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. Author manuscript; available in PMC: 2014 Mar 14.
Published in final edited form as: Nat Neurosci. 2008 May 18;11(6):634–636. doi: 10.1038/nn.2126

Figure 1.

Figure 1

Origin, targets and androgen dependence of GRP neurons in the lumbar spinal cord. (a–d) More GRP-immunoreactive neurons were found in upper lumbar spinal cord of WT males (a) than females (b). Tfm males were feminine in this regard (c), with significantly fewer GRP+ neurons in this region than WT females (d; means ± s.e.m.). (e–h) In WT males, nearly every GRP-immunoreactive neuron also contained androgen receptor (e,f), but these neurons did not contain ERα (g,h). (i–l) In lower lumbar spinal cord autonomic nuclei, males (i) had more GRP-immunoreactive fibers in the SPN (red inset) and the DGC than did females (j; quantitation in l). Androgen receptor–lacking Tfm males were feminine in this regard (k). GRP fibers in the non-autonomic dorsal horn region of the spinal cord (DH) were equivalent across groups (l). (m–u) GRP-containing fibers closely apposed cell bodies and proximal dendrites of nNOS+ neurons in the SPN. Left column, WT male; center, WT female; right, Tfm male. (v) Possible synaptic sites (arrowheads). (w) Electron micrographs confirmed GRP-immunoreactive presynaptic boutons innervating nNOS-containing dendrites in the SPN. Arrows, GRP-immunoreactive secretory granules; arrowheads, synapse. GRP-t, GRP terminal; m, mitochondrion; Den, nNOS-immunoreactive dendrite. *P < 0.001 compared with WT males; †P < 0.001 compared with WT females (one-way (d) or two-way (l) analysis of variance; significant effects followed by post hoc Bonferroni tests). Scale bars: c (also applies to a,b) and g (applies to e), 100 μm; h (applies to f), 10 μm; k (applies to i,j), 200 μm; u (applies to m–t), 50 μm; v, 10 μm; w, 200 nm. All animal procedures were approved by institutional review boards of Kyoto Prefectural University of Medicine and/or Michigan State University. For methodological details, see Supplementary Methods online.