Maturation of sperm volume regulation in the rat epididymis

Sperm maturation in the epididymis may involve differences between mature and immature spermatozoa in their volume regulatory osmolyte response. Spermatozoa obtained from the rat caput and cauda epididymidis were examined for their ability to regulate volume after transfer from in situ epididymal osmolality （measured to be 343 ± 13 and 365 ± 19 mmol kg^-1, respectively） to that of the female tract in single- and multiple-step protocols. Cells withstood the single-step treatment better than the multistep protocol. Sperm volume estimates by flow cytometric measure- ments of forward scatter of cells with intact head membranes was more sensitive than those by assessing cell coiling microscopically. At osmolalites below 210 mmol kg l both caput and cauda cells ruptured, limiting the use of flow cytometry. Above this critical value, the use of quinine showed that both caput and cauda cells could regulate volume, but cauda cells were the more effective. Of several organic osmolytes studied, myo-inositol, glutamate and KCl caused only temporary and slight swelling of spermatozoa cells in hypotonic medium. Spermatozoa of both maturities seemed to use potassium as the preferred osmolyte for regulating volume.

Human chorionic gonadotropin in vitro: Effects on rat sperm motility and fertilization outcome

Objective:To investigate the effect of human chorionic gonadotropin (hCG) on the motility of rat sperm and the fertilization rate following in vitro fertilization. Methods: hCG concentrations of 25, 50, 75 and 100 ng/mL with incubation time of 1 h and 2 h were used in medium to study sperm motility. Then, 25 and 100 ng/mL of hCG with incubation time of 2 h were applied for in vitrofertilization. After 6 h, the number of two pronuclei was counted. Obtained data was subjected to one way ANOVA and Bonferroni-Dun hoc-post test.Results:Total motility and progressive motility of sperms were increased significantly (P<0.05) in hCG experimental group with concentration of 25 ng/L and incubation time of 2 h compared to control group, however, total motility and progressive motility of sperms were decreased significantly (P<0.05) with increasing dose of hCG. Fertilization rate was decreased significantly (P<0.05) in hCG experimental group with dose of 100 ng/mL compared to 25 ng/mL and control group. Conclusions:The effect of hCG on the rat sperm motility and the rate of their fertility is dose-dependent so that hCG with dose of 25 ng/mL leads to an increase in rat sperm motility and to some extent fertilization rate.

Effect of Cadmium on Rat Leydig Cell Testosterone Production and DNA Integrity in vitro

Cadmium （Cd） is an elemental heavy metal with widely recognized toxicity. Its long-term use in industrial processes and daily activities has caused alarming levels of Cd contamination in the natural environment. According to the estimates by the Agency of Toxic Substances and Disease Registry in the US, 25 000 to 30 000 metric tons of Cd is annually released to the environment . Results of previous studies have demonstrated that several organs are targets of Cd, but the most important of these targeted organs may be the testes.

The reversibility of sperm quality after discontinuing nandrolone decanoate in adult male rats

Aim： To investigate the reversibility of the effect of nandrolone decanoate （ND） on sperm parameters after discontinuing the drug. Methods： Three groups of rats received peanut oil （control）, low and high doses of ND for 14 weeks, respectively. Each group was divided into subgroups A and B, in which rats were killed 14 and 28 weeks after initiating the injection, respectively. Results： Sperm count and motile sperm fraction were decreased in the subgroups A and B that received low and high doses of ND in comparison with the controls A and B, respectively. The sperm count and motile sperm fraction increased in the subgroups B that received low and high doses of ND in comparison with their corresponding subgroups A. The number of normal morphology sperm was decreased significantly in subgroups A receiving low and high doses of ND in comparison with the control subgroup A. However, this parameter was not decreased in subgroups B receiving low and high doses in comparison with the control subgroup B. The normal sperm morphology did not show any significant differences in the subgroups B in comparison with their corresponding subgroups A. Conclusion： The 14-week injection of low and high doses of ND decreases sperm quality and quantity in rats. These parameters were improved after discontinuing ND, but not recovered completely even when they are left untreated for 14 weeks.

