Gamal A Sosa

Benha University, Banhā, Muhafazat al Qalyubiyah, Egypt

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Publications (7)7.7 Total impact

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    ABSTRACT: Vitrification is a common method which is successfully used for cryopreservation of gametes and embryosin in several animal species, though subsequent progress is still limited,especially in buffalo. The present study aimed to evaluate the effect of vitrification process on nuclear status in matured buffalo oocytes. Morphology, viability, state of nuclear maturation in meiotic stages and maturation rate (matured oocyte/survived oocyte) were assessed in vitrified matured buffalo oocytes.Immature buffalo oocytes were matured in-vitro for 22 hrs. and vitrified at room temperature (25°C) in a mixture of ethylene glycol (EG) and dimethyl sulfoxide (DMSO) with 0.25 ml straw. Vitrification solutions (VS) were VS (1.5M EG+ 1.5M DMSO) 1 and VS (3M EG+ 3M DMSO). Cryoprotectants were added in two steps, with the first step concentration half 2 that of the second (and final) step concentration. After warming, normal oocytes were cultured for further 2 hrs., stained with trypan blue for viability evaluation, then fixed and stained with orcein 1% stain for detection of meiotic stages. Oocytes that reached Telophase I (TI) or Metaphase II (MII) stages were considered matured. Results showed that morphologically normal and viable oocytes were significantly decreased in vitrified oocytes than control. The percentage of oocytes reached TI and MII in un-vitrified (control) group was significantly (P<0.05) higher than those vitrified after maturation. The maturation rate was higher in control than vitrified oocytes (76.04±2.29 vs. 66.46±2.25). In conclusion, vitrification is a successful method for cryopreservation of matured buffalo oocytes. However, inspite of higher rates of viability and maturation obtained in current study, it is still lower than un-vitrified oocytes.
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    ABSTRACT: The current study was designed to find out the effect of different doses (8, 12 or 16 µg) of GnRH analogue (Buserelin acetate, BA) on the ultrasonographic picture and biometry of buffalo bulls' reproductive organs. Animals (n= 9, aged 15-18 months) were allocated into three groups (n= 3/group) according to the hormonal dose that was administered once weekly for six weeks. Results revealed that the testicular diameter and epididymal tail width did not differ significantly before and after treatment.
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    ABSTRACT: To clarify the effect of lactation period on ovarian follicular activity and associated hormonal levels in goats, six goats were monitored daily by ultrasonographic examination with blood sampling during early (Days 5 to 25; Day 0 was the day of kidding) and late (Days 40 to 60) lactation. While the presence of a corpus luteum of pregnancy retarded follicular growth in the ipsilateral ovary until Days 11-13 postpartum, the total follicular number (TFN) and area (TFA) increased during late lactation due to the significant increase in the number of medium- and large-sized follicles and decrease in the number of small follicles. Four goats showed a similar pattern of follicular development during the period studied characterized by the emergence of five and six waves during the early and late lactation, respectively. The largest follicle diameter of the first three waves monitored during early lactation was significantly smaller as compared with the diameter of those existing during late lactation. TFN showed a positive correlation with FSH but showed a negative correlation with immunoreactive (ir-) inhibin and estradiol during the postpartum period. TFA was positively correlated with ir-inhibin, estradiol and PRL and negatively correlated with FSH during the monitored periods. The plasma levels of ir-inhibin and progesterone were significantly higher during late lactation compared with the levels recorded during early lactation. Ir-inhibin levels showed a significant positive correlation with LH and estradiol during early and late lactation but showed a negative correlation with FSH during the whole lactation period. LH was positively correlated with estradiol and PRL during early and late lactation, respectively. These results suggest that the lactation period has a detrimental effect on ovarian activity during the early postpartum period in goats.
    Journal of Reproduction and Development 10/2011; 58(1):61-8. DOI:10.1262/jrd.11-012S · 1.76 Impact Factor
  • 4th Sci Conf Al-Kasr 25-28 May 2011, Al-Kasr; 05/2011
