Fig. 6.

Two-color confocal immunofluorescence images of the rat nodulus; cryosections were labeled with cocktails of antibodies to Tbr2, CR and mGluR1α. The cocktail compositions were designed to ascertain whether the sum of the CR⁺ and mGluR1α⁺ UBC subclasses accounts for the entire UBC population. Tbr2 is a marker for all UBC nuclei; CR and mGluR1α are UBC subclass-specific. a The section was immunoreacted with a mixture of all three primary antibodies. The Tbr2 binding sites were revealed with Alexa 594-labeled secondary antibody (red), while binding sites of both CR and mGluR1α were revealed with Alexa 488-labeled secondary antibody (green). All UBCs, whether CR⁺ or mGluR1α⁺ possess Tbr2⁺ nuclei. a’ Distribution of the CR⁺ (c) and mGluR1α⁺ (m) UBCs associated with Tbr2-labeled nuclei. b, b’, b” Enlarged boxed area from panel a illustrates a typical mGluR1α⁺/Tbr2⁺ UBC; intense staining in the brush (arrow) and subtle immunopositivity in soma (arrowhead). c, c’, c’’ Enlarged boxed area from panel a illustrates a typical CR⁺/ Tbr2⁺ UBC; intense staining in the brush (arrow) and cell body (arrowhead). Asterisk indicates Tbr2⁺ UBC nuclei (b’, b”, c’, c”). Scale bars a, a’ 20 μm, b-b’’ and c-c’’10 μm