Nuclear morphology, diameter and meiotic competence of buffalo oocytes relative to follicle size

Yousaf, Muhammad; Chohan, Kazim R.

doi:10.1071/rd03006

Cited by 32 publications

(24 citation statements)

References 42 publications

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“…In the present work, the average diameter of dromedary oocyte is 166.2 ± 2.6 lm and the ooplasm is black in color. These findings indicate that dromedary oocytes are larger in size than bovine (Lechniak et al 2002) or buffalo oocytes (Yousaf and Chohan 2003). The dark appearance of the ooplasm is consistent with that described in llama (Del Campo et al 1994), and dromedary oocytes ; it may be attributed to the prevalence of large quantity of lipid droplets.…”

Section: Discussionsupporting

confidence: 82%

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Morphology of Dromedary Camel Oocytes and their Ability to Spontaneous and Chemical Parthenogenetic Activation

Abdoon

Kandil

Berisha

et al. 2007

Reprod Domestic Animals

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The present work was conducted to examine (1) the morphology of dromedary cumulus-oocytes complexes (COCs), (2) to study the incidence of spontaneous development of oocytes in vivo and (3) to assess the ability of in vitro matured dromedary oocytes to chemical parthenogenetic activation compared with in vitro fertilized (IVF) oocytes. COCs were recovered from dromedary ovaries classified according to their morphology into six categories. Oocyte diameter was measured using eye piece micrometer. For chemical activation, COCs with at least three layers of cumulus-cells were in vitro matured (IVM) in TCM 199 + 10 microg/ml FSH + 10 IU hCG/ml + 10% FCS + 50 microg/ml gentamycin. COCs were incubated for 40 h at 38.5 degrees C under 5% CO2 in humidified air. After IVM, matured oocytes with first polar body (first Pb) were divided into two groups. Group 1: activated in 7% ethanol (E) for 5 min followed by culture in 2 mM 6-dimethylaminopurin (6-DMAP, E D, subgroup 1) or 10 microg/ml cycloheximide (CHX, E CHX, subgroup 2) for 3.5 h at 38.5 degrees C under 5% CO2. In group 2, oocytes were activated using 50 microM Ca A23187 (Ca A) for 5 min followed by culture in 2 mM 6-DMAP (Ca D, subgroup 3) or 10 microg/ml CHX(Ca CHX, subgroup 4) for 3.5 h at 38.5 degrees C under 5% CO2. For control group, IVM oocytes were fertilized using frozen-thawed camel spermatozoa separated by swim-up method then suspended in Fert-TALP medium supplemented with 6 mg/ml BSA (FAF) + 10 microg/ml heparin. In all groups, oocytes were in vitro cultured in SOFaa medium + 5% FCS and 5 microg/ml insulin + 50 microg/ml gentamycin. Cleavage rate and embryo development were checked on Days 2, 5 and 8. An average of 11.3 +/- 0.3 COCs were recovered/dromedary ovary. Categories 1 and 2 represented 33.1% and 34.8%, respectively, and were significantly higher (p < 0.01) than the other categories (19.1, 9.2 and 2.6% for categories 3-5, respectively). Category 6 (embryo-like structures) represented 1.2% of the recovered oocytes, staining of these embryo-like structures with orcien dye indicated the presence of divided cells with condensed nuclei. Dromedary oocytes averaged 166.2 +/- 2.6 microm in diameter with black cytoplasm. Chemical activation of IVM dromedary oocyte with first Pb in 7% ethanol or 50 microM Ca A followed by culture in 2 mM 6-DMAP showed significantly higher (p < 0.01) cleavage and developmental rates to the morula stage than oocytes activated using 7% ethanol or 50 microM Ca A followed by 10 microg/ml CHX or in vitro fertilized control group. Higher (p < 0.01) proportion of oocytes sequentially cultured in 10 microg/ml CHX or that in vitro fertilized were arrested at the 2-4-cell stage compared with that cultured in 6-DMAP.

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Section: Discussionsupporting

confidence: 82%

“…These findings indicate that dromedary oocytes are larger in size than bovine (Lechniak et al. 2002) or buffalo oocytes (Yousaf and Chohan 2003). The dark appearance of the ooplasm is consistent with that described in llama (Del Campo et al.…”

Section: Discussionmentioning

confidence: 82%

Morphology of Dromedary Camel Oocytes and their Ability to Spontaneous and Chemical Parthenogenetic Activation

