Figure 4

In situ localization of CART mRNA within granulosa cells of dominant ovulatory versus nonovulatory (subordinate) bovine follicles collected before (0 h) and after (12 h) GnRH injection to induce the preovulatory gonadotropin surge (n = 3 each). A) Bright field micrograph of section through dominant ovulatory follicle (denoted as DF) and adjacent non-ovulatory subordinate follicle (denoted as SF) collected before the preovulatory gonadotropin surge (0 h timepoint) and hybridized with ³⁵S-antisense CART cRNA. B) Bright field micrograph of section through DF and adjacent SF collected after the preovulatory gonadotropin surge (12 h timepoint) and hybridized with ³⁵S-antisense CART cRNA. C) Dark field micrograph of section adjacent to one depicted in Panel A. D) Dark field micrograph of section adjacent to one depicted in Panel B. Note localization of CART mRNA to GC of the SF (right) but not the DF (left) depicted in Panels A-D. E) Magnified bright field micrograph of SF depicted in Panels A and C. F) Magnified bright field micrograph of SF depicted in Panels B and D. G) Magnified dark field micrograph of Panel E. H) Magnified dark field micrograph of Panel F. Note localization of CART mRNA specifically to GC of SF in panels G and H. I and J) Dark field micrographs of sections adjacent to ones depicted in Panels E and F hybridized with ³⁵S-sense CART cRNA (negative control). A-D: magnification = 100X, scale bar = 200 μm; E-J: magnification = 400X, scale bar = 50 μm. Granulosa cell layer denoted as GC. Theca cell layer denoted as TC. Reproduced from Kobayashi et al., 2004 [59].