Zika virus causes testicular atrophy

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Science Advances  22 Feb 2017:
Vol. 3, no. 2, e1602899
DOI: 10.1126/sciadv.1602899


  • Fig. 1 Characterization of ZIKV in mice.

    Ifnar1−/− mice (n = 4) were infected with 105 plaque-forming units (PFU) of ZIKVMEX via subcutaneous route. (A) Weights were monitored for 15 days after virus infection. The values represent average scores of body weight compared with initial body weight ± SD from four mice. (B) Blood samples were collected at 1, 3, 5, 7, 9, and 21 dpi, and the virus loads were examined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Three wild-type (WT) and Ifnar1−/− mice were infected subcutaneously with 105 PFU of ZIKVMEX. Multiple organs were collected from mice (n = 3) after challenge on day 5 and subjected to virus loads by qRT-PCR (C) and by plaque assays in Vero cells (D). The results are expressed as the mean titers. The detection limit of plaque assays is 1.8 log10 PFU/g.

  • Fig. 2 ZIKV infection in testes and epididymis.

    Ifnar1−/− mice were infected with 105 PFU of ZIKVMEX via subcutaneous route. (A) IHC was performed using an anti-ZIKV NS1 antibody. Left panels (low magnification) show insets shown on the right (higher magnification). Red arrowheads indicate infected epithelial cells. (B and C) At 5 dpi, testes were collected, and total RNA was extracted for qRT-PCR of interferon-β (IFN-β), tumor necrosis factor–α (TNF-α), interleukin-6 (IL-6), IFN-γ, and interferon-stimulated gene 15 (ISG-15) (B). The testicular mRNA expression levels of testosterone-synthetic genes [steroidogenic acute regulatory factor (StAR), cytochrome P-450 side-chain cleavage enzyme (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD)] were examined by qRT-PCR (C). Data are normalized to mouse β-actin (*P < 0.05). (D) Isolated Leydig cells from WT and Ifnar1−/− mice were infected with ZIKV at a multiplicity of infection of 10. At the indicated times after infection, virus titers in the supernatant were determined with Vero cells. The reported values are means ± SD.

  • Fig. 3 Testicular abnormalities after ZIKV infection.

    The testes of Ifnar1−/− mice infected with 105 PFU of ZIKVMEX via subcutaneous route and of mock-infected mice were harvested at 5 and 21 dpi. (A) The pictures compare the testes of infected mice. Scale bar, 1 cm. (B and C) Weight and length of whole testes from infected animals were measured (n = 6 testes comprising three mice; **P < 0.01). Same color indicates the same mice. (D) The levels of testosterone in blood of infected mice were investigated by enzyme-linked immunosorbent assay (*P < 0.05). The detection limit is 0.3 ng/ml. (E) The relative viral RNA levels in testes and epididymis at 5 and 21 dpi were examined by qRT-PCR. Data are normalized to mouse β-actin.

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