Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 4. Fancd2 opposite-strand (Fancd2os) reduces testosterone production and steroidogenic enzyme expression in TM3 cells. The testoster one levels in Fancd2os-overexpressing (A) or knockdown TM3 cells (B) were detected by enzyme-linked immunosorbent assays (n=3 per group). (C, D, E) Relative quantities of mRNA expression of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) in Fancd2os-overexpressing TM3 cells or Fancd2os knockdown TM3 cells (F, G, H) were determined real-time polymerase chain reaction using β-actin as a housekeeping gene. Each bar represents the mean± standard deviation from three separate experiments. Significant difference compared to TM3, vector/TM3 or NC/TM3. aP<0.05; bP<0.01.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction, Standard Deviation, Plasmid Preparation

Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 6. Higher Fancd2 opposite-strand (Fancd2os) levels in older mouse Leydig cells result in cellular apoptosis and lower serum testosterone production. (A) The serum testosterone levels from mice of different ages were measured using enzyme-linked immunosorbent assays. (B, C) The testis tissues from different aged mice were sliced and then Fancd2os protein expression and apoptosis were analyzed using immuno chemistry and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, respectively. ST represents seminiferous tubule. Black arrows and red arrows indicate Fancd2os-positive cells and TUNEL-positive cells, respectively. Scale bar repre sents 25 μm. The results are given as the mean±standard deviation (n=3). (D) The correlations between Fancd2os expression (B) and the TUNEL-positive staining rate (C) were analyzed using Pearson correlation coefficients. An r≥0.5 was considered to indicate a strong correla tion, and P<0.05 was considered statistically significant. aP<0.05 compared with juvenile mice; bP<0.05 compared with young mice; cP<0.05 compared with middle-aged mice.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, End Labeling, TUNEL Assay, Standard Deviation, Staining

Journal: Cell

Article Title: Zika Virus Causes Testis Damage and Leads to Male Infertility in Mice.

doi: 10.1016/j.cell.2016.11.016

Figure Lengend Snippet: Figure 1. Morphology and Histopathology of the Genital Tracts in Male Mice Infected with ZIKV (A) Morphology of the male genital tract from an uninfected WT mouse. T (testis), SV (seminal vesicle), VP (ventral prostate), VD (vas deferens), EP (epididymis). (B) Morphology of the male genital tract from an uninfected KO mouse. (C) Morphology of the male genital tract from a KO mouse IP infected with ZIKV at 8 DPI (n = 10). Scale bar, 0.5 cm in A, B, and C. The mice from A–C are age matched. (D1) Testis. (E1) Caput epididymis. (F1) Corpus epididymis. (G1) Cauda epididymis. (H1) Ventral prostate. (I1) Seminal vesicle. D1–H1 are representative images from a WT male mouse given PBS by IP (n = 10). (D2) Testis. (E2) Caput epididymis. (F2) Corpus epididymis and (G2) Cauda epididymis. (H2) Ventral prostate and (I2) Seminal vesicle. D2–H2 are representative images from a KO mouse given PBS (n = 10). (D3) Testis. (E3) Caput epididymis. (F3) Corpus epididymis and (G3) Cauda epididymis. (H3) Ventral prostate. (I3) Seminal vesicle. D3–I3 from a WT mouse IP infected with ZIKV (n = 10) at 8 DPI. Abnormalities were not observed. (J) Histological analysis of the testes from ZIKV-infected KO mice. (K) Higher magni- fication of the testis inside the rectangle from D1 (403). Normal seminiferous tubules with different steps of germinal epithelial cells (bigger black arrows) in order and normal intratesticular capillary (white arrow) neighbored with normal testosterone-producing LC (smaller black arrow) in WT testis. (L) Higher magnification of the testis inside the rectangle from D2 (403). Normal seminiferous tubules with different steps of germinal epithelial cells and normal intratesticular capillary (white arrow), neighbored with normal testosterone-producing LC (smaller black arrow) in KO testis. (M) Disrupted seminiferous tubules with cells out of order, different degenerating germinal epithelial cells (bigger black arrows) inside the seminiferous tubules of the ZIKV-infected testes, intratesticular varicoceles/congestion (white arrow), lymphocyte infiltrations (smaller arrows), and necrotic LC (black arrowheads) surrounded by lymphocytes were observed within the interstitium from ZIKV-infected KO testes. (N–P) (N) Caput epididymis. (O) Corpus epididymis and (P) Cauda epididymis. (N–P) Epididymis from ZIKV-infected KO mice. The

Article Snippet: The ELISA kits for IFNb (42400) and for mouse testosterone (DEV 9911) were purchased from R&D Systems (R&D Systems, US) and Demedtec (Kiel-Wellsee, Germany), respectively.

Techniques: Histopathology, Infection