Fig. 2
Gαo is highly targeted to the synaptic terminal regions of cerebellar neurons in wild-type mice. a Gαo-immunoreactivity appears at high levels throughout PC dendrites, especially in Pcp2-positive spines (Arrows in a1-a3, magnified views of a box shown in a). Scale bars, 50 μm in a, 10 μm in a1-a3. b PCs start to differentiate with many protrusions and neurites in a monolayer. Gαo is co-localized with Calb, another PC-specific marker at every PC boundary (yellow, arrows in b2 and b3) except in PC soma devoid of Gαo (asterisks in b2). Scale bars, 20 μm in b-b3. c Strong Gnao mRNA signals appear in the perinuclear regions of PC soma (c1), GCs (c2), and ML interneurons (c3). Scale bars, 20 μm in c, 10 μm in c1-c3. d In vitro culture of PCs shows Gαo and calbindin-D28K (Calb) expression at 21 days in vitro (DIV). The cultured PCs show significant Gαo signal in the soma (asterisk, d2) and in the dendrites (arrows in d1-d2). e In GCs at DIV7, Gαo protein appears in the soma (asterisks, e2) and in the neurites (arrows in e1-e2) of Tubb3-positive GCs. Scale bars, 20 μm in d-e. Bisbenzimide (Hoechst 33342) staining (blue) was used to visualize cell nuclei in a-e