exposure without serum, the immunoblots did not differ from those of controls. It can be inferred that a serum factor prevents cytoskeletal protein alterations by Pb. In serum free medium, Pb that is primarily scavenged by the metallothionein I/II isoforms present in glial cells, may bind to thiol residues of proteins involved in either oxidative stress response or transcriptional regulation of cytoskeletal proteins.
Scortegagna
M
M
Laboratory of Molecular Immunology, NHLBI, Bethesda, MD 20892-1674, USA.
Chikhale
E
E
Hanbauer
I
I
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005260:Female; D019922:GAP-43 Protein / Q000378:metabolism; D007854:Lead / Q000493:pharmacokinetics / Q000633:toxicity*; D008636:Mesencephalon / Q000187:drug effects* / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013054:Spectrophotometry, Atomic; D014404:Tubulin / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
D003598:Cytoskeletal Proteins; D019922:GAP-43 Protein; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D016875:tau Proteins; D007854:Lead