Figure 1.
Confocal microscopy revealing GFP-expressing GnRH neurons (left) and immunostaining with LR1 (right).
Note that the two techniques reveal a similar number of GnRH perikarya, but the extent of the neurites is clearer using the immunohistochemical approach. Scale bars = 50 µm.
Figure 2.
Confocal microscopy photomicrographs of 0.5 µm optical sections showing the relationship between a vasoactive intestinal polypeptide fiber (A: VIP, red) and a GnRH-GFP perikaryon (B: GnRH, green). Panel C shows the superimposed images. Scale bars = 20 µm.
Table 1.
Summary of peptidergic appositions on GnRH-GFP perikarya.
Figure 3.
A: NPY-ir perikarya localized in the arcuate nucleus. Examples of close appositions between NPY-ir fibers (red) and GnRH-GFP perikarya and processes (green) in a maximum intensity projection of the image stack (B) and in single 0.5 µm optical slices (C, D). NPY-ir fibers and GnRH-ir terminals were observed in proximity in the median eminence (E), where the presence of close appositions was confirmed by analysis of single 0.5 µm optical slices (F). Scale bars = 50 µm (A), 10 µm (B–D), 50 µm (E), 5 µm (F).
Figure 4.
Examples of close appositions between β-endorphin-ir fibers (red) and GnRH-GFP perikarya and processes (green) in a maximum intensity projection of the image stack (A) and in single 0.5 µm optical slices (B–D).
A few close appositions were also observed between β-endorphin-ir fibers and GnRH neuroterminals in the median eminence (E), confirmed by analysis of single 0.5 µm optical slices (F). Scale bars = 15 µm (A–D), 10 µm (E), 5 µm (F).
Figure 5.
A: MCH-ir perikarya localized in the lateral hypothalamic area (A). Examples of close appositions between MCH-ir fibers (red) and GnRH-GFP perikarya and processes (green) in a maximum intensity projection of the image stack (B) and in single 0.5 µm optical slices (C, D). MCH -ir fibers and GnRH-ir terminals were observed in proximity in the median eminence (E), where the presence of close appositions was confirmed by analysis of single 0.5 µm optical slices (F). Scale bars = 25 µm (A), 5 µm (B–D), 10 µm (E), 5 µm (F).