TY - JOUR
T1 - Characterization of organotypic ventral mesencephalic cultures from embryonic mice and protection against MPP toxicity by GDNF
AU - Jakobsen, B
AU - Gramsbergen, J B
AU - Møller Dall, A
AU - Rosenblad, C
AU - Zimmer, J
PY - 2005/6/1
Y1 - 2005/6/1
N2 - 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 microm, respectively. Both pre-treatment and post-treatment with GDNF are important to obtain maximal protection against MPP(+) toxicity. In one-week-old cultures (5 microm 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 microm 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.
AB - 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 microm, respectively. Both pre-treatment and post-treatment with GDNF are important to obtain maximal protection against MPP(+) toxicity. In one-week-old cultures (5 microm 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 microm 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.
KW - 1-Methyl-4-phenylpyridinium
KW - 3,4-Dihydroxyphenylacetic Acid
KW - Animals
KW - Cell Differentiation
KW - Cell Proliferation
KW - Cell Size
KW - Cells, Cultured
KW - Disease Models, Animal
KW - Dopamine
KW - Dose-Response Relationship, Drug
KW - Glial Cell Line-Derived Neurotrophic Factor
KW - Mice
KW - Mice, Inbred C57BL
KW - Nerve Degeneration
KW - Nerve Growth Factors
KW - Neurons
KW - Neuroprotective Agents
KW - Neurotoxins
KW - Organ Culture Techniques
KW - Parkinsonian Disorders
KW - Recovery of Function
KW - Substantia Nigra
KW - Time Factors
KW - Tyrosine 3-Monooxygenase
U2 - 10.1111/j.1460-9568.2005.04138.x
DO - 10.1111/j.1460-9568.2005.04138.x
M3 - Journal article
C2 - 15978005
SN - 0953-816X
VL - 21
SP - 2939
EP - 2948
JO - European Journal of Neuroscience
JF - European Journal of Neuroscience
IS - 11
ER -