Seven rc mutant and seven normal male birds (Rhode Island Red suie, RIR) were used in this study to determine the effects of rc mutation on semen characteristics, testosterone profile and spermatogenic tissues. All birds were randomly selected at week 12 of age and housed in individual cages and were fed and watered ad libitum. The birds were exposed to a 14L:10D light cycle during experiment. Semen were collected at weeks 22 to 23 from each bird twice a week and evaluated for semen volume (SV), sperm concentration (SC), total sperm count (TSC), percent of sperm motility (%SM), dead sperm (%DS), and sperm metabolic activity (SMA). To determine the testosterone concentration (TC) in plasma, blood was collected at weeks 12, 16 and 18. Testicular tissue were collected, processed and evaluated for semineferous tubule diameter (STD), round spermatid number (RSN), percent elongated sperm (%ES) and semineferous tubules length (STL). Body weight (BW), comb weight (CW) and testes weight (TW) were weighted at the end of experiment (week 23). The SV, TSC and %SM were significantly higher in normal birds but the %DS was higher in blind birds (p<0.05). The SC did not differ significantly between the two groups but its value was higher in normal birds. The sperm metabolic activity in the first h of collection did not differ significantly between the two groups but after 24 h, the level of SMA in normal group was significantly higher (p<0.05). The level of TC did not differ significantly between the two genotype groups but normal birds had higher TC in all collections except the last one. The STD, RSN, %ES and STL in normal birds were higher when compared to blind birds but the differences were insignificant except for ES percent. The BW, CW and TW between the two groups did not differ significantly but the weights were higher in normal group compared to blind birds. Statistical analysis of semen characteristics, testosterone profile and histological factors were indicated detrimental effects of rc mutation in prepubertal RIR blind male birds due to lack of light.
Present study aimed to investigate recovering effect of propolis on blood components and tissues of mouse after low dose irradiation. It is verified that the contents of Fe, Mg, P, Zn and Cu in propolis dosed blood are increased slightly than irradiated blood, however, the contents of Ba and Pb are decreased to one tenth than irradiated blood and the contents of Fe and P are increased to 10% than control group. We consider this result as the propolis acts a role of defence factor minimizing changes of elements caused by irradiation in blood. Among the blood components, Glutamate oxaloacetate transaminase (GOT) value is increased after the radiation but after dosed with propolis and irradiated the value is decreased, suggesting that propolis as a buffering material against irradiation. After dosed with propolis, a number of spermatogenic cells are lowered in testis tissue, however, nucleus and cytoplasm are clearly observed in spermatogonia, spermatocytes and spermatid cells. And nucleus and membrane of cells in the proximal convoluted tubule of renal tissue are clearly observed. Also, cytoplasmand membrane of surface mucous cells in stomach tissue are appeared in normal which is almost like those of control group. We consider that the propolis used in this study is preventing deformations of cells increasing resistance capacity against irradiation rather than recovering damaged tissues.
To investigate the cadmium (Cd) toxicity on the testis, male rats were treated with 1, 2, 4 and 8 mg/kg of Cd by IP. According to histochemical studies, Cd-treated testis tissue showed death of spermatozoa, death of Sertoli cells, death of all the spermatogenic cells, and finally disappearance of basal lamina of seminiferous tubules with increasing doses, and showed decreased ground substances and Leydig cells, increased inflammatory cells and fibroblasts, and fibroblasts, and finally disappearance of ground substances and all the cells except fibroblasts within interstitial tissues with increasing doses. According to biochemical studies, two kinds of proteins, 25 and 45 kDa, were dramatically disappeared from the total protein of rat testis treated with Cd comparing to normal testis. The result of electrophoresis of total protein suggests that actin (45 kDa), presumed on its mmolecular weight and amount, in the testis-cells is the primary target of Cd poisoning. Although its exact mechanism is not clear, the disappearance of two proteins when testis is exposed to Cd should give some clues to understnad the mechanism of necrosis of testis tissue crumbling by heavy metal pollutant such as Cd.
