scholarly journals Growth Arrest Specific Gene 6 Improved in vitro Maturation of Oocytes and the Development Potential of Porcine Embryo after Parthenogenetic Activation

2021 ◽  
Author(s):  
Zhi-Peng Li
Keyword(s):  
In Vitro Maturation ◽  
Growth Arrest ◽  
Specific Gene ◽  
Development Potential ◽  
Parthenogenetic Activation ◽  
Porcine Embryo

47 EFFECT OF L-CARNITINE TREATMENT DURING OOCYTE MATURATION ON THE POST-THAW DEVELOPMENT OF PORCINE EMBRYOS VITRIFIED AT THE PRONUCLEAR STAGE

2016 ◽  
Vol 28 (2) ◽  
pp. 153 ◽  
Author(s):  
C. G. Grupen ◽  
T. Somfai ◽  
K. Kikuchi
Keyword(s):  
Lipid Droplets ◽  
In Vitro Maturation ◽  
Survival Rates ◽  
Cell Number ◽  
Blastocyst Formation ◽  
Porcine Embryo ◽  
Porcine Oocyte ◽  
Proportional Data ◽  
Bovine Oocytes

The extreme cryo-sensitivity of porcine oocytes and embryos is attributed to their endemically high content of cytoplasmic lipid droplets. In attempts to improve the cryo-tolerance of porcine embryos, various strategies have been used to reduce the amount of lipid droplets present in the cytoplasm before vitrification. Recently, the cryo-tolerance of bovine oocytes vitrified at the metaphase II stage was improved by supplementing in vitro maturation (IVM) medium with l-carnitine (LC), a stimulator of lipid metabolism (Chakitisakul et al. 2013 Theriogenology 79, 590–598). The objective of this study was to determine the effect of supplementing IVM medium with LC on the post-thaw development of porcine embryos vitrified at the pronuclear stage. Oocytes recovered from the ovaries of prepubertal gilts were matured in modified porcine oocyte medium supplemented with 0 (control) or 12 mM LC during the final 22 h of IVM. Following IVF, presumptive zygotes were cultured in porcine zygote medium-3. At the pronuclear stage, cohorts of embryos from each group were either vitrified using a solid surface vitrification procedure (Somfai et al. 2009 Biol. Reprod. 80, 42–49) or cultured for 7 d without being vitrified. Vitrified zygotes were subsequently warmed and cultured for 7 d. The rates of cleavage, blastocyst formation, and hatching were recorded, and all blastocysts were stained to determine the total cell numbers. Three replicates were performed. Proportional data were arcsine transformed and subjected to ANOVA, and cell number data were analysed by t-test. The post-thaw survival rates of the embryos that were vitrified did not differ between the groups (control: 95.7%; LC: 97.1%; P > 0.05). There were no significant effects of LC treatment or vitrification on the rates of cleavage, blastocyst formation, and hatching (Table 1). Vitrified embryos derived from LC-treated oocytes produced blastocysts with fewer cells than vitrified embryos derived from untreated oocytes (Table 1; P < 0.05). In contrast to previous findings in other species, the results indicate that supplementing IVM medium with LC did not enhance the post-thaw development of porcine embryos vitrified at the pronuclear stage. Table 1.Effect of l-carnitine (LC) treatment and vitrification on porcine embryo development C. Grupen was supported by an OECD Fellowship.

scholarly journals Integrin alpha 5 beta 1 expression negatively regulates cell growth: reversal by attachment to fibronectin.

1995 ◽  
Vol 6 (6) ◽  
pp. 725-740 ◽  
Author(s):  
J A Varner ◽  
D A Emerson ◽  
R L Juliano
Keyword(s):  
Cellular Proliferation ◽  
De Novo ◽  
Cell Attachment ◽  
Growth Arrest ◽  
Early Gene ◽  
Immediate Early ◽  
Specific Gene ◽  
Cellular Quiescence

Cells selected for overexpression of the integrin alpha 5 beta 1 show decreased proliferation and loss of the transformed phenotype. We provide evidence that de novo expression of the integrin alpha 5 beta 1 in HT29 colon carcinoma cells results in the growth arrest of these cells as characterized by reduced DNA synthesis and cellular proliferation in vitro. In fact, expression of integrin alpha 5 beta 1 on these cells induces the transcription of growth arrest specific gene 1 (gas-1), a gene product known to induce cellular quiescence, but blocks transcription of the immediate early genes c-fos, c-jun, and jun B. In vivo, the alpha 5 beta 1 transfectants display dramatically reduced tumorigenicity as well as a highly differentiated phenotype when compared with their pSVneo-transfected counterparts. Surprisingly, ligation of alpha 5 beta 1 on these cells by cell attachment to a fibronectin substrate not only reverses the growth inhibition and gas-1 gene induction but activates immediate early gene transcription. These findings demonstrate that integrin alpha 5 beta 1 expression in the absence of attachment to fibronectin activates a signaling pathway leading to decreased cellular proliferation and that ligation of this receptor with fibronectin reverses this signal, thereby contributing to the proliferation of transformed cells.

Relationship between follicle size and oocyte developmental competence in prepubertal and adult pigs

2007 ◽  
Vol 19 (7) ◽  
pp. 797 ◽  
Author(s):  
Melanie A. Bagg ◽  
Mark B. Nottle ◽  
David T. Armstrong ◽  
Christopher G. Grupen
Keyword(s):  
In Vitro Maturation ◽  
Fold Increase ◽  
Cell Number ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Parthenogenetic Activation ◽  
Follicle Size ◽  
Low Efficiency ◽  
The Relationship

The present study compared the distribution and steroid composition of 3-, 4- and 5–8-mm follicles on the surface of prepubertal and adult ovaries, and determined the relationship between follicle size and developmental competence of oocytes following parthenogenetic activation