Ultrastructural and morphometrical changes of mice ovaries following experimentally induced copper poisoning.

BACKGROUND: Copper (Cu) is an essential trace element involved in normal reproduction but its overexposure may produce some detrimental effects. The aim of this study was to investigate the effects of copper sulfate poisoning on morphometery of mice ovarian structures and probable intracellular changes. METHODS: Thirty mature female mice were randomly allocated to control and two treatment groups. In treatment groups, two different doses of copper sulfate including 100 mg/kg and 200 mg/kg in 0.2 cc were applied once a day for 35 consecutive days by gavage. Control animals received normal saline using the same volume and similar method. Animals from each experimental group were sacrificed 14 and 35 days after the beginning of drug administration and the left ovaries were removed for stereological evaluations by light microscopy and right ovaries were obtained for preparing electron microscopic sections. RESULTS: The morphometrical results showed that only the number of antral follicles was decreased by 100 mg/kg copper sulfate on day 14 compared to the control group (P=0.043). Hence, higher copper dose or longer consumption period significantly reduced different classes of follicles and corpora lutea. With 100 mg/kg copper sulfate some mild ultrastructural cell damages such as decrease of zona pellucida thickness, limited vacuolated areas and nuclear envelop dilation were seen on day 14. Higher or longer Cu administration produced more detrimental effects including more vacuolated areas, presence of secondary lysosomes, irregularity in cell shape and segmented nuclei with condensed and marginated chromatin and more enlarged and damaged mitochondria. CONCLUSION: New evidences of early as well as late intracellular damages of copper has been presented by accurate stereological and ultrastructural methods. Antral follicles was the most susceptible cells with the lower and shorter copper consumption and long term or higher dose of copper affected the whole of ovarian structures.

Evaluation of epididymal sperm quality following experimentally induced copper poisoning in male rats.

The trace element copper has been identified as a highly toxic element for spermatozoa. Our goal in this study was to assess relationship between copper poisoning and semen quality parameters. In this study, sixty male Wistar albino rats weighing 200-240 g (3.5-4 months old) were divided into three different groups of twenty rats as follows: the first group (Group 100), which was treated by gavage with copper sulphate at a dose of 100 mg kg(-1) day(-1) for 8 weeks; the second group (Group 200), which received 200 mg kg(-1) day(-1) copper sulphate by gavage during experimental period (56 days); and the control group (Group C), which received the same volume of distilled water by gavage during experimental period. The blood, semen and histopathological samples were obtained from five cases of 20 animals of each group every 2 weeks at 2, 4, 6 and 8th week. Results showed that sperm concentration, motility and viability in group 100 and 200 were significantly decreased (P < 0.05) in comparison with group C. A significant decrease in sperm concentration, motility and viability indicates the possibility of adverse effect of copper poisoning on male fertility. Copper might be mediator of the effect of oxidative damage and play an essential role in spermatogenesis and male infertility.

A study of the uterine protein variations during the estrous cycle in the cow: molecular weights determination.

This study was carried out to investigate molecular weights (MW) of the uterine protein content as the first step in their identification, and their variations during the estrous cycle as a part of the uterine adjustments for provision of an appropriate environment for physiological events in the bovine. Of the 24 pro-estrous, 21 estrous, 24 met-estrous and 46 diestrous uterine fluid samples prepared for our previous work, five samples in each phase were selected randomly and subjected to SDS-PAGE analysis in parallel with standard marker proteins to separate their protein content into fractions. By comparing these fractions with those of the standards, MWs of <14.4, 20, 30, 38, 40, 46, 67, 75, 85, 90, 160, 200, 210, 270 and 330 kiloDaltons (kDa) were calculated, respectively. Proteins with MWs of less than 14.4kDa [5.3 mg/ml (7.5% of total protein)] were observed only in met-estrus. Proteins with the MW of 20 kDa [14.0 mg/ml (19.4%)] were found only in diestrus. Proteins with MW of 30 kDa in met-estrus [20.6 mg/ml (28.9%)] and diestrus [4.5 mg/ml (6.3%)], and 38 kDa proteins were observed in a considerable amount in pro-estrus [41.4 mg/ml (44.5%)] and estrus [40.0 mg/ml (44.2%)]. Forty kiloDaltons proteins were observed only in diestrus [5.5 mg/ml (7.6%)], and 46 kDa proteins were observed only in met-estrus [5.3mg/ml (7.6%)]. Sixty seven, 75 and 330 kDa proteins were observed in all the phases of the cycle with different densities, the higher values recorded in diestrus, pro- and met-estrus, respectively. Proteins with MW of 85 kDa were observed in all the phases but diestrus [9.8 mg/ml (10.5%), 8.6 mg/ml (9.5%) and 7.3 mg/ml (10.2%) for pro-estrus, estrus and met-estrus, respectively], and proteins with MW of 90 kDa were observed in estrus [20.0mg/ml (22.1%)] and diestrus [3.3mg/ml (4.6%)]. Proteins of 160 kDa fraction were observed in pro-estrus [9.8 mg/ml (10.5%)] and met-estrus [10.7 mg/ml (15%)], 200 kDa proteins were observed only in diestrus [6.3mg/ml (8.7%)]; finally, 210 and 270 kDa proteins were observed only in estrus [3.4 mg/ml (3.8%) and 2.0 mg/ml (2.2%), respectively]. It was concluded that protein profile of the bovine uterine lumen changes during the estrous cycle, both in quality and quantity, which may be a part of uterine adaptation for the physiological events. Further identification of these proteins would give us a better insight into their role in the bovine reproduction.