Objective: These studies were undertaken to evaluate the effects of the different administration duration of Morindae Radix extract solution on spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testes and the activities of sperm hyaluronidase. Materials and Method: We used 8-week-old ICR mice and administered 0.3mg/g extract solution of Morindae Radix once a day for 30, 60, 90 and 120 days. The control group was administered normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis. We also compared the testicular tissue, especially seminiferous tubules, between the control and treated groups by histochemical methods. Finally, we observed the difference of sperm hyaluronidase activities between the control and treated groups. Results: Significant administration duration-dependent differences were observed in the concentration of total sperm, motility and normality of spermatozoa of the Morindae Radix extract solution administered groups compared to the control group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae Radix extract solution administered groups compared to the control group. Also, the activity of hyaluronidase was significantly increased in the Morindae Radix extract solution administered groups compared to the control group. Conclusions: This study shows that the beneficial effect of Morindae Radix extract solution on the concentration, motility and morphology of sperm, the testicular tissues and the activities of sperm hyaluronidase increased the greater the duration the mice were administered it. We suggest that Morindae Radix may be useful for the treatment of male sexual dysfunction and infertility.
Abstract - Global decline in wildlife mammals has been accelerated during past decades. Especially the conservation the wild life mammals in polar areas, is urgent. In an effort to understand the reproduction of the seals dwelling in the polar area, spermatogenesis in the seals was reviewed. Seals breed seasonally and in most of the seal species, delayed implantation is frequently observed. To date, histological and endocrinological evaluation revealed highly cyclic nature in supermatogenesis and steroidogenesis in testis. Seasonal changes in blood testosterone level together with melatonin is closely related with changes in light cycle between summer and winter. In adult testis at breeding seasons, spermatogenesis is manifested by consecutive 18 stages of germ cell development. Three kinds of Leydig cells different in steroidogenic activity as well as cellular morphology appear during the testis development. During non-breeding season, spermatogenic arrest and Leydig cell hypoplasia are frequently found. Interestingly, blood circulation through the anastomoses of pelvic veins cooled the testes and thus guarantees spermatogenesis within the body trunk. Endocrine disruptors and heavy metals have been found in the body tissues of several seals species and alter steroidogenesis in seals, suggesting environmental pollutants together with decrease in habitats are potentially threatening the reproductive success in seal species.
The decrease in sperm quality under heat stress causes a great loss in animal husbandry production. In order to reveal the mechanism underlying the sperm quality decrease caused by heat stress, we first established a mild heat-treated mouse model. Then, the sperm quality was identified. Further, the testicular proteome profile was mapped and compared with the control using 2D electrophoresis and mass spectrometry. Finally, the differential expressed proteins involved in the heat stress response were identified by real-time PCR and Western blotting. The results showed that heat stress caused a significant reduction in mouse sperm quality (P<0.05). Further, 52 protein spots on the 2D gel were found to differ between the heat-shocked tissues and the control. Of these spots, some repair proteins which might provide some explanation for the influence on sperm quality were found. We then focused on Bag-1, Hsp40, Hsp60 and Hsp70, which were found to be differently expressed after heat shock (P<0.05). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorders.
Mesna has been used with ifosfamide to prevent urotoxicity in the treatment of testicular cancers. This drug also protected the toxicities of adriamycin without compromising cytostatic activity. With an idea of radioprotective role of sulfhydryl group of radioprotectors and of mesna decreasing the toxic effect of adriamycin which produces free radicals, mesna and radiation were administered to mice to study the protective effect of this drug and to identify the difference in regenerative capacity of the germ cells in the testis between radiation-treated and both mesna-and radiation-treated groups. The shape and numbers of spermatogenic cells in the seminiferous tubules were examined every week after irradiation. In both groups, initial reduction and later recovery in germ cell numbers and shape was observed. The lowest germ cell number was found around three weeks after irradiation. Mean germ cell number of the mesna-treated group was significantly higher than radiation-treated group at all observed periods (p<0.05). More competent regeneration was present in mesna-treated group. These results suggest that mesna protect the testis from radiation injury. Further study will be necessary to identify whether mesna protects other tissues from radiation and it does not hamper tumor control.
