Figure 5.

Reduced glutamatergic innervation of kisspeptin neurons in HFD. A, AMPA receptors were quantified in kisspeptin neurons in CRL and HFD. Top, representative images of GluR2 (red) and VGLUT2 (green) colocalized to kisspeptin neurons (white, 630× magnification). Bottom, 3D reconstruction using Imaris to assure counting cell surface receptors in direct contact with kisspeptin neurons. B, Quantification of GluR2 (top) and VGLUT2 (bottom) in kisspeptin neuron soma (left) and proximal 15 μm of the process (right). Each dot represents an average for one mouse (15–40 neurons counted per animal), and bars represent the group average for CTR (gray) and HFD (red). C, Fewer NMDA receptors on kisspeptin neurons in HFD mice. Top, confocal and bottom, 3D reconstructed representative images of postsynaptic NR2A (red) and presynaptic VGLUT2 (green) on kisspeptin neurons (white). D, Quantification of NR2A numbers in CTR and HFD at the soma and 1–15 µm proximal process. E, Quantification of GluR2 and VGLUT2 colocalization at the kisspeptin neuron soma and proximal 1–15 µm process in CTR or HFD. F, Quantification of NR2A and VGLUT2 colocalization at the kisspeptin neuron soma and proximal 1–15 µm process in CTR or HFD. Each dot represents an average for one mouse (15–40 neurons counted per animal), and bars represent the group average for CTR (gray) and HFD (red). Statistical significance between CTR and HFD is indicated with a * (p < 0.05).