Plasma antioxidant capacity, sexual and thyroid hormones levels, sperm quantity and quality parameters in stressed male rats received nano-particle of selenium

Objective:To evaluate the effects of nano-particle of selenium (nSe) on plasma antioxidant capacity, sexual and thyroid hormones and spermatogenesis in male rats exposed to oxidative stress.Methods:Forty rats were randomly divided into four treatments with ten replicates. Treatment groups were: C, the control group received normal saline as gavage and injection (i.p.);OS, received tert-butyl hydroperoxide (0.2 mmol/kg body weight) for inducing oxidative stress;nSe, received nSe (0.3 mg/kg body weight) as gavage, and OS+nSe, received tert-butyl hydroperoxide and nSe. All groups were treated for 28 d and administrations were done each 48 h.Results:Oxidative stress decreased and gavage of nSe to stressed rats increased the antioxidant capacity and activities (P<0.05). Oxidative stress decreased and nSe increased the levels of thyroid, gonadotropic and testosterone hormones as compared with the control group (P<0.05). Gavage of nSe to stressed rat resulted in increase (P<0.05) of gonadotropin and testosterone hormones compared to oxidative stress group. There were no significant differences (P>0.05) between rats exposed to oxidative stress and those in the control group for sperm quantity and quality. Gavage of nSe to stressed rats had no effect (P>0.05) on the sperm parameters, except increased viability and progressive percentages.Conclusions:Nano-particle of Selenium administration in stressed rats could ameliorate the negative effects of oxidative stress on the antioxidant capacity and activities, but not on the quantity and quality parameters of sperm.

Protective effects of honey compound syrup on busulfan-induced azoospermia in male rats

Objective:To evaluate the protective effects of honey compound syrup on sperm count and testis tissue in rats.Methods:Thirty rats were randomly assigned to five groups.The control group received 1 mL normal saline with dimethyl sulfoxide intraperitoneally;the busulfan group received busulfan 10 mg/kg body weight at the first and twenty-first days of the experiment via intraperitoneal injection;the last three groups received busulfan 10 mg/kg body weight to induce azoospermia,and then received 1.0,1.5,or 2.0 mg/kg honey compound syrup,respectively,after induction of azoospermia.After administration,the testis and epididymis of all rats were removed.Then,reproductive organ weight and sperm parameters(sperm concentration,epididymal sperm reserve and daily sperm production)were measured.After hematoxylin-eosin staining,seminiferous tubule cells and diameters were assessed.Results:Busulfan damaged the testis tissue and impaired spermatogenesis.Administration of honey compound syrup in three doses improved testis tissue and spermatogenesis.The protective effects of honey compound syrup may relate to the antioxidant properties of honey and other compounds in this syrup.Conclusions:Administration of honey compound syrup could be an ameliorative agent for the side effects of chemotherapy drugs such as busulfan on the male reproductive system.

Nutritional Metabolism of Antarctic Krill Product Protein in Rats

This work conducted a four-week metabolism test on rats to study the digestion and absorption characteristics of five protein-based krill products prepared from Antarctic krill as raw material.It aimed to provide theoretical support for the effective use of Antarctic krill protein and the development of novel protein resources.The results showed that the weight gain and true digestibility of the rats fed with krill meat,surimi and ordinary krill powder were significantly higher(P<0.05)than those of the rats fed with traditional casein.Compared to casein,proteins from the five Antarctic krill products were found to significantly improve the net protein utilization(P<0.05),and reduce the total cholesterol and triglycerides in the serum of rats(P<0.05).In summary,the Antarctic krill protein-based products with high nutritional values can be used as a potential novel protein resource in the food industry.

Identification of γ-aminobutyric acid transporter (GAT1)on the rat sperm

Some recent studies indicated that GABAergic system is involved in mammalian sperm acrosome reaction (AR), but direct evidence pertaining to the expression of gat1 in mammalian sperm is not yet demonstrated. In this study, we evaluated the presence of 67kDa GAT1 protein and mRNA in rat testis by Western blotting and reverse transcription-polymerase chain reaction. Meanwhile, immunohistochemical and immunofluorescent analyses also identified GAT1 protein on the elongated spermatid and sperm. These results indicated that rat testis is a novel site of gat1 expression. Further studies should be taken to explore the role of GAT1 protein on sperm acrosome reaction.

Cloning of rat sp56,the homologue of mouse sperm ZP3 receptor-sp56

Mouse sp56 is considered as one of the candidates for mouse zona pellucida 3 (mZP3) receptor. Up to date, its homologue has only been cloned from guinea pig, namely AM67. Based on the cDNA sequence of mouse sp56, we designed a pair of primer to amplify its homologue from rat testis cDNA. Using RT-PCR,two fragments of 743 bp and 938 bp were amplified. The PCR products show very high homology to mouse sp56. However, the 743 bp product completely lacks one of the seven Sushi domains of mouse sp56. Using the 743 bp product as the probe to detect the expression profile of sp56 in rat tissues, Northern blot shows that a ～2.0 kb mRNA expresses specifically in testis. Employed the RACE method, two full cDNA sequences of rat sp56 were obtained. A Mr ～42 KD band was detected in denatured and non-reducing protein sample of rat testis and sperm with anti-mouse sp56 monoclonal antibody by Western blot method. Rat sp56was localized on rat sperm head by the indirect immunofiuorescence method. Rat sp56 immunoreactivitywas detected from the early pachytene spermatocytes and throughout the spermatogenesis. Its cloning willfurther our understanding of the mechanism of the sperm-egg recognition and binding.

