Figure 4

Immunofluorescent localization of Ce-GnRHR to the germline. (i). Permeabilized adult N2 worm probed with an anti-human GnRHR1 monoclonal antibody. Anterior view illustrating apparent nuclear immunostaining of maturing oocytes, and sperm. (ii). Brightfield image of same worm shown in (i). (iii). Immunofluorescent localization of GnRHR in a permeabilized adult worm using a polyclonal antibody raised against Ce-GnRHR. High magnification image of anterior gonadal arm (highlighted in white) illustrates nuclear staining of maturing oocytes, and intestinal (yellow arrows) cells. Immunofluorescent staining clearly illustrates an increase in the size of the oocyte nuclei as they mature along the gonadal arm. (iv). Permeabilized adult worm probed with rhodamine-coupled secondary antibody (control for anti-human GnRHR1 antibody staining). (v). Permeabilized adult worm probed with FITC-coupled secondary antibody (control for anti-Ce-GnRHR antibody staining). No specific immunoreactivity was detected with either secondary antibody alone. Scale (i, ii): 100 μm., Scale (iii-v): 50μm.