Stage-dependent vulnerability of fetal mesencephalic neuroprogenitors towards dopaminergic neurotoxins.
Although extensive knowledge exists on selective vulnerability of dopaminergic neurons against parkinsonism-inducing neurotoxins, there is a complete lack of such data on immature neuroprogenitors. Here we investigated the toxicity of 1-methyl-4-phenylpyridinium (MPP+), 6-hydroxydopamine (6-OHDA) and the free radical generator H2O2 on various developmental stages of predopaminergic mesencephalic neuroprogenitors (mNPCs) to evaluate stage-dependency of selective dopaminergic neurotoxicity. Striatal NPCs (sNPCs) without dopaminergic differentiation potential served as controls. Exposure of both undifferentiated NPCs to MPP+ resulted in concentration-dependent cell death at concentrations of >10 microM after 72 h without differences between both cell types, while 6-OHDA led to relevant cell death at 1000 microM after 24h with significant higher sensitivity of mNPCs compared to sNPCs. H2O2 did not induce relevant cell death in all cell types. In NPC cultures differentiated for 14 days, MPP+ showed enhanced toxicity compared to the undifferentiated counterparts, but no significant differences between both NPC type and differentiation conditions. 6-OHDA showed similar toxicity pattern in differentiated compared to undifferentiated NPCs. By evaluating the toxicity of MPP+ on MAP2ab+ neurons derived from both mNPCs and sNPCs as well as tyrosine hydroxylase (TH)+ dopaminergic cells from mNPCs, we found concentration-dependent cell death of all cell types with no increased vulnerability of TH+ cells. Primary TH+ neurons showed significantly higher vulnerability to MPP+. Together, we demonstrated stage-dependent vulnerability of NPCs towards dopaminergic neurotoxins, but no selective vulnerability of NPC-derived TH+ dopaminergic cells towards MPP+. This cell system seems not suitable as a screening tool for selective dopaminergic toxicity.
Department of Neurology, University of Ulm, Ulm, Germany.
Michael
M
Sabolek
Ines
I
Mieskes
Thomas
T
Lenk
Vera
V
Lehmensiek
Andreas
A
Hermann
Johannes
J
Schwarz
Alexander
A
Storch
D015655:1-Methyl-4-phenylpyridinium / Q000494:pharmacology; D000818:Animals; D002452:Cell Count / Q000379:methods; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000378:metabolism; D008636:Mesencephalon / Q000166:cytology* / Q000196:embryology*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D016627:Oxidopamine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
D016207:Cytokines; D051100:Glial Cell Line-Derived Neurotrophic Factor; C489787:MAP2 protein, rat; D008869:Microtubule-Associated Proteins; D005576:Colforsin; D016627:Oxidopamine; D006861:Hydrogen Peroxide; D015655:1-Methyl-4-phenylpyridinium; D004298:Dopamine