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. 2020 Jul 9;9:e53945. doi: 10.7554/eLife.53945

Figure 3. Chemogenetic inhibition of GnRH neuron distal dendron activity suppresses both the surge and pulse profiles of LH secretion.

(A) Schematic showing experimental protocol with GnRH-Cre mice injected with Cre-dependent Flex-hM4D(Gi)-mCherry AAVs bilaterally into the region of the median eminence and CNO given into the same region. MS, medial septum. CNO, Clozapine N-oxide. (B) Fluorescence images of GnRH neuron projections in the vicinity of the median eminence expressing GnRH (green) and mCherry (red) in GnRH::Flex-hM4D(Gi)-mCherry mice. Scale bar, 20 μm. (C,D) LH surge profiles in all GnRH-Cre OVX+E+P female mice following MBH injection of saline (C, n = 12) or CNO (D, n = 12). (E) Mean (± SEM) LH levels. ***p<0.001, two-way repeated measures ANOVA with Holm-Sidak test. F-J. Representative examples of pulsatile LH secretion in OVX GnRH-Cre mice given saline (F,G) or CNO (I,J). LH pulses are indicated by asterisks. Mean (± SEM) amplitude (H) and frequency (K) of LH pulses in saline (n = 8) and CNO (n = 7). ***p<0.001,*p<0.05 Mann-Whitney U-tests.

Figure 3.

Figure 3—figure supplement 1. Photomicrographs showing location of cannula and mean (± SEM) LH levels in GnRH-Cre OVX mice with saline and CNO.

Figure 3—figure supplement 1.

(A) Photomicrograph showing location of cannula implanted into the MBH of GnRH::Flex-hM4D(Gi)-mCherry mice. Scale bar, 100 μm. Mean (± SEM) LH levels in AAV-injected GnRH-Cre OVX mice with IP injection of saline (B, n = 6) or CNO (C, n = 6). Mean (± SEM) LH levels in AAV-injected GnRH-Cre OVX mice with MBH injection of saline (D, n = 8) or CNO (E, n = 7). Mean (± SEM) LH levels in AAV-injected GnRH-Cre OVX mice with rPOA injection of saline (F, n = 8) and CNO (G, n = 9).