Protein profiles in various epididymal segments of normal and castrated rats

Aim: Epididymal proteins are known to play an important role in the maturation of spermatozoa, we ougnt to deter-mine if there are regional differences in androgen-dependent epididymal proteins. Methods: A group of adult rats wascastrated and epididymides were removed three days following castration. The epididymides were dissected into caput,corpus and cauda segments, homogenized, and proteins were fractionated by anion exchange HPLC. Proteins in select-ed fractions were resolved by SDS-PAGE and visualized by silver staining. Results: It was observed that the levels ofmultiple proteins drastically reduced in the various regions of epididymis of the orchiectomized rats. Conclusion: Theepididymal proteins appear to be useful markers to study androgenic action in the epididymis.

Effect of <i>Pleurotus ostreatus</i>on hyperglycemia, DNA damage and chromosomes aberrations

One of the main health problems with high and markedly increased complications is diabetes. Despite several projects with preventative strategies and armories of medication, the arrangement of diabetes remains grossly unsatisfactory. Thus, it is vital to identify unfamiliar drugs or novel nutraceuticals for treating and preventing diabetes without side effects. The present study deals with scientific information on mushrooms with regards to its potential use as anti-diabetic active food. In addition to the anti-hyperglycemic action of mushrooms, the present study presents its effect on DNA damage, chromosome aberrations and sperm alternations in streptozotocin-induced diabetic rats. These animals have been treated, for 30 days, with amaryl (as control treatment) (0.03 mg/kg·b·wt/dl), low-dose mushroom (100 mg/kg·b·wt/dl) and high-dose mushroom (200 mg/ kg·b·wt/dl). The glucose level GL of streptozotocin-induced diabetic animals has been markedly improved by mushroom treatment;for example GL has decreases from 167.6 mg/dl down to 116.0 mg/dl for treatment with high-dose mushroom and 128.9 mg/dl for treatment with low-dose mushroom, comparing with amaryl treatment that decreases GL down to 92.6 mg/dl. But, the experimental results show that treatment with mushroom is better than treatment with amaryl in case of genetic changes (DNA fragmentation, disappear of some base pairs and chromosome aberrations. So, it is proposed that more close scientific attention be paid to precede more research of functional mushrooms for preventive and curative treatments for diabetes.

Rat recombinant β-defensin 22 is a heparin-binding protein with antimicrobial activity

Approximately 40-50 β-defensins are predominantly expressed in the male reproductive system of mammals. This selective expression raises the question as to the roles of these molecules in innate immunity and fertility in the male reproductive tract. Rat β-defensin 22 is an epididymis-specific β-defensin expressed in segments 12-14 of the epididymis. This protein contains both β-defensin and lectin signature sequences, yet its antimicrobial activity and carbohydrate-binding ability have not been shown. We herein demonstrated the antimicrobial activity of recombinant rat β-defensin 22 against Escherichia coliand Candida albicans. Its lectinJike activity was also investigated by demonstrating its binding ability with heparin beads. This heparin-binding activity implies some potential roles for this defensin in determining the fertilisation capabilities of sperm.

Effects of Different Processed Products of Radix Codonopsis on Intestinal Flora of Rats with Spleen Deficiency

[Objectives]This study aimed to observe the effects of different processed products of Radix Codonopsis on intestinal flora in rats with spleen deficiency.[Methods]Rat models with spleen deficiency were established by bitter-cold purgation method with Radix et Rhizoma Rhei.Normal group,model group,Radix Codonopsis group,fried Radix Codonopsis group,rice-fried Radix Codonopsis group,honey-fried Radix Codonopsis group and bran-fired Radix Codonopsis group were designed.After subjecting to corresponding treatments,the changes in the quantity of intestinal microorganisms of the rats were detected.[Results]There was no significant change in the quantity of intestinal microorganisms of the rats in the normal group and model group.The rats in the rice-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased,and the quantity of Escherichia coli and Staphylococci reduced.The rats in the Radix Codonopsis group,fried Radix Codonopsis groups,honey-fried Radix Codonopsis group and bran-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased(the increases were smaller than those in the rice-fried Radix Codonopsis group),and the abundance of E.coli and Staphylococci reduced,close to normal levels.[Conclusions]Different processed products of Radix Codonopsis have obvious regulation effect on intestinal flora of rats with spleen deficiency,and the regulation effect of rice-fried Radix Codonopsis on rats with spleen deficiency is better than that of Radix Codonopsis,fried Radix Codonopsis,honey-fried Radix Codonopsis and bran-fried Radix Codonopsis.

