Kinetics of GDF9 expression in buffalo oocytes during in vitro maturation and their associated development ability

Jain, Trapti; Jain, Asit; Kumar, Parveen; Goswami, S. L.; De, Sachinandan; Singh, Dheer; Datta, Tirtha Kumar

doi:10.1016/j.ygcen.2012.07.001

Cited by 9 publications

(7 citation statements)

References 55 publications

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“…Further, assessment of maturation potential of T4 group oocytes in terms of the real time expression analysis of genes involved also supported the interpretations as above. GDF9, an important transcript involved in oocyte maturation and overall development kinetics [Gode et al, 2011] has been shown to follow a normal bell shaped curve with maximum expression during mid-maturation phase [Jain et al, 2012] following which it declines. Both the groups followed the same pattern with T4 group registering significantly high expression at 8-16 h IVM interval.…”

Section: Discussionmentioning

confidence: 99%

“…Primer sequences for RPS18 and GDF9 were used as shown by Jain et al [2012] and for EIF1A, PAP, and U2AF as per Verma et al (2012). Working primer concentration optimization and reaction setup was done as described by Jain et al, 2012. PCR conditions used were 95°C for 10 min, then 40 cycles consisting of denaturation at 95°C for 15 s, annealing at 59°C for 15 s and extension at 72°C for 20 s. Control reactions were also setup simultaneously.…”

Section: Relative Quantification Of Developmentally Important Genesmentioning

confidence: 99%

“…RPS 18 transcripts were amplified as internal control. Primer sequences for RPS18 and GDF9 were used as shown by Jain et al [2012] and for EIF1A, PAP, and U2AF as per Verma et al (2012). Working primer concentration optimization and reaction setup was done as described by Jain et al, 2012.…”

Section: Relative Quantification Of Developmentally Important Genesmentioning

confidence: 99%

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Transient Arrest of Germinal Vesicle Breakdown Improved In Vitro Development Potential of Buffalo (Bubalus Bubalis) Oocytes

Kumar

Dholpuria

et al. 2017

J of Cellular Biochemistry

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Germinal vesicle breakdown (GVBD) is the first milestone that an oocyte needs to achieve toward completing the maturation and gaining potential to fertilize. Significantly lower in vitro embryo production rate in buffaloes can be attributed to heterogeneity of GVBD occurrence among oocytes obtained from abattoir derived ovaries. Evidence from our earlier work had suggested that different qualities of buffalo oocytes differ significantly in their timing of GVBD. Besides, these oocytes also differ in terms of volume of Akt phosphorylation, which initiates the process of GVBD. With objective of synchronizing the oocytes for GVBD, immature buffalo oocytes were subjected to a two-step culture protocol, initially in the presence of GVBD inhibitors and subsequently, in vitro maturation (IVM) with added SC79 (activates Akt). Expression of developmentally important genes was assessed along with embryo development rate and blastocyst health to interpret the consequences. Oocytes subjected to a short GVBD inhibition period of 6 h followed by IVM with SC79 resulted in improved cleavage and blastocyst rates. Resultant blastocysts also possessed higher ICM: TE ratio. Further, GVBD inhibited oocytes displayed a sustained cytoplasmic maturation status in terms of reorganization of cortical granules (CGs), mitochondrial membrane potential, and glutathione levels during the period of inhibition. We conclude that a temporary GVBD arrest of buffalo oocytes and modulation of Akt improves the in vitro embryo development rate as well as quality of resultant embryos. Besides, our meiotic arrest protocol does not affect the cytoplasmic maturation.

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Section: Discussionmentioning

confidence: 99%

Section: Relative Quantification Of Developmentally Important Genesmentioning

confidence: 99%

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Transient Arrest of Germinal Vesicle Breakdown Improved In Vitro Development Potential of Buffalo (Bubalus Bubalis) Oocytes

Kumar

Dholpuria

et al. 2017

J of Cellular Biochemistry

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“…The matured COCs were washed thrice in BO medium and placed in BO medium droplets. The frozen thawed buffalo semen was processed for in vitro capacitation as per the procedure described earlier by Jain et al [12] and 50 µl of the sperm suspension (at nal concentration of 1 × 10 6 cells/ml) was added to each fertilization drops having 15 COCs and incubated at 38.5 °C with 5% CO2 for 12 h. The presumptive zygotes were removed from the fertilization drops after 12 h of insemination, adhered cumulus cells were mechanically removed by vortexing and washed ve times in modi ed Charles and Rosenkrans 2 amino acid (mCR2aa) medium. Following washing, 15 presumptive zygotes were co-cultured with monolayers of granulosa cells in 100 µl drops of IVC-I medium (mCR2aa supplemented with 0.8% (w/v) BSA, 1 mM glucose, 0.33 mM pyruvate, 1 mM glutamine, 1 x MEM essential amino acid, 1x non-essential amino acid and 50 µg/ml gentamycin).…”