Abdoon

Kandil

Berisha

et al. 2007

Reprod Domestic Animals

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“…In accordance with the results of Raghu et al, (2002) and Amer et al, 2008), granulosa cells play an essential role in maturation and blastocyst stage development in vitro (Tanghe et al, 2002). In addition, buffalo oocytes recovered from follicles ranged from (4-<6 and 6-<8mm) reached the M-II (67.1% and 79.1% respectively) than (2-<3 and 3-<4mm) (Yousaf and Chohan, 2003). However, oocytes from follicles (>4mm) were revealed to having good results in IVF in buffalo (Yousaf and Chohan, 2003. In contrast, increased fertilization, cleavage and embryo development were significantly higher in COCs more than 145 microm which can be aspirated from large follicles (> 8 mm) followed by medium and small-sized normal follicles (Raghu et al, 2002).…”

Section: Discussionsupporting

confidence: 84%

Effect of Season and Ovarian Morphology of Egyptian Buffalo on Oocyte Quality and In Vitro Maturation Rate

Mansor

2019

Journal of Current Veterinary Research

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The aim of the current study was to investigate the effect of season, follicle size and corpus luteum (CL) on yield, quality and in vitro maturation of Egyptian buffalo oocytes. Ovaries were collected from abattoirs all over the year and cumulus oocytes complexes (COCs) were aspirated from visible small (≤3 mm), medium (3-6 mm) and large (>6 mm) follicles of normal ovaries. Ovaries were classified according to bearing mature CL3 into with and without CL. Oocytes quality were classified into good, fair, poor and denuded. Only good and fair quality oocytes were used for in vitro maturation in TCM-199 medium supplemented with 10% FCS, 10 µg/ml ovine FSH, 10 IU/ml hCG, 1 µg/ml Estradiol and 1% antibiotic antimycotic mix. Results revealed that a significant difference between the four seasons with significant increase of oocyte yield, quality and maturation for winter and spring than autumn and summer seasons. Oocytes aspirated from medium sized follicles (3-6 mm) were of higher in quality and maturation rate than large (> 6 mm) and small size follicles (≤ 3 mm). Recovery rate, quality and maturation rate of oocytes were higher for ovaries without CL than ovaries with CL. In conclusion, the winter season, medium sized follicles and ovaries without CL showed positive effects on recovery rate, quality and in vitro maturation of buffalo oocytes.

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“…The results for IVF in buffalo are very poor as compared to cattle because majority of IVF studies in buffalo use the protocols which are established for cattle (Yousaf and Chohan, 2003). The overall success rate of embryo production following IVF in buffalo is reported to be not more than 10-15 percent which is very low when compared to that of cattle (Chauhan et al, 1998;Nandi et al, 2001;Raghu et al, 2002).…”

Section: Introductionmentioning

confidence: 93%

Real-Time PCR Based Expression Study of Cyclin B Gene in Buffalo Cumulus Oocyte

Yadav¹

2014

Adv. Anim. Vet. Sci.

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The success rate of in vitro fertilization (IVF) in buffalo is very low, though a research of more than a decade has been carried out. One of the reasons could be the lack of standardised protocol for in vitro maturation of oocytes. The present study was conducted to determine the temporal expression of Cyclin B gene in follicles of different size. A good quality cumulus oocyte complexes (COCs) were recovered from large (>6 mm) and medium (2-6 mm) follicles of buffalo ovaries by aspiration. The recovered COCs were matured in Tissue Culture Medium-199 medium supplemented with 10% foetal bovine serum (FBS), 0.4% bovine serum albumin (BSA) and hormones pregnant mare's serum gonadotropin (10 IU/mL) and human chorionic gonadotropins (10 IU/mL) and incubated for zero min, 30 min, 60 min, 90 min, 18 hr, 20 hr, 22 hr, 24 hr and 26 hr in a CO 2 incubator with 5% CO 2 under humid conditions at 38.5°C. Total RNA was isolated from COCs, reverse transcribed and real time PCR was done to study the expression pattern of Cyclin B gene in follicles of different sizes. Real time PCR assay revealed higher expression of Cyclin B in medium size follicles. However temporal expression of Cyclin B was highest at zero min in both large and medium follicles and decreases as the maturation time increases. These results suggest that the role of Cyclin B is more during early phases of maturation and less during later stages of maturation. All copyrights reserved to Nexus® academic publishers

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Nuclear morphology, diameter and meiotic competence of buffalo oocytes relative to follicle size

Cited by 32 publications

References 42 publications

Morphology of Dromedary Camel Oocytes and their Ability to Spontaneous and Chemical Parthenogenetic Activation

Morphology of Dromedary Camel Oocytes and their Ability to Spontaneous and Chemical Parthenogenetic Activation

Effect of Season and Ovarian Morphology of Egyptian Buffalo on Oocyte Quality and In Vitro Maturation Rate

Real-Time PCR Based Expression Study of Cyclin B Gene in Buffalo Cumulus Oocyte

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