[Effect of calligonum comosum on ovarian histology of polycystic ovary mouse model]

Background: Increased oxidative stress in women with polycystic ovary syndrome attracted attentions to antioxidant treatments. Calligonum Comosum is a planet with antioxidant propertis

Objective: This study aimed to investigating Calligonum effect on polycystic ovarian histology of polycystic ovary mouse model

Methods: Thity two female NMRI mice with 25-30 gr weight and 8 weeks age were investigated. A single dose of estradiol valerate [40 mg/kg; im] was used for induce polycystic ovarian morphology. Calligonum Comosom extract [20 mg/kg/ week; ip] was injected for 4 consequent weeks. In sham group, only DMSO was used. After 4 weeks, blood and histological samples were prepared to study

Results: There was no significant effect of 20 mg/kg Calligonum on polycystic ovarian morphology mouse model

Conclusion: The effect of other doses of Calligonum Comosum on fertility or adverse effect of this planet need to be more investigated

Isolation of bovine spermatogonial cells and co-culture with prepubertal sertoli cells in the presence of colony stimulating factor-1

Spermatogonial stem cells [SSCs] are infrequent self-renewing cells among the type A spermatogonia within the seminiferous tubules and are the basis of spermatogenesis in mammalian testis. An adequate number of SSCs is a primary requirement for the study of their behavior, regulation, and further biomanipulation. In this paper, we studied the development of the primary co-cultures of type A spermatogonia and prepubertal bovine sertoli cells in the presence of Colony Stimulating Factor 1 [CSF1], a potential contributor in the SSC niche. The effect of different concentrations of CSF1 [0, 10, 50 and 100 ng/mL] on the colonization activity of spermatogonial cells was assessed 4, 7 and 11 days after the beginning of the culture by counting the total number of colonies and measuring their area in each group of the present experiment. Immunofluorescent staining against OCT4 and vimentin led to the confirmation of the nature of both the SSCs and sertoli cells. Results showed that the total number of colonies from day 4 to 11 increased significantly in all groups, independent of CSF1 concentration. In addition, the total number and total area of colonies were higher [not significant] in 10 and 50 ng/mL CSF1 treatments than the control and 100 ng/mL CSF1 groups in all the three evaluations during the experiment. However, this difference was only significant [p<0.05] between the total area of colonies in the control and 10 ng/mLCSF1 groups at day 4 of co-culture. It was concluded that CSF1 can be a suitable growth factor for improving SSCs colonization in vitro, particularly during the first days of culture where accompanying sertoli cells still have not proliferated sufficiently to support the propagating spermatogonial cells

Effect of testosterone on spermatogonial cell colony formation during In vitro co-culture

The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells have provided a very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules consist of spermatogonial stem cells. To investigate and biomanipulate these cells, first the proliferation and viability rate of cells should be increased in vitro. In the present study, the in vitro effects of testosterone on spermatogonial cell colony formation were investigated. Sertoli and spermatogonial cells were isolated from 3-5-month-old calves. The identity of the cells was confirmed through analysis of immunocytochemistry. Co-cultured Sertoli and spermatogonial cells were treated with testosterone in different doses of 0.2 micromol L-1, 0.4 micromol L-1 and 0.8 micromol L-1, before colony assay. testosterone did not decrease the proliferation of spermatogonial stem cells.Testosterone can be chosen for in vitro colonization of spermatogonial cells with other factors

[ effects of Selenium on changes in sperm antioxidant capacity in old and adult rats]

Selenium as an antioxidant is essential for normal function of testis and spermatogenesis. It can reduce formation of free oxygen radicals and as a result it is expected to improve male fertility. The aim of this study was to evaluate alterations in antioxidant capacity of old rats sperms after prescription of 0.2 mg/kg of Selenium. In this study, 15 old male rats of 10-12 months of age and 15 adult male rats of 2-3 months of age were randomly divided into three groups: control, sham and experimental groups. Control group did not receive any treatment; sham group rats received intra peritoneal injections of equal volume of Selenium solvents [normal saline] as Selenium in experimental group. Experimental groups of male rats received daily intraperitoneal injection of Selenium [0.2 mg/kg] for 5 weeks. After 42 days from initiation of injection, the rats were killed by cervical dislocation and after obtaining sperm, total antioxidant capacity of the sperms was measured by FRAP assay. The absorbance of TPTZ-fe[+2] was read at 593 nm by spectrophotometery. For the statistical analysis, SPSS software was used and data analysis was performed by means of Kruskal-Wallis and Mann-Withney U tests. P<0.05 was considered significant. Results of this study showed significant differences in mean values of total antioxidant capacity in both old and adult rats in experimental and control groups [P<0.05]. Also comparison of mean values of antioxidant capacity of sperm solution in adult and old control groups showed a significant difference [742.26 +/- 1.06 vs. 672.061 +/- 0.78 respectively] [P<0.05]. The results of this study showed that Selenium treatment in old rats [0.2 mg/kg after 35 days] can improve total antioxidant capacity of the sperms of old rats. Regarding low levels of antioxidants in old rats, it can be suggested oxidative stress can result in diminution of antioxidant levels. Therefore antioxidant therapy could be considered as a method for improvement of the quality of sperms of old men