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    ABSTRACT: Melatonin is a multifunctional molecule that mediates several circadian and seasonal reproductive processes. The exact role of melatonin in modulating reproduction, however, is not fully understood-especially its effects on the ovarian follicles and oocytes. This study was conducted to investigate the expressions of the ASMT and melatonin-receptor MTNR1A and MTNR1B genes in bovine oocytes and their cumulus cells, as well as the effects of melatonin on oocyte nuclear and cytoplasmic maturation in vitro. Cumulus-oocyte complexes (COCs) from abattoir ovaries were cultured in TCM-199 supplemented with melatonin at concentrations of 0, 10, 50, and 100 ng/ml. The expression of ASMT, MTNR1A, and MTNR1B genes was evaluated by RT-PCR. Moreover, the effects of melatonin on cumulus cell expansion, nuclear maturation, mitochondrial characteristics and COCs steroidogenesis were investigated. Furthermore, the level of reactive oxygen species (ROS) was evaluated in denuded oocytes. Our study revealed that ASMT and MTNR1A genes were expressed in COCs, while the MTNR1B gene was expressed only in oocytes. Additionally, melatonin supplementation at 10 and 50 ng/ml to in vitro maturation medium significantly enhanced oocyte nuclear maturation, cumulus cell expansion and altered the mitochondrial distribution patterns, but had no effects on oocyte mitochondrial activity and COCs steroidogenesis. Melatonin-treated oocytes had a significantly lower level of ROS than controls. The presence of melatonin receptors in COCs and its promoting effects on oocyte nuclear and cytoplasmic events, indicate the potentially important roles of this hormone in regulating bovine oocyte maturation. Moreover, the presence of ASMT transcript in COCs suggests the possible involvement of these cells in melatonin biosynthesis.
    Molecular Reproduction and Development 02/2011; 78(4):250-62. DOI:10.1002/mrd.21295 · 2.68 Impact Factor
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    ABSTRACT: The present study was conducted to characterize follicular development and its hormonal control during early pregnancy in goats. The ovaries of goats (n=8) were scanned daily for follicles (> or = 2 mm in diameter) and corpora lutea by transrectal ultrasound with blood sampling from the jugular vein for monitoring the hormonal changes during the first thirty-five days after mating. During early pregnancy, three (37.5%), four (50%) and one (12.5%) goat showed nine, eight and seven waves of follicular development, respectively. The corpora lutea were detected as early as Day 3.61 ± 0.45 (7.47 ± 0.43 mm) of pregnancy (Day 0=day of mating) and attained their maximal cross-sectional diameter (10.64 ± 0.37 mm) on Day 25.7 ± 0.8 of pregnancy, respectively. A transient rise in FSH levels was temporally associated with the day of follicular wave emergence (up to three days prior to wave emergence). The plasma LH and estradiol levels were negatively correlated with the progesterone concentration. The rise in plasma immunoreactive (ir) inhibin levels was negatively correlated with the FSH concentration and positively correlated with the number of large-sized follicles. Alternatively, the mean plasma ir-inhibin levels showed a noticeable decline with the progression of pregnancy. The present results demonstrated that follicular development during early pregnancy shows a wave-like pattern, with seven to nine waves developing until Day 35 after breeding, and that the number of follicular waves can be predicted by the number of FSH peaks. The current study also demonstrated that the role of inhibin as an FSH regulator is maintained throughout early pregnancy.
    Journal of Reproduction and Development 10/2010; 56(5):520-6. DOI:10.1262/jrd.09-179T · 1.64 Impact Factor
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    ABSTRACT: The current study was performed to follow up the circulating hormonal changes and to correlate the findings with the physiological activity of the corpus luteum (CL) and placenta during pregnancy in goats. Blood samples were collected weekly from five goats during pregnancy for measuring steroid and protein hormones. A gradual increase was observed in immunoreactive (ir-) inhibin, with maximal levels at the 17th week. The plasma concentrations of estradiol and prolactin (PRL) showed nearly similar patterns during pregnancy, where they declined to basal levels during the first 4 weeks post-breeding and then increased significantly, with the maximal concentration during late pregnancy. The plasma FSH and LH concentrations were maintained at basal levels throughout the gestation period. The plasma progesterone concentration abruptly increased in the first week post-breeding and remained at high values throughout the pregnancy period. Immunohistochemical localization of inhibin alpha, beta(A), beta(B) and steroidogenic enzymes cytochrome P450 aromatase, 3alpha-hydroxysteroid dehydrogenase (3betaHSD), cytochrome 17alpha-hydroxylase P450 and cholesterol side-chain cleavage cytochrome P450 in the cyclic and pregnant goat CL revealed positive immunoreactivity without affinity differences between the luteal and pregnancy stages. The placental syncytiotrophoblasts also showed positive staining, except for inhibin beta(A) and 3betaHSD. The giant binucleate cells of the placenta showed positive immunoreactions to PRL. These results suggest that the high concentrations of ir-inhibin, estradiol and PRL during late pregnancy are of placental origin and that the placenta may have a vital role in the maintenance of pregnancy, regulation of mammary growth and preparation for kidding and lactation in goats.
    Journal of Reproduction and Development 04/2010; 56(2):243-50. DOI:10.1262/jrd.09-159S · 1.64 Impact Factor