FSH prevents depletion of the resting follicle pool by promoting follicular number and morphology in fresh and cryopreserved primate ovarian tissues following xenografting

Background: Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting. Methods: Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles. Results: Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues. Conclusions: FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation

FSH prevents depletion of the resting follicle pool by promoting follicular number and morphology in fresh and cryopreserved primate ovarian tissues following xenografting

Background: Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting. Methods: Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles. Results: Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues. Conclusions: FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation

Early gonadotropin secretion and sexual maturation in bull calves and ram lambs

Regulation of the early rise in gonadotropin secretion in bull calves and its involvement in the reproductive development of bull calves and ram lambs were studied. Development of the testes, prostate and seminal vesicles was assessed from birth to puberty by ultrasonagraphy and image intensity analysis in bull calves and ram lambs. A GnRH superagonist ( Leuprolide acetate) was used to suppress early gonadotropin secretion in bull calves and ram lambs, to study the importance of early gonadotropin secretion for testicular development. GnRH treatment was used to induce LH secretion in the early postnatal period in bull calves and its effects on testicular and endocrine development were studied. Histology and flowcytometry were used to evaluate testicular development in these studies. The role of estradiol in the negative feedback regulation of early gonadotropin secretion was studied in bull calves by using an aromatase inhibitor (ICI 16949A) and a nonsteroidal antiestrogen (ICI 182,780). The involvement of opioidergic and dopaminergic neuronal systems in the regulation of the early rise in gonadotropin secretion was studied in bull calves by using their receptor antagonists (naloxone and pimozide, respectively). Suppressing the early increase in gonadotropin secretion stressed its importance for testicular development in bull calves. The timing of the early increase in secretion of gonadotropins is also important; by inducing higher gonadotropin secretion early in the life of bull calves, reproductive development was enhanced. In ram lambs, high gonadotropin secretion up to 11 weeks of age did not appear critical for reproductive development. We were unable demonstrate a role for estradiol in the negative feedback regulation of early gonadotropin secretion in bull calves. Opioidergic and dopaminergic neuronal systems appeared to be important in the regulation of gonadotropin secretion in bull calves towards the end of the early increase in LH secretion; opioids may be critical for termination of the early rise in LH secretion. Ultrasonography and computer assisted image intensity analysis appeared to be very useful techniques in assessment of reproductive development in bull calves and ram lambs. Image intensity analysis showed promise for identifying critical stages in development of the reproductive system

Two-dimensional and three-dimensional ultrasonography for pregnancy diagnosis and antenatal fetal development in Beetal goats

Aim: The objective of this study was to compare two-dimensional (2D) and three-dimensional (3D) study of the pregnant uterus and antenatal development of the fetus. Materials and Methods: 2D and 3D ultrasound were performed from day 20 to 120 of gestation, twice in week from day 20 to 60 and once in week from day 60 to 120 of gestation on six goats. The ultrasonographic images were obtained using Toshiba, Nemio-XG (Japan) 3D ultrasound machine. Results: On the 20th day of gestation, earliest diagnosis of pregnancy was done. First 3D ultrasonographic image of the conceptus, through transabdominal approach, was obtained on day 24. On 39th day, clear pictures of conceptus, amniotic membrane, and umbilicus were seen. On 76th day of gestation, internal organs of fetus viz heart, kidney, liver, urinary bladder, and stomach were seen both in 2D and 3D images. 3D imaging showed better details of uterine structures and internal organs of the fetus. Conclusions: Comparing 3D images with 2D images, it is concluded that 2D was better in visualizing fluid while 3D images were better to view details of attachment of fetus with endometrium

Transabdominal color doppler ultrasonography: A relevant approach for assessment of effects of uterine torsion in buffaloes

Aim: The present study was conducted on advanced pregnant buffaloes suffering from uterine torsion to assess the status of fetus and uterus by transabdominal ultrasonography, and the findings were compared with normal advanced pregnant buffaloes. Materials and Methods: The study was conducted on 20 clinical cases of uterine torsion and 20 normal advanced pregnant buffaloes (control group). The lower ventral area just lateral to linea alba (on both sides of the udder) in standing animals was scanned transabdominally by the two-dimensional convex transducer for various ultrasonographic findings. The data collected were statistically analyzed by “one-way ANOVA” and “independent sample t-test” using computerized SPSS 16.0 software program. Results: Transabdominal ultrasonography revealed dead fetus in 95% uterine torsion cases and proved useful in imaging internal structures of fetuses while no dead fetus was reported in the control group. Size of umbilicus was found significantly decreased (p0.05) in uterine torsion group. Average thickness of the uterine wall and mean pixel values of fetal fluids (echogenicity) were found significantly increased (p<0.05) in uterine torsion affected buffaloes in comparison to control group. Conclusion: Status of fetus (whether live or dead), internal status of uterus, and its contents could be determined by transabdominal ultrasonography in uterine torsion cases and thus determining the prognosis of the uterine torsion cases before going for further manipulations. This will also help in taking all the precautions to avoid death of the fetus

Doppler sonography for evaluation of hemodynamic characteristics of fetal umbilicus in Beetal goats

Aim: The objective of this study was to evaluate the hemodynamic characteristics of umbilical vessels in healthy pregnant Beetal goats. Materials and Methods: Doppler examinations were performed from day 20 to 120 of gestation, twice in week from day 20 to 60 and once in week from day 60 to 120 of gestation on six goats. Results: Free floating umbilical cord was identified on day 39 of gestation. The umbilical cord waveform was characterized by the simultaneous presence of arterial and venous flow. The pattern of blood flow in the umbilical artery was represented as saw tooth pattern above the baseline. Pattern of blood flow in umbilical vein was flat and wavy in nature; presented below the baseline. Peak systolic velocity (PSV) increased significantly from day 39 to 67 and further between 98 and 120 days of gestation (p<0.05), but there was no significant increase or decrease in end-diastolic velocity (EDV). Pulsatility index (PI) value was increased significantly during 42 to 48 days of gestation and decreased significantly from 98 to 105 days of gestation. On other days, there was no significant increase or decrease. Value of resistance index (RI) was more stable than PI values as there was no significant increase or decrease in RI value. Conclusions: From the present study, it is reasonable to conclude that the normal blood flow parameters like PI, RI, PSV and EDV during gest ation might be helpful in assessment of antenatal development of fetus in the goat. This work provides the basis for further contribution in diagnosing and monitoring high-risk pregnancy in this species