Figure 2.

GnRH Induces Pyk2 Phosphorylation in Vivo

Mature ovariectomized mice received anti-GnRH antiserum (250 μl) ip to immunoneutralize endogenous GnRH. Three days later, immunoneutralized mice received either saline or an analog of GnRH (d-Ala⁶-GnRH; 100 ng ip) and were killed 45 or 90 min later. Pituitaries were dissected out, fixed, sectioned, and examined for Pyk2 (pY⁸⁸¹) phosphorylation or LHβ protein by double-labeled fluorescent immunocytochemistry. The first column depicts labeling for LHβ (green), the second column of images phospho-Pyk2 (pY⁸⁸¹; red), and the final column of images is a merge of the two images. The first row depicts results from cells collected from mice receiving saline for 60 min (control). The second and third rows of images depict cells collected 45 and 90 min (respectively) after d-Ala⁶-GnRH administration. The fourth row of images is immunocytochemical analysis using normal rabbit serum substituted for the phospho-Pyk2 antibody on tissues collected at 90 min after d-Ala⁶-GnRH treatment. The figure reflects a representative example from three or four mice per treatment. n, Nucleus. Bar, 5 μm.