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Tyrosine Hydroxylase mRNA Expression by Dopaminergic Neurons in Culture: Effect of 1 ‐Methyl‐4‐Phenylpyridinium Treatment
Author(s) -
Beck Klaus D.,
Knusel Beat,
Pasinetti Giulio,
Michel Patrick P.,
Zawadzka Halina,
Goldstein Menek,
Hefti Franz
Publication year - 1991
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1991.tb03782.x
Subject(s) - tyrosine hydroxylase , dopaminergic , tyrosine 3 monooxygenase , chemistry , medicine , endocrinology , messenger rna , tyrosine , dopamine , biology , biochemistry , gene
To enable us to study expression of tyrosine hydroxylase [TH; tyrosine 3‐monooxygenase; L‐tyrosine tetrahydropteridine: oxygen oxidoreductase (3‐hydroxylating); EC 1.14.16.2] as a measure of dopaminergic neuron function in future experiments, methods were developed to quantify TH mRNA levels in cultures of dopaminergic mesencephalic cells. The model of selective dopaminergic toxicity of 1‐methyl‐4‐phenylpyridinium (MPP + ) was used to verify the specificity of our methods. Fetal (embryonic day 15) rat ventral mesencephalic cell cultures were treated with 15 μM MPP + for 48 h, conditions previously shown to reduce the number of TH‐immunoreactive neurons, TH activity, and dopamine uptake to 5–10% of control values. This treatment decreased the number of neurons labeled by TH in situ hybridization to 9% of untreated controls and caused a strong reduction of the abundance of TH mRNA in Northern blots. Our findings establish TH mRNA expression as a parameter for future studies of toxic and trophic effects on cultured dopaminergic neurons, and they support the view that MPP + destroys dopaminergic neurons.