Isolation and Identification of Transforming Growth Factor β from In Vitro Matured Cumulus Oocyte Complexes

Widjiati, Widjiati; Boediono, Arief; Sumitro, Sutiman Bambang; Hinting, Aucky; Aulaniam,; Susilowati, Trinil

doi:10.4308/hjb.19.1.6

Cited by 3 publications

(3 citation statements)

References 21 publications

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“…5). The increase in TGFβ expression enhances oocyte development, steroidogenesis, and aromatization by cumulus granulosa cells, and upregulate essential transcripts required for oocyte meiosis [27, 39,40,41]. Paradoxically, RA significantly reduced TAGLN and ACTA2 expression in oocytes and cumulus cells.…”

Section: Discussionmentioning

confidence: 99%

Effects of all-trans retinoic acid on the in vitro maturation of camel (Camelus dromedarius) cumulus-oocyte complexes

et al. 2019

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All- trans retinoic acid (RA) is a metabolite of vitamin A and has pleiotropic actions on many different biological processes, including cell growth and differentiation, and is involved in different aspects of fertility and developmental biology. In the current study, we investigated the effects of RA on camel ( Camelus dromedarius ) cumulus-oocyte complex in vitro maturation (IVM). IVM medium was supplemented with 0, 10, 20, and 40 µM RA. Application of 20 µM RA significantly reduced the proportion of degenerated oocytes and significantly improved oocyte meiosis and first polar body extrusion compared to the control and other experimental groups. Retinoic acid significantly reduced the mRNA transcript levels of apoptosis-related genes, including BAX and P53 , and reduced the BAX/BCL2 ratio. In addition, RA significantly reduced the expression of the Transforming growth factor beta (TGFβ) pathway-related transcripts associated with the actin cytoskeleton, ACTA2 and TAGLN ; however, RA increased TGFβ expression in cumulus cells. The small molecule SB-431542 inhibits the TGFβ pathway by inhibiting the activity of activin receptor-like kinases (ALK-4, ALK-5, and ALK-7); however, combined supplementation with RA during IVM compensated for the inhibitory effect of SB-431542 on cumulus expansion, oocyte meiosis I, and first polar body extrusion in activated oocytes. The current study shows the beneficial effects of RA on camel oocyte IVM and provides a model to study the multifunctional mechanisms involved in cumulus expansion and oocyte meiosis, particularly those involved in the TGFβ pathway.

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Section: Discussionmentioning

confidence: 99%

Effects of all-trans retinoic acid on the in vitro maturation of camel (Camelus dromedarius) cumulus-oocyte complexes

et al. 2019

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“…Collected using a 10 ml disposable syringe with an 18 G needle previously filled with MEM media. The COC that had been collected was washed with MEM media (Widjiati et al 2012). From 20 eligible ovaries with good ovarian condition, 34 COCs met the criteria as sample unit (the immunocytochemistry process obtained 17 COCs and CLSM as many as 17 COCs).…”

Section: Cumulus-oocyte Complex Collectionmentioning

confidence: 99%

Polish Journal of Veterinary Sciences

Widjiati

Faizah

Darsini

et al. 2022

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The process of vitrification of the cumulus-oocyte complex (COCs) often results in cold shock. When warming, heat shock occurs which can disrupt the balance of intracellular calcium (Ca 2+ ) intensity. Drastic changes in temperature cause Reactive Oxygen Species (ROS), affecting changes on Ca 2+ in COCs. The role of calcium is needed for oocyte activation in the fertilization process. The purpose of this study was to measure the expression of Ca 2+ and the intensity of Ca 2+ in COCs after vitrification. The study was divided into 2 groups, the control group (C) of fresh COCs, and the treatment group (T) of COCs after vitrification. After vitrification for 24 hours, then thawing, the expression of Ca 2+ was examined using the Immunocytochemistry (ICC) method and the intensity of calcium (Ca 2+ ) with a Confocal Laser Scanning Microscope (CLSM). The research data obtained were analyzed statistically by T-Test. The results showed that the expression of Ca 2+ in the control group (12.00±0.00) was different from the treatment group (0.35±0.79). The intensity of Ca 2+ in the control group (1059.43±489.59) was different from the treatment group (568.21±84.31). The conclusion of this study is that cryopreservation affects calcium in COCs; there were differences in the expression and the intensity of Ca 2+ between fresh COCs and COCs after vitrification. Ca 2+ intensity of COCs after vitrification was concentrated in the nucleus, while in fresh COCs it was concentrated in the cytoplasm.

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“…Hasil yang sama didapatkan Widjiati et al (2012) yang mengidentifikasi TGF  pada OOK sapi setelah dimaturasi in vitro selama 20 jam. Transforming Growth Factor Beta (TGF ) juga diidentifikasi pada oosit babi yang dilakukan maturasi in vitro yang menunjukkan bahwa TGF  berperan penting dalam proses maturasi oosit (Budna et al (2017).…”

Section: Penelitianunclassified

EXPRESSION OF TRANSFORMING GROWTH FACTOR-beta AND GROWTH DIFFERENTIATION FACTOR-9 ON SHEEP OOCYTES VITRIFIED AFTER AND BEFORE IN VITRO MATURATION

Faizah

Aswin

2021

jveteriner

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Oocyte vitrification today became a hope to preserve fertility. Its was a major challenge because of oocyte characteristic in every phase. Immature oocytes were more sensitive to osmotic stress and the membrane wes less stable while mature oocyte have spindles that were very susceptible to temperature decrease. The study aim to compare the effect of vitrification before and after in vitro maturation to the expression TGF beta and GDF9. Oocyte of ewes divided into control groups (K0), K1 maturation prior vitrification, K2 vitrification prior maturation. Vitrification begins with washing oocytes in PBS supplemented of 20%serum for 1-2 minutes, followed by equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing was processed by in the media: 1). PBS + 20% serum + 0.5 M sucrose, 2).PBS + 20% serum + 0.25 M sucrose, and 3).PBS + 20% serum + 0.1 M sucrose. Immunocytochemical stain was performed to evaluate TGF ? and GDF9 expression. Remmele scale index (IRS) was used to read the result. TGF beta expression both in oocyte and cummulus of K0 and K1 was significant statistically difference (p<0.05) compare with K2. GDF9 expression both in oocyte and cummulus of K0 and K1 was significant statistically difference (p<0.05) compare with K2. We concluded that immature oocyte give better expression of TGF â and GDF9 than mature oocyte.

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Isolation and Identification of Transforming Growth Factor β from In Vitro Matured Cumulus Oocyte Complexes

Cited by 3 publications

References 21 publications

Effects of all<i>-trans</i> retinoic acid on the <i>in vitro</i> maturation of camel (<i>Camelus dromedarius</i>) cumulus-oocyte complexes

Effects of all<i>-trans</i> retinoic acid on the <i>in vitro</i> maturation of camel (<i>Camelus dromedarius</i>) cumulus-oocyte complexes

Polish Journal of Veterinary Sciences

EXPRESSION OF TRANSFORMING GROWTH FACTOR-beta AND GROWTH DIFFERENTIATION FACTOR-9 ON SHEEP OOCYTES VITRIFIED AFTER AND BEFORE IN VITRO MATURATION

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