Premium
Characterization of organotypic ventral mesencephalic cultures from embryonic mice and protection against MPP + toxicity by GDNF
Author(s) -
Jakobsen B.,
Gramsbergen J. B.,
Møller Dall A.,
Rosenblad C.,
Zimmer J.
Publication year - 2005
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1111/j.1460-9568.2005.04138.x
Subject(s) - glial cell line derived neurotrophic factor , dopaminergic , dopamine , tyrosine hydroxylase , neurotrophic factors , neuron , dopamine transporter , medicine , endocrinology , neuroprotection , biology , chemistry , pharmacology , neuroscience , receptor
We characterized organotypic ventral mesencephalic (VM) cultures derived from embryonic day 12 (E12) mice (CBL57/bL6) in terms of number of dopaminergic neurons, cell soma size and dopamine production in relation to time in vitro and tested the effects of 1‐methyl‐4‐phenylpyridinium (MPP + ) and glial derived neurotrophic factor (GDNF) to validate this novel culture model. Dopamine production and dopaminergic neuron soma size increased dramatically with time in vitro , whereas the number of dopamine neurons declined by approximately 30% between week 1 and week 2, which was further reduced after week 4. GDNF treatment (100 ng/mL) increased dopaminergic neuron soma size (up to 43%) and DOPAC production (approximately three‐fold), but not the number of dopamine neurons in control cultures. One‐week‐old cultures were more vulnerable to MPP + , than three‐week‐old cultures. The EC 50 for dopamine depletion after 2 days exposure and 15 days of recovery were 0.6 and 7 µm, respectively. Both pre‐treatment and post‐treatment with GDNF are important to obtain maximal protection against MPP + toxicity. In one‐week‐old cultures (5 µm MPP + , 2 days) GDNF provided potent neuroprotection with dopamine contents reaching control levels and number of tyrosine hydroxylase (TH) + cells up to 80% of control, but in three‐week‐old cultures (10 µm MPP + , 2 days) the protective potential of GDNF was markedly reduced. Long recovery periods after MPP + exposure are required to distinguish between reversible or irreversible toxic and/or trophic effects.