Figure 8.

GnRH induces ERK1/2-dependent bleb formation in primary cultures of rat pituitary cells and isolated mouse gonadotropes. (A) DIC images from a confocal microscope of dissociated rat pituitary cells. Culture rat pituitary cells were prepared, and gonadotropes were identified by their Ca²⁺ response as described in Section “Materials and Methods.” The cells were treated with GnRH (10 nM) for 30 min. (B) Quantitation of the percentage of blebbing cells from dissociated rat pituitary cells. The cells were pretreated with U0126 (10 µM) for 30 min, followed by GnRH (10 nM) for additional 30 min. Data are mean ± SEM from three experiments. (C) DIC and fluorescent images from a confocal microscope of FACS-purified primary gonadotrope cells from adult GRIC/Ai9 mice treated with GnRH (30 min, 10 nM). The scale bar is 5 µm. (D) Images from a confocal microscopy time-lapse movie of FACS-purified primary gonadotrope cells. The cells were pretreated with or without U0126 (25 µM) for 30 min, followed by GnRH (10 nM) for additional 30 min. The scale bar is 5 µm. (E) Quantitation of the percentage of blebbing cells from panel (D) is presented (at least 30 cells for each experiment) and data are mean ± SEM. **p-value ≤0.01.