A study of the uterine protein variations during the estrus cycle in the cow: a comparison with the serum proteins.

To investigate uterine protein changes during the estrus cycle in the bovine, 115 pluriparous genital tracts and blood samples were collected from the abattoir in Urmia. Genital tracts were considered healthy based on gross examination of the uterus and uterine histopathological findings. The phase of the estrus cycle was determined by the examination of the structures present on the ovaries and the uterine tonicity. Of the collected samples, 24 were pro-estrus, 21 estrus, 24 met-estrus and 46 diestrus. The uterus was incised and uterine fluid was collected by gentle scraping of the uterine mucosa with a curette. The total protein concentration, protein profiles (on agarose gel electrophoresis) in the uterine fluid were evaluated and compared with those of the serum. Total protein, alpha1, alpha2, beta1 and beta2 globulin values in the uterus were significantly higher than those of the serum (P<0.05), while, the albumin, gamma1 and gamma2 globulin values in the serum were higher than those of the uterus throughout the cycle. During pro-estrus, uterine fluid beta2 (1.96 g/dl) and serum gamma1 (1.07 g/dl) and gamma2 (1.27 g/dl) globulins were higher than those in the other phases of the cycle. During estrus, serum total protein was lower than the other phases (4.92 g/dl), which was considered to be due to a reduction in serum alpha1 (0.25 g/dl), gamma1 (0.65 g/dl) and gamma2 (0.64 g/dl) globulins in this phase. In met-estrus uterine fluid beta1 globulin was in the lowest (1.19 g/dl) and serum gamma2 globulin at a high level (1.24 g/dl). It was concluded that uterine proteins as well as serum proteins fluctuate during the estrus cycle and, except for the albumin and gamma globulins, its protein content is higher than the serum. During the follicular phase of the cycle uterine alpha globulins are higher than those in other phases, with an elevation in beta1 and a reduction in beta2 and gamma globulin values during estrus, which may reflect the preparation of the uterus for receiving spermatozoa for the conception in this phase.

Experimental reconstruction of teat mucosa by vestibular mucosal graft in cows. A histopathologic and radiographic study.

Injuries to the teat in dairy cows can result in partial or complete obstruction of the teat lumen. Different treatment techniques have been used to restore normal function in injured teats, one of which is autogenous mucosal grafts. The purpose of this study was to evaluate the vestibular mucosa as a replacement for teat mucosa in severe teat injuries. Sixteen teats of four healthy, mature, non-gravid Jersey cows were randomly divided into two equal groups. Under high epidural analgesia and after surgical preparation a 1 x 1.5 cm piece of teat mucosa was removed. In group 1, the defect was replaced by a 2 x 2.5 cm vestibular mucosa, whereas in group 2, the defect was left open. In both groups, a sterile disposable teat cannula was inserted into the teat cistern following surgery. To evaluate luminal diameter, double contrast radiography with constant air pressure was performed every 25 days till day 125, after which the animals were slaughtered and teats removed for histopathological study (H&E staining). On the basis of radiographic examination, luminal narrowing in group 2 was significantly more severe than in group 1. Histopathologically, the entire mucosal grafts of group 1 were taken and a good adhesion could be seen between the graft and the host epithelium. In group 2, severe submucosal fibrosis and mucosal papilloid hyperplasia resulted in severe narrowing of the teat cistern. According to the results of this study, it can be concluded that using vestibular mucosal grafts with temporary insertion of teat cannula can be considered as a method of treating teat mucosal injuries.