Semen parameters during ZIKA virus infection in the olive baboon (Papio anubis)

Abstract

OBJECTIVE: Mitochondria are cellular organelles that are required for ATP production, metabolism, and calcium homeostasis, and play a key role in oocyte and embryo development. In this study we aimed to determine whether metabolic imaging using FLIM to detect autofluorescence of NADH and FAD+ (key components of oxidative phosphorylation) can identify metabolic differences between normal oocytes and those with metabolic dysfunction.DESIGN: Experimental study. MATERIALS AND METHODS: Oocytes from old mice (1-year old) (n¼21) were compared to oocytes from young (12-week old) (n¼37) mice as a model of mild oocyte dysfunction. Oocytes obtained from mice with global knockout of Clpp (n¼55) were compared to wild type (WT) oocytes (n¼52) as a model of severe oocyte dysfunction. CLPP (caseinolytic peptidase P) mediates mitochondrial unfolded protein response (mt-UPR) and helps maintain homeostasis in response to metabolic and cellular stress. CLPP is required for oocyte and early embryo development, and ClpP-knockout results in female infertility and accelerated reproductive aging. Cumulus oophorus complexes (COCs) were collected 48h after injection with 5IU pregnant mare serum gonadotropin (PMSG, Sigma), and germinal vesicle (GV) oocytes were isolated. Fluorescence lifetime imaging microscopy (FLIM) was used to measure the naturally occurring autofluorescence of NADH and FAD+ in individual oocytes. A total of 8 metabolic parameters were obtained from each measurement (4 for each fluorophore) that are sensitive to differences in metabolic state: short (T1) and long (T2) fluorescence lifetime, fluorescence brightness (IRR), and fraction of the molecule bound to enzyme (FB). Matlab was used for statistical analysis.RESULTS: In older oocytes compared to young ones, FAD+ short fluorescence time (T1) was longer (p<0.001) and fluorescence brightness (IRR) was lower (p<0.01), while NADH short and long fluorescence times (T1 and T2) were shorter (p<0.0001 for both), and brightness (IRR) and fraction bound (FB) were lower (p<0.0001 and p<0.05, respectively). In ClpP-knockout oocytes compared to WT, FAD+ short and long fluorescence times (T1 and T2) were longer (p<0.0001) and brightness (IRR) was higher (p<0.01), while NADH long fluorescence time (T2) was longer (p<0.0001), and fraction bound (FB) was smaller (p<0.0001).CONCLUSIONS: In this study we used metabolic imaging to characterize the metabolic state of oocytes with mild (old) and severe (ClpP-knockout) e52

From Ancient to Emerging Infections: The Odyssey of Viruses in the Male Genital Tract

“…In ZIKVinfected NHPs (287,364,390,524), the virus was occasionnally detected in the testis after day 7-8 post-infection (364,390,524), without tissue alteration, and scarcely infected cells were observed in the prostate and seminal vesicles (524), along with low levels of viral RNA similar to other non MGT tissues (287). In 3/6 infected baboons, a marked transient decrease in total and motile sperm count was observed, but these differences did not reach significance (154). Overall, the drastic effect of ZIKV on the MGT in mice significantly differs from that in ZIKVinfected men and NHP, in whom orchitis has never been reported and modifications of sperm parameters appear milder and transient.…”