Effect of GDNF on differentiation of cultured ventral mesencephalic dopaminergic and non-dopaminergic calretinin-expressing neurons

• Published in: Brain research Vol. 1036; no. 1; pp. 163 - 172
• Main Authors: Schaller, Benoît, Andres, Robert H., Huber, Alexander W., Meyer, Morten, Pérez-Bouza, Alberto, Ducray, Angélique D., Seiler, Rolf W., Widmer, Hans R.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 02.03.2005; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Biological and medical sciences; Calbindin; Calbindin 2; Calbindins; Calretinin; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell differentiation, maturation, development, hematopoiesis; Cell morphology; Cell physiology; Cell Shape - drug effects; Cell Shape - physiology; Cells, Cultured; Dopamine - metabolism; Fundamental and applied biological sciences. Psychology; gamma-Aminobutyric Acid - metabolism; Glial Cell Line-Derived Neurotrophic Factor; Immunohistochemistry; Mesencephalon - cytology; Mesencephalon - drug effects; Mesencephalon - embryology; Molecular and cellular biology; Nerve Growth Factors - metabolism; Nerve Growth Factors - pharmacology; Neurites - drug effects; Neurites - metabolism; Neurites - ultrastructure; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; Rats; Rats, Sprague-Dawley; S100 Calcium Binding Protein G - metabolism; Serotonin - metabolism; Substantia Nigra - cytology; Substantia Nigra - drug effects; Substantia Nigra - embryology; Tyrosine 3-Monooxygenase - metabolism; Tyrosine hydroxylase; Ventral mesencephalon; Ventral Tegmental Area - cytology; Ventral Tegmental Area - drug effects; Ventral Tegmental Area - metabolism; Glial cell line-derived neurotrophic factor; Calbindin; Development and regeneration; Calretinin; Tyrosine hydroxylase; Ventral mesencephalon; Cell morphology; Calcium; Rat; Central nervous system; Tyrosine 3-monooxygenase; Binding protein; Glial cell line derived neurotrophic factor; Dopamine; Enzyme; Rodentia; Midbrain; Catecholamine; In vitro; Vertebrata; Mammalia; Morphology; Neurotransmitter; Dopaminergic neuron; Oxidoreductases; Differentiation
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2004.12.054

Glial cell line-derived neurotrophic factor (GDNF) is a potent survival factor for ventral mesencephalic (VM) dopaminergic neurons. Subpopulations of dopaminergic and non-dopaminergic VM neurons express the calcium-binding proteins calbindin (CB) and calretinin (CR). Characterization of the actions of GDNF on distinct subpopulations of VM cells is of great importance for its potential use as a therapeutic molecule and for understanding its role in neuronal development. The present study investigated the effects of GDNF on the survival and morphological differentiation of dopaminergic and non-dopaminergic neurons in primary cultures of embryonic day (E) 18 rat VM. As expected from our results obtained using E14 VM cells, GDNF significantly increased the morphological complexity of E18 CB-immunoreractive (CB-ir), tyrosine hydroxylase (TH)-ir, and CR-ir neurons and also the densities of CB-ir and TH-ir neurons. Interestingly, densities of E18 CR-ir neurons, contrarily to our previous observations on E14 CR-ir neurons, were significantly higher after GDNF treatment (by 1.5-fold). Colocalization analyses demonstrated that GDNF increased the densitiy of dopaminergic neurons expressing CR (TH+/CR+/CB−), while no significant effects were observed for TH−/CR+/CB− cell densities. In contrast, we found that GDNF significantly increased the total fiber length (2-fold), number of primary neurites (1.4-fold), number of branching points (2.5-fold), and the size of neurite field per neuron (1.8-fold) of the non-dopaminergic CR-expressing neurons (TH−/CR+/CB−). These cells were identified as GABA-expressing neurons. In conclusion, our findings recognize GDNF as a potent differentiation factor for the development of VM dopaminergic and non-dopaminergic CR-expressing neurons.