The Role of Coconut Water on Nicotine-Induced Reproductive Dysfunction in Experimental Male Rat Model

The coconut water is the most nutritious wholesome beverage in all coconut producing countries. Folk medicine describes the use of coconut water to reduce toxic effects of nicotine and alcohol in reproductive function in men, but its direct evidence is lacking. The aim of this study was to investigate the effects of tender and mature coconut water consumption on nicotine induced changes in sperm quality, spermatogenic cell density, activity of testicular marker enzymes for spermatogenesis and testosterone level of male healthy rats. Male Sprague Dawley rats were treated with tender and mature coconut water intragastrically and nicotine subcutaneously for 15 days. The findings indicated that the coconut water supplementation improves epididymal spermatogenic cell density (p ≤ 0.05), sperm motility and morphology which were altered by nicotine. The activity of acid phosphatase and alkaline phosphatase was increased (p ≤ 0.05), which indicates that treatment with coconut water provides significant protection of germinal epithelium from nicotine toxicity. Coconut water also shows a significant increase (p ≤ 0.05) in testosterone levels in nicotine treated rats. Coconut water ameliorates the reproductive toxicity caused by nicotine due to presence on nutrients L-arginine, ascorbic acid, minerals like calcium and magnesium.

Effects of "Products of Chicken Embryo" on Growth and Sexual Development in Rats

Products of Chicken Embryo (PCE) such as Ji-Pei-Jing is a kind of food for Chinese children prepard from chicken embryo. Female rats on 21 days were administered with aqueous solutions of Ji-Pei-Jing (1. 2 %, 3 %, 12%, and 48%, respectively) by gavage up to their onsets of puberty.The rats in the control group were treated with distilled water. However, Ji-Pei-Jing treatment exerted some effects on sexual mauration in the immature female rats. Essentially, the effects showed a dose-response tendency with an inverted 'U' shape.The age of vopnal opening for gnup treated with Ji-Pei-Jing was significantly earier than that to the control. Its uterus weight/b. w. ratio also significantly increased on day 30 and at the first estrus. There were significantly increases in the adrenal weight/b. w. raio of 30-day-old rats that were treated with 3%, 12%, and 48% Ji-Pei-Jing. The rats treated with 48% Ji-Pei-Jing had significantly lesser ovary weight.b.w. radio on day 30, too. The rats treated with Ji-Pei-Jing could normally ovulate at the first estrus, and no significant differences were observed during estrous cycles.The effects of PCE on serum levels of K, P, LH in 30-day-old nds and FSH in 28-day-old ras were elevated significantly by 3 % Ji-Pei-Jing treatment. It appeare that the effects of PCE result from interaction of contained complex physiologically active substances. Steriods, especially estradio-17β,possibly Play a key role, and polpeptide hormones may also exert important effects.

Transcriptome profile analysis of the accompanying migratory parasitic wasp Aenasius bambawalei(=Aenasius arizonensis girault)(Hymenoptera:Encyrtidae):Genes related to fertilization involved at different stage of ovary development

Age-related declines in fertilization success have been reported for a wide range of species.The fertilization of parasitic wasps is closely related to egg production and sperm storage.Aenasius bambawalei(Hymenoptera:Encyridae)is a key parasitic wasp of the important invasive mealybug Phenacoccus solenopsis(Hemiptera:Pseudococcidae).The female offspring ratio of this parasitic wasp was declined with parental age in mass rearing under laboratory conditions.To investigate the molecular mechanisms involved in the reproduction of A.bambawalei,an extensive analysis of the impact of age on transcriptome profile of mated ovaries of this wasp was performed by comparing the gene expression profiles of various maternal ages:the early stage(ES),the intermediate stage(IS),and the advanced stage(AS).In total,358 differentially expressed genes were identified,with 17.60%(63 genes)of the changes associated with greater expression in fertilization.Moreover,the expression of serine protease 47 precursor,serine protease inhibitor 3/4,glucose dehydrogenase,fatty acyl-CoA reductase 1-like,major royal jelly,and acyl-CoA delta(11)desaturase-like was detected by real-time fluorescence quantitative(RT-qPCR).The results showed that fertilization related genes exhibited a stage-specific pattern.Egg production and sperm storage genes in A.bambawalei were significantly modified in the transcriptome,providing a starting point for the genetic dissection of fertilization.