Assessment of piwil2 gene expression pattern upon germ cell development from mouse embryonic stem cell

In order to improve culture conditions for optimal spermatogenesis, quantitative assessment of the male germ cell gene expression profile upon spontaneous ES cell differentiation is necessary. In this study, the quantitative expression profile of Piwil2, germ-line specific marker, during the early stage of embryoid body [EB] formation and differentiation [0-3-day-old EB] was studied. CCE mouse embryonic stem cells [ESCs] were cultured in DMED containing 20% fetal bovine serum [FBS] for 1, 2 and 3 days. The total RNA was isolated from ESCs, 1-3-day-old EBs, and adult testis as positive control. cDNA was prepared and quantitative real-time PCR was done for Oct-4 to study the pluripotency of this cell line. Also, the molecular pattern of PiwiI2 expression in developing EB was investigated. Our quantitative results confirmed the pluripotency of CCE mouse ESC line and showed that PiwiI2 was expressed in undifferentiated CCE mouse ESC line. Our results also showed that expression of Piwil2 increased significantly during the process of EB formation and differentiation up to 2-day-old EB and decreased non- significantly in 3-day-old EB. Our results suggest that EB provide a cellular environment similar to the early embryonic microenvironment and cause the efficient and progressive germ cell lineage differentiation in this system

arrangement of microtubules in embryos derived from mice young, old and reconstructed oocytes

To study the structure and distribution of microtubules in embryos derived from young, old and reconstructed oocytes. Embryos obtained from old [50 embryos], young [50 embryos] and reconstructed oocytes [10 embryos] were studied by immunocytochemistry. The microtubule structures of the embryos were studied by using fluroscent microscopy with FITC-PI filter and polyclonal antibody against alfa tubulin. The spindle structure of MII young oocyte and the obtained embryos were normal with the suitable condensation. There was no contact between chromosome and spindle in old Oocytes as well as the obtained embryos, in addition, the spindle was extended in old group. In reconstructed embryos, thin and scattered filaments were observed. This study reveals that the arrangement of microtubules in reconstructed embryos was caused by repeating of injection and oocyte manipulation. Also, interactions between karyoblast, cytoplasm and microtubuls may not be suitable. This may be caused by low fertilization in these oocytes

[Effect of antioxidants supplementation on human sperm parameters after freezing]

Antioxidant reduces oxidative stress during cryo-preservation. The aim of this study was to find out the effects of vitamin E and C on sperm parameters after cryo-preservation. Human semen samples were obtained from Vali-e-asr Hospital. The samples were divided in two groups [normal and oligospermia groups]. Semen was pooled in liquid nitrogen after thawing, samples were centrifuged, then vitamin E and C were added to medium and the aliquots were incubated for 45 minutes in incubator Co2. In control group, no antioxidant was added to medium. Sperm parameters were analyzed according to WHO criteria. Data was analyzed by ANNOVA test. There was a significant increase in the progressive motility and viability of sperm which was supplemented by vitamin E, with 1, 2 Mm [p<0.05] in the normal groups [the increase in the oligospermia group, after addition of vitamin E with 1, 2, Mm was not significant]. Vitamin C did not have a significant effect on sperm parameters with 1, 2 Mm concentration. Supplementation of media with alpha-tocopherol is beneficial for sperm motility and viability rates after cryopreservation and it may be of clinical value in assisted conception procedures

Effect of EGF on the development of 16-18 celled mouse embryos and expression of EGFR after vitrification

Successful development and implantation of cryo-preserved embryos depend on suitable culture medium, cytokines, growth factors and genomic expression. Epidermal growth factor and its receptor have important roles on embryonic development and implantation. The purpose of this study was to investigate the influence of EGF on preimplantation development and to detect mRNA for the EGF receptor in vitrified mouse embryo. 8 to 16 celled mouse embryos were collected from super-ovulated NMRI mice 56 to 64 h after HCG injection and divided into 4 groups: two control groups were cultured in MEM-alpha [control 1] and MEM-alpha+EGF [10 ng/ml] as control 2. The experimental groups were vitrified immediately after collection and then cultured for 96h in MEM-alpha and M-alpha+EGF [10 ng/ml] respectively. Expression of EGFR in hatched and hatching blastocysts was studied too. Following RT, PCR nested primers were designed to optimize the specificity. The results showed that blastocyst formation and hatching rates were significantly lower in vitrified embryos and degeneration rate was significantly higher in these groups. Blastocyst formation, hatching and degeneration rates were not significantly different in control group 1 compared to control group 2 and also in exp. group 1 in comparison with exp. group 2. 96 hours after culture, mRNA expression of EGFR was detected in embryos in the four groups. In conclusion addition of EGF [10 ng/ml] to a complex medium like [MEM-alpha] doesn't have any stimulatory effect on embryonic development of vitrified and non-vitrified embryos. Vitrification did not prevent EGFR expression after 96h of culturing, but it seems that addition of EGF to the medium stimulates EGFR expression in mouse blastocysts

