Comparative effects of sodium pyrithione evoked intracellular calcium elevation in rodent and primate ventral horn motor neurons.
Oral administration of sodium pyrithione (NaP) causes hindlimb weakness in rodents, but not in primates. Previous work using Aplysia neurons has demonstrated that NaP produces a persistent influx of Ca(2+) ions across the plasma membrane. To determine whether this also occurs in mammalian neurons and whether this could underlie the inter-species difference between rodents and primates, we have tested the effects of NaP on intracellular Ca(2+) levels ([Ca(2+)](i)) in rat and monkey motor neurons in vitro. Motor neurons present in spinal cord slices from rhesus monkey embryos (E37 and 56) and from rat E16 were dissected and cultured on glass coverslips. Following 2 weeks (rhesus) or 2-3 days (rat) in culture, neurons were loaded with fura-PE3/AM, and examined for [Ca(2+)](i) changes in response to NaP. Rhesus motor neurons were identified by immunostaining for Islet-1 (MN specific antigen) and neuron specific enolase (NSE). Motor neurons from both species exhibited dose-dependent NaP-evoked increases in [Ca(2+)](i) However, the dose-response curve for the Rhesus motor neurons was significantly shifted to the right of the rat dose-response curve, whereas the overall amplitude of the Ca(2+) rise was similar in both species. As shown previously for the Aplysia neurons, the action of NaP is attenuated by SKF 96365, an inhibitor of store-operated calcium entry. In contrast the action of NaP is unaffected by nifedipine and tetrodotoxin, blockers of voltage-dependent Ca(2+) and Na(+) channels, respectively, or by ouabain, an inhibitor of the plasma membrane Na(+)/K(+) ATPase. Our results indicate that the NaP-induced increase in [Ca(2+)](i) is conserved across species and suggest that the toxicological sensitivity of rodent over primate to pyrithione could be due to the enhanced sensitivity of rodent motor neurons to NaP-evoked intracellular Ca(2+) elevation.
Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520-8066, USA.
Ronald J
RJ
Knox
Kim L
KL
Keen
Laurelee
L
Luchansky
Ei
E
Terasawa
Hugh
H
Freyer
Steven J
SJ
Barbee
Leonard K
LK
Kaczmarek
D000818:Animals; D000870:Anterior Horn Cells / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D008253:Macaca mulatta; D011725:Pyridines / Q000008:administration & dosage*; D051381:Rats; D013045:Species Specificity; D013871:Thiones / Q000008:administration & dosage*
D011725:Pyridines; D013871:Thiones; C008704:pyrithione; D002118:Calcium