Morphometric study of the testis and reproductive tract （including sperm granuloma） after vasectomy in mature rats

By utilizing the rabbit model, previous studies have found good evidence indicating that vasectomy-induced spermatogenic damage is pressure-mediated： the damage occurs when the occluded reproductive tract is unable to accommodate additional spermatozoa produced by the testis. More studies with the more commonly used rat model have shown, however, controversial results on whether and why the damage occurs. In this study, 12 mature male Sprague-Dawley rats were subjected to unilateral vasectomy： double ligation （without severing） of the vas deferens exposed via a small inguinal incision; 37 days after the operation, the testes, epididymides, vasa deferentia （juxta-epididymal segments）, and sperm granulomas （at the vasectomy site） were removed to obtain methacrylate resin-embedded sections and morphometric studies carried out with light microscopy. Marked spermatogenic damage with spermatids and spermatocytes depleted in the seminiferous epithelium in 43% of the seminiferous tubule profiles was demonstrated in 5 of the 12 testes on the vasectomized side, and the damage was associated with smaller or absent sperm granulomas; in the other 7 testes with essentially normal spermatogenesis, there was an increase （by 111% on average） in the volume of the tubule lumen, associated with larger granulomas or granulomas containing more spermatozoa. There was an overall increase （by 66%） in the thickness of the rete testis in the 12 testes; the epididymis or vas deferens showed no distension. It seems therefore that the spermatogenic damage induced by vasectomy in rats is pressure-mediated as well, and that variation in the damage depends mainly on the postoperative development of the sperm granuloma.

Impact of Hydroalcoholic Extract of Humulus Lupulus L. on Sperm Quality, Reproductive Organs and Hormones in Male Rats

Objective: To investigate the impact of Humulus Lupulus L. hydroalcoholic extract on the body weights, reproductive organs, sperm quality and hormone levels in male rats. Methods: By simple random sampling method, seventy male Sprague-Dawley rats were randomly assigned to 7 groups including control group [distilled water, 1 mL/(kg·d)], Tween 80 group [25% Tween 80 solution, 1 mL/(kg·d)], olive oil group [olive oil, 1 mL/(kg·d)], diethyl stilbestrol(DES) group [DES, 100 μg/(kg·body weight)], H50, H150 and H450 [50, 150 and 400 mg/(kg·d) of Humulus Lupulus L extract, respectively]. The administration was performed via gavage once daily for 7 weeks. Body and reproductive organs weights including testes, seminal vesicles, epididymis and prostate were weighted and epididymal sperm quality were determined by digital balance. Blood samples were collected and serum free testosterone(T), luteinizing hormone(LH), follicle-stimulating hormone(FSH), and estrogen(E2) levels were measured by rat specific enzyme-linked immunosorbent assay. Results: The percentage increase in mean body weights of rats in the DES and H50, H150 and H450 groups decreased significantly compared to olive oil and Tween 80 groups(all P<0.05). The weights of seminal vesicle, epididymis and testes in rats receiving H50 were significantly higher than the control group(P<0.05 or P<0.01). The sperm count in the rats receiving H50 was significantly lower than the control group(P<0.05). The sperm motile characteristics of the rats receiving hydroalcoholic extract and DES were significantly lower than those of the control or rats receiving vehicles(all P<0.05). In H50, H150, H450 and DES groups, T and LH levels were decreased, and E2 was significantly increased compared to the control(P<0.05 or P<0.01). The FSH level did not change in all groups(P>0.05). Conclusion: Humulus Lupulus L. extract significantly increased the seminal vesicle and testes weights and reduced the sperm motility.

PRODUCTION OF TRANSFORMING GROWTH FACTOR-β BY CULTURED RAT MESANGIAL CELLS

The purpose of this study was to examine whether transforming growth factor-β (TGFβ acts as an autocrine cytokine in cultured mesangial cells. Measangial cell conditioned media (CM) were prepared and tested for their effect on mesangial cell proliferation. CM showed a concentration dependent inhibition on mesangial cell proliferation and the activity was enhanced by treating conditioned media with acid. Gel filtration analysis showed peak inhibitory activity to reside in fractions with an estimated molecular weight range of 16-30 KD. The activity was partially blocked by anti-TGFβ antibody, but not nonimmune control IgG. The presence of TGFβ was confirmed using the mink lung epithelial cell assay. Furthermore, the addition of anti-TGFβ antibody directly into culture media significantly enhanced mesangial cell proliferation. These results demonstrate that measangial cell produce both active and latent forms of TGFβ, which functions as an autocrine growth inhibitor for mesangial cells.