Purpose : These studies were undertaken to evaluate the effects of Allii tuberosi Semen (ATS) on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Materials and Methods : We used the 8-week-old mice and administered the 0.2 ml extract solution of ATS in the different concentration (0.1 mg/ml, 1 mg/ml, 10 mg/ml and 100 mg/ml) once a day for 60 days. The control group was administered the distilled water in the same way. After the administration of each extract solution, we examined the number of total, motile and normal sperm, the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Also we observed the histological changes of isolated testis. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results : The concentration of total sperm, the motility and normality of spermatozoa were significantly increased in ATS groups, especially in 1 and 10 mg/ml groups, compared to control group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the ATS groups compared to the control group, respectively. Also, the activity of hyaluronidase was significantly increased in the ATS groups compared to the control group. In the antioxidant activity analysis, the activity of testicular peroxidase was significantly increased in the ATS groups compared to the control group, especially in 1 mg/ ml group. The activity of testicular catalase was increased in ATS groups. Conclusion : This study shows that ATS has the beneficial effect on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase and testicular peroxidase. We can suggest that ATS extract solution be useful for the treatment of male sexual dysfunctions and infertility.
Spermatogenesis is known to be regulated by a number of genes and several factors such as hormones, growth factors, cytokines and others. This study was done to evaluate the relationship between HSPs and DAZ genes in human spermatogenesis; we observed the expression pattern of HSP gene in azoospermia men with DAZ gene that regulated the gene expression related with human spermatogenesis. RT-PCR method was used to detect DAZ, HSP70A, and HSP70B transcripts in all RNA samples. Total RNA was extracted from 21 testis tissues using TRIZOL reagent. cDNAs were synthesized with reverse transcriptase, AMV. All PCR reaction were performed on a PCR themocycler with DAZ, HSP70A, and HSP70B-specific primers. Semen analysis, karyotyping and testis histology were performed. DAZ gene, known as a candidate gene of azoospermia factor(AZF), was deleted in 2 of 21 patients. To evaluate the only effects of HSPs in this patients, 2 DAZ deleted cases were removed. We observed the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered to be immature. In conclusion, HSP70B as well ad DAZ gene seem to be involved causing spermatogenic failure. We suggest that HSP70B plays an important role in spermatogenesis and it is one of factors induced sperm maturation in human.
Objective: The aim of the current study is to investigate the relationship between prohibitin (PHB), capping actin protein of muscle Z-line beta subunit (CAPZB), and tektin-2 (TEKT2) and sperm motility in Murrah buffalo. Methods: We collected the high-motility and low-motility semen samples, testis, ovary, muscle, kidney, liver, brain and pituitary from Murrah buffalo, and analysed the expression of PHB, CAPZB, and TEKT2 in mRNA (message RNA) and protein level. Results: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) result showed that the expression of PHB was higher and CAPZB, TEKT2 were specifically expressed in testis as compared to the other 6 tissues, and that in testis, the expression of TEKT2 was higher than that of CAPZB and PHB. Immunohistochemistry test revealed that all three genes were located on the convoluted seminiferous tubule and enriched in spermatogenic cells. Both qRT-PCR and Western Blot results showed that the expression levels of PHB, CAPZB, and TEKT2 were significantly lower in the low-motility semen group compared to the high-motility semen group (p<0.05). Conclusion: The expression levels of PHB, CAPZB, and TEKT2 in Murrah buffalo sperm have a high positive correlation with sperm motility. And the three genes may be potential molecular markers for the decline of buffalo sperm motility.
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