ABSTRACT
Spermatogenesis is a complicated process involving mitotically proliferating spermatogonial cells, meiotically dividing spermatocytes, and spermatid going through maturation into spermatozoa. The post-translational modifications of proteins play important roles in this biological process. S-palmitoylation is one type of protein modifications catalyzed by zinc finger Asp-His-His-Cys (ZDHHC)-family palmitoyl S-acyltransferases. There are 23 mammalian ZDHHCs that have been identified in mouse. Among them, Zdhhc19 is highly expressed in adult testis. However, the in vivo function of Zdhhc19 in mouse spermatogenesis and fertility remains unknown. In this study, we knocked out the Zdhhc19 gene by generating a 2609 bp deletion from exon 3 to exon 6 in mice. No differences were found in testis morphology and testis/body weight ratios upon Zdhhc19 deletion. Spermatogenesis was not disrupted in Zdhhc19 knockout mice, in which properly developed TRA98+ germ cells, SYCP3+ spermatocytes, and TNP1+ spermatids/spermatozoa were detected in seminiferous tubules. Nevertheless, Zdhhc19 knockout mice were male infertile. Zdhhc19 deficient spermatozoa exhibited multiple defects including abnormal morphology of sperm tails and heads, decreased motility, and disturbed acrosome reaction. All of these led to the inability of Zdhhc19 mutant sperm to fertilize oocytes in IVF assays. Taken together, our results support the fact that Zdhhc19 is a testis enriched gene dispensable for spermatogenesis, but is essential for sperm functions in mice.
Subject(s)
Acyltransferases/physiology , Fertilization , Sperm Motility , Spermatocytes/cytology , Spermatogenesis , Spermatozoa/physiology , Acrosome Reaction , Animals , Female , Male , Mice , Mice, Knockout , Spermatocytes/physiologyABSTRACT
Increased production of reactive oxygen species (ROS) and inflammation are major contributors to the development and progression of diabetes-associated erectile dysfunction (DMED). As an endogenous antioxidant and anti-inflammatory factor, the potential implication of pigment epithelium-derived factor (PEDF) in DMED has not been revealed. To assess the potential antioxidant and anti-inflammatory functions of PEDF in DMED, we first demonstrated that PEDF was significantly decreased at the levels of the mRNA and protein in the penis of diabetic rats compared with normal controls. To test the hypothesis that decreased the penile levels of PEDF are associated with oxidative stress and inflammation in DMED, an adenovirus expressing PEDF (Ad-PEDF) or the same titer of control virus (Ad-GFP) was intracavernously administered at 2 weeks after diabetic onset. After 6 weeks of treatment, we found that administration of Ad-PEDF could significantly increase erectile response to cavernosal nerve stimulation in the diabetic rats by restoring the endothelial NO synthase (eNOS), P-eNOS, and neuronal NO synthase (nNOS) protein levels to the standard levels represented in normal rats and by suppressing the levels of tumor necrosis factor-α (TNF-α) and oxidative stress. In conclusion, the present data indicated that the antioxidant and anti-inflammatory potential of PEDF plays important role in restoring erectile function by the inhibition of oxidative stress and TNF-α production.
Subject(s)
Diabetes Mellitus, Experimental/genetics , Eye Proteins/genetics , Nerve Growth Factors/genetics , Penile Erection/genetics , Penis/metabolism , Serpins/genetics , Animals , Diabetes Mellitus, Experimental/metabolism , Down-Regulation , Eye Proteins/metabolism , Gene Expression Regulation , Male , Nerve Growth Factors/metabolism , Nitric Oxide Synthase Type I/metabolism , Nitric Oxide Synthase Type III/metabolism , Oxidative Stress , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Serpins/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
There are some defects in constructed wetland, including the uneven distribution of flow, easily blocked, lack of oxygen supply systems and the unsatisfactory phosphorus adsorption capacity of the substrates, etc. The research mainly studied the function and contribution of the pool of hydrolysis acidification, the natural reoxygenation/sinking device and strengthen slot for reducing nitrogen and phosphorus. The results showed the removal efficiency of COD and SS in hydrolysis-acidification pool accounted for 38.05% and 34.82% of the total removal efficiency of system. The SS removal efficiency of the natural reoxygenation/sinking device accounted for 22.01% of the total removal efficiency of system, and the concentration of DO kept above 2.5 mg.L-1. The two strengthen slots can ensure the TP, TN, COD up to the standard of the level 1 of GB 18918-2002.
