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. 2022 Feb 17;13:914. doi: 10.1038/s41467-022-28611-2

Fig. 7. Role of BDNF-TrkB in the human ovary.

Fig. 7

a Effect of Ab4B19 on the proliferation of human KGN cells. KGN cells were cultured for 72 h and then treated with Ab4B19 for 24 h. Cell proliferation was measured by the viability analysis on 96-well plates after treatment with BDNF (as a positive control, 0.2 nM) and Ab4B19 (0.2, 0.6, or 1.8 nM) for 48 h (N = 5 per group). b Activation of CREB by Ab4B19 in KGN cells. Cultured KGN cells were treated with BDNF (0.2 nM) and Ab4B19 (0.2, 0.6, or 1.8 nM) for 24 h, and harvested for pCREB assay. Phosphorylated CREB (Ser133) and total CREB were examined by Western blot. Quantification of pCREB/CREB was plotted on the right. The levels of vehicle treatment were normalized to 1. N = 3 repetitions per condition. c–e Transcriptional analysis of the expression of BDNF and TrkB in human GCs and oocytes. The raw data were derived from Zhang et al. (Zhang, Yaoyao, et al. Molecular Cell 72.6 (2018): 1021-1034.). Bdnf and TrkB genes with fragments per kilobase per million (FPKM) >1 in at least one cell were analyzed, and expression levels of each gene were plus one then log2 transformed in the following analysis. c Bar plots showing the relative expression levels (log2 [FPKM + 1]) of BDNF mRNA from the primordial to preovulatory ovarian follicle stages. d Expressions of TrkB-T1 and TrkB-FL mRNAs in human GCs from five different developmental stages of ovarian follicles. e Expressions of TrkB-T1 and TrkB-FL mRNAs in human oocytes from five different developmental stages of ovarian follicles. f Typical image of BDNF expression in the human ovary from young and aged people. BDNF-immunoreactivity was detected by a rabbit anti-BDNF antibody followed by anti-rabbit IgG (FITC, green). Co-labeling with a mouse anti-VASA antibody followed by anti-mouse IgG (TRITC, red) was performed to identify follicles. The outlines of follicles were marked using dotted white circles. Scale bar, 100 μm. g Relative fluorescence intensity of BDNF expression in young (29–35 years old) and aged (61–64 years old) groups. (N = 4 people in the young group and 3 people in the old group). Data were all presented as mean ± SEM. Statistical analyses were carried out using one-way ANOVA followed by the Dunnett’s multiple comparisons test (a, b), or two-tailed student’s t-test (g), respectively (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). Source data are provided as a Source Data file.