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. 2024 Apr 30;15:3610. doi: 10.1038/s41467-024-47300-w

Fig. 3. Exposure to pubertal stress did not affect RP3V kisspeptin neurons and their activation by male odors whereas it reduced the number of the nNOS neurons in the VMHvl and their activation by male odors.

Fig. 3

a Schematic representation of the protocol used to induce Fos expression in the neuronal circuit involved in sexual behavior. Figure created with BioRender.com. b Representative images of double labeling for kisspeptin and Fos in the RP3V following exposure to clean versus male bedding. Black arrows show examples of kisspeptin neurons, red arrows show examples of Fos expressing neurons, while pink arrows point towards examples of neurons coexpressing kisspeptin and Fos. c Number of kisspeptin neurons in the control (n = 4) and pubertal stress group (n = 5) groups following exposure to clean bedding, and in the control (n = 7) and pubertal stress (n = 7) after exposure to male bedding. Two-way ANOVA; Bedding effect: F(1, 16) = 4.968, p = 0.0381; group effect: F(1,19) = 0.177, p = 0.679; bedding × group effect: F(1,19) = 0.39, p = 0.539. d Percentage of neurons coexpressing kisspeptin and Fos in the control (n = 4) and pubertal stress (n = 5) groups following exposure to clean bedding, and in the control (n = 7) and pubertal stress (n = 7) after exposure to male bedding. Two-way ANOVA; Bedding effect: F(1,19) = 23.15, p = 0.0001; group effect: F(1,19) = 2.7778, p = 0.1119; interaction: F(1,19) = 0.13, p = 0.722. e Representative confocal images showing nNOS/Fos immunofluorescence staining in the VMHvl. White dashed boxes indicate the regions of high magnification (f) Number of nNOS-immunoreactive neurons in the VMHvl of the control and pubertal stress groups following exposure to clean or male bedding. Two-way ANOVA: bedding effect: F(1,18) = 0.068, p = 0.796; group effect: F(1,18) = 16.28, p = 0.0008); bedding × group: F(1, 18) = 0.1088, p = 0.745. ***p < 0.001. g Percentage of nNOS neurons expressing Fos following exposure to male bedding. Two-way ANOVA followed by Bonferroni multiple comparison test; Bedding effect: F(1,18) = 51.23, p < 0.0001; Group effect: F(1,18) = 4.684, p = 0.044; interaction: F(1,18) = 5.419, p = 0.0318. *p < 0.05; ***p < 0.001. n = 4 for control group exposed to clean bedding, n = 5 for pubertal stress group exposed to clean bedding, n = 7 for control group exposed to male bedding, n = 6 for pubertal stress group exposed to male bedding. All data are presented as mean ± SEM. Corrections were made whenever multiple comparisons test is used. Source data are provided as a Source Data file.