Fig. 4.

Wnt1 is a novel actor in astrocyte-neuron crosstalk. A: scheme of ventral midbrain astrocytes (VM-AA) isolation, purification and direct (co-culture) or indirect (AS-conditioned medium, ACM) culture paradigms with purified primary mesencephalic dopaminergic neurons (mDAn), in the absence or the presence of the PD neurotoxin, MPP⁺. Survival, dopamine (DA) uptake and caspase 3 assays are used to monitor mDAn death and functionality. B:In situ hybridization histochemistry coupled to confocal laser scanning microscopy and dual immunofluorescent staining with the astrocyte cytoskeleton marker, glial fibrillary acidic protein (GFAP, in red) and Wnt1 mRNA (in green) showing the expression of Wnt1 in primary GFAP⁺ cells (orange-yellow). C: Astrocyte-neuron crosstalk with primary mDAns. Confocal image of TH⁺ neurons (in green) in co-culture with VM-AS (in red), showing TH⁺ neurons with long and branched TH⁺ neuronal processes, interacting with GFAP⁺ star-shaped astrocytes. Fig. 4. Scale bars, panel B: 25 μm, Box: 10 μm, panel C: 50 μm.