effects of cold injury on sensorimotor cortex of mouse brain

Cold injury has been used as a useful model for studies of traumatic brain injury. This model is used to induce brain edema. Brain edema is a pathophysiological condition of increased brain water content due to a variety of coexisting brain injuries, including ischemia, trauma, tumor and infection. The purpose of this study was to evaluate the effects of cold injury on sensorimotor cortex of mouse. 15 male NMRI mice, 6-8 weeks old every one of them with the weight of 30-35gr [5 mice per group] were studied. To produce cold injury a metal probe cooled with liquid nitrogen and was applied to the surface of the intact skull above the parietal lobe by force of 100 gr for 30 sec. Brains were removed 72h after cold injury, 10 micro m serial sections were obtained following fixation, processing and blocking of the brain. The sections were stained using creysl fast violet. Sensorimotor cortex was recognized and 35 fields were chosen randomly to be studied. To do morphometrical study on cortex of frontal and parietal lobes, the cells with a nucleus diameter of 10 micro m were determined. The data were analyzed by means of ANOVA and TUKEY'S HSD test. The results indicated that the number of alive neurons in the model group was significantly [p<0.05] lower than that of control groups. Cold injury decreases the number of normal cells of sensory motor cortex and leads to cell death

Evaluation of the efficiency of pIRES2 EGFP and pcDNA3-hBDNF-v5 plasmids in transfection of CCE ES cells by the electroporation method

Unlimited self renewal and potential capacity of embryonic stem cells [ESCs] in differentiating into a wide variety of cell types has made the cells an attractive source of donor cells for developmental studies and cell therapy. The aim of the present study was the evaluation of the transfection efficiency of pIRES2-EGFP and pcDNA3-hBDNF-v5 plasmids in CCE ES cells by the electroporation method. The plasmids transformed into DH5

[ effects of association of ultrasound with testosterone treatment on sperm parameters and mouse testis structure]

Introduction: GnRH agonists and antagonists can affect testicular function which testosterone is one of them. In spite of the broad use of ultrasound in medicine, its potential to induce cell surface changes is questionable. In the present study, administration of testosterone in association with ultrasound was studied

Materials and methods: The adult male balb/c mice were divided into the following groups: 1] control: injection of distilled water with the same volume as testosterone. 2] exp. 1: treatment with 2 5 mg/mouse testosterone. 3] exp. 2: exposure to 0.5 W/cm[2] for 2 min. 4] exp. 3: association of treatment with testosterone [25 mg/mouse] with exposure to 0.5 W/cm[2] for 2 min. The experimental period was six weeks and after finishing the experiments, the mice were killed by cervical dislocation, left testis and epididymis were removed. Sperm count, survival and motility rates were assessed. The structure of testis was evaluated after preparation. Morphometric data was analyzed using ANOVA and post hoc [TU KEY] tests

Results: The results showed that treatment with testosterone could induce azospermia and the number of sperm, viability and motility rates in the exp.1 reduced significantly comparing with the control group [P<0.05]. Ultrasound irradiation didn't have significant effects on sperm parameters and testicular structure. Association of testosterone treatment with ultrasound irradiation caused some changes as exp.1 group, but there were significant differences with exp.2 [which received exposure to 0.5 W/cm[2] ultrasound for 2 min.]

Conclusions: It is concluded that the effects of ultrasound were mechanical and thermal such as compression of tubules and its association with testosterone treatment didn't show synergism effects

effects of association of ultrasound with testosterone treatment on sperm parameters and mouse testis structure

GnRH agonists and antagonists can affect testicular function which testosterone is one of them. In spite of the broad use of ultrasound in medicine, its potential to induce cell surface changes is questionable. In the present study, administration of testosterone in association with ultrasound was studied. The adult male balb/c mice were divided into the following groups: 1] control: injection of distilled water with the same volume as testosterone. 2] exp. 1: treatment with 25 mg/mouse testosterone. 3] exp. 2: exposure to 0.5 W/cm [2] for 2 min. 4] exp. 3: association of treatment with testosterone [25 mg/mouse] with exposure to 0.5 W/cm [2] for 2 min. The experimental period was six weeks and after finishing the experiments, the mice were killed by cervical dislocation, left testis and epididymis were removed. Sperm count survival and motility rates were assessed. The structure of testis was evaluated after preparation. Morphometric data was analyzed using ANOVA and post hoc [TUKEY] tests. The results showed that treatment with testosterone could induce azospermia and the number of sperm, viability and motility rates in the exp.1 reduced significantly comparing with the control group [P<0.05]. Ultrasound irradiation didn't have significant effects on sperm parameters and testicular structure. Association of testosterone treatment with ultrasound irradiation caused some changes as exp.1 group, but there were significant differences with exp. 2 [which received exposure to 0.5 W/cm [2] ultrasound for 2 min]. It is concluded that the effects of ultrasound were mechanical and thermal such as compression of tubules and its association with testosterone treatment didn't show synergism effects