Section: Ivm Of Oocytes Ivf and Ivc For Production Of Early Embryosmentioning

confidence: 99%

Transient Heat-Shock Disrupts Molecular Signals and Competence of Riverine Bubalus Bubalis Oocytes and Early Embryos in a Stage-Specific Manner

Singh

Kumar

et al. 2020

Preprint

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Background Thermal stress elicits detrimental effect in reproduction performance of Bubalus bubalis, buffalo. Riverine buffalo oocytes and early embryos possess differential ability to grow and survive under thermal stress condition. It is interesting to know how precisely the short and transient heat stress impacts growth and development of buffalo oocytes and early embryos in a stage specific manner. In this study, we aim to identify the most sensitive and vulnerable stage of oocytes and early embryos against transient heat stress as well as unravel the underlying molecular signals responsible for the stunted embryo development under thermal stress. It was assessed by utilizing six different groups of oocytes and embryos with an incorporation of 4 h transient heat shock at 40ºC during different stage of in-vitro maturation (IVM) and in vitro culture (IVC). Results The stressed oocytes of group (grp) 1, 2 and 3 exhibit comparable paces of attaining metaphase-II (M-II) phase at different time interval of IVM. The most detrimental effect of heat stress was observed in grp 2 with sharp reduction in morula to blastocysts transition rate (p<0.05). Expression of mRNA transcripts of HSP8, MnSOD and Sirt-3 genes were significantly increased from mid-IVM to 4-cell embryo although subsequently, down-regulated in embryos of 8-16 cells, morulae and blastocysts. Expression of maternal to embryo transition (MET) genes viz. PAP, U2Af and eIF4A was significantly down regulated from 2-cell embryo to morula (p<0.05). Maternal recognition of pregnancy (MRP) and morulae to blastocyst transition genes were poorly expressed in grp 2 than of the other stress groups (p<0.05). Conclusions Disruption of molecular signal has implicated in the poor formation of inner cell mass (ICM) and trophectoderm (TE) cells, this results in compromised development prospects of early buffalo embryos in a stage specific manner. Well-coordinated molecular signals associated with heat stress held responsible for reduced development of early embryo. We establish the most vulnerable stage of buffalo oocytes and presumptive zygotes against transient heat shock and observe a narrow window as the most critical stage for regulating the development potential of the prospective embryos under short heat stress environment.

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“…However, it is well known that normal mammalian oocyte maturation and development require somatic cumulus cell expansion (CCE) (Jain et al 2012;Barberi et al 2013;Caixeta et al 2013;Gharibi et al 2013;Mito et al 2013). This process is regulated by, among others, paracrine signalling of oocyte-cumulus cell secreted growth factors (Su et al 2009;Romaguera et al 2010;Hussein et al 2011).…”

Section: Tgfb Superfamily Gene Expression During Oogenesismentioning

confidence: 99%

The role of TGF superfamily gene expression in the regulation of folliculogenesis and oogenesis in mammals: a review

Piotrowska¹,

Kempisty²,

Sosinska³

et al. 2013

Vet. Med.

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ABSTRACT:The normal differentiation of follicles from the preantral to the antral stage is regulated by the synthesis and secretion of several important growth factors. Moreover, the proper growth and development of the oocyte and its surrounding somatic granulosa-cumulus cells is accomplished through the activation of paracrine pathways that form a specific cross-talk between the gamete and somatic cells. It has been shown that several growth factors produced by the ovary are responsible for the proper growth and development of follicles. The developmental competence of mammalian oocytes (also termed developmental potency) is defined as the ability of female gametes to reach maturation (the MII stage) and achieve successful monospermic fertilisation. Proper oocyte development during folliculo-and oogenesis also plays a critical role in normal zygote and blastocyst formation, as well as implantation and the birth of healthy offspring. Several molecular markers have been used to determine the developmental potency both of oocytes and follicles. The most important markers include transforming growth factor beta superfamily genes (TGFB), and the genes in this family have been found to play a crucial role in oocyte differentiation during oogenesis and folliculogenesis. In the present review, we summarise several molecular aspects concerning the assessment of mammalian oocyte developmental competence. In addition, we present the molecular mechanisms which activate important growth factors within the TGFB superfamily that have been shown to regulate not only follicle development but also oocyte maturation.

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Kinetics of GDF9 expression in buffalo oocytes during in vitro maturation and their associated development ability

Cited by 9 publications

References 55 publications

Transient Arrest of Germinal Vesicle Breakdown Improved In Vitro Development Potential of Buffalo (Bubalus Bubalis) Oocytes

Transient Arrest of Germinal Vesicle Breakdown Improved In Vitro Development Potential of Buffalo (Bubalus Bubalis) Oocytes

Transient Heat-Shock Disrupts Molecular Signals and Competence of Riverine Bubalus Bubalis Oocytes and Early Embryos in a Stage-Specific Manner

The role of TGF superfamily gene expression in the regulation of folliculogenesis and oogenesis in mammals: a review

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