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Additional file 6 of PA1 participates in the maintenance of blood–testis barrier integrity via cooperation with JUN in the Sertoli cells of mice

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posted on 2022-04-05, 08:33 authored by Bo Liu, Chao Liu, Binfang Ma, Ruidan Zhang, Zhiwei Zhao, Sai Xiao, Wanjun Cao, Yanjie Ma, Guozhang Zhu, Wei Li, Zhen Li
Additional file 6: Fig. S4. Amh-pa1−/− mice show massive germ cell loss. a) Most of the residual cells in the empty tubules of Amh-pa1−/− testis were identified as Sertoli cells. Immunofluorescence analysis using anti-WT1 (green) was conducted on the sections of testis from Pa1F/F mice (upper panels) and Amh-pa1−/− mice (lower panels). Nuclei were stained with DAPI (blue). Dashed curve indicated basal membrane of seminiferous tubules. b) PCNA immunofluorescence analysis of seminiferous tubules showed that 8-week-old Amh-pa1−/− mice had decreased numbers of spermatocytes and spermatogonia compared with 8-week-old Pa1F/F mice. Seminiferous tubule sections of Pa1F/F and Amh-pa1−/− testes were stained for PCNA (red), and the nuclei was stained with DAPI (blue). Dashed boxes showed the localization of the enlarged images. Dashed curve indicated basal membrane of seminiferous tubules. Data are presented as mean ± SEM. ***p < 0.001. c) DDX4 immunofluorescence analysis of germ cells showed that 8-week-old Amh-pa1-/- mice had decreased numbers of germ cells compared with 8-week-old Pa1F/F mice. Seminiferous tubule sections of Pa1F/F and Amh-pa1−/− testes were stained for DDX4 (red), and the nuclei was stained with DAPI (blue). Dashed boxes showed the localization of the enlarged images. Dashed curve indicated basal membrane of seminiferous tubules. Data are presented as mean ± SEM. **p < 0.01. d) PLZF immunofluorescence analysis of germ cells showed that 8-week-old Amh-pa1−/− mice had decreased numbers of undifferentiated spermatogonium compared with 8-week-old Pa1F/F mice. Seminiferous tubule sections of Pa1F/F and Amh-pa1−/− testes were stained for PLZF (red), and the nuclei was stained with DAPI (blue). Dashed boxes showed the localization of the enlarged images. Dashed curve indicated basal membrane of seminiferous tubules. Arrows indicated PLZF positive cells. e) PCNA positive cell number per tubule was significantly decreased Amh-pa1−/− (27.34 ± 1.319) mice testes compared with Pa1F/F mice (62.72 ± 3.167). (n = 3 independent experiments). Data are presented as mean ± SEM. ***p < 0.001. f) DDX4 positive cell number per tubule was significantly decreased in Amh-pa1−/− (50.39 ± 2.342) mice compared with Pa1F/F mice (122.9 ± 12.20). (n = 3 independent experiments). Data are presented as mean ± SEM. **p < 0.01. g) PLZF positive cell number per tubule was significantly decreased Amh-pa1−/− (0.5789 ± 0.1919) mice testes compared with Pa1F/F mice (2.763 ± 0.2321). (n = 3 independent experiments). Data are presented as mean ± SEM. ***p < 0.001. h) Homologous chromosome synapsis, pairing, or segregation were normal undergoing in Amh-pa1−/− spermatocytes. Spermatocytes of Pa1F/F and Amh-pa1−/− mice were stained with SYCP3 (green) and γH2AX (red) antibodies.

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National Natural Science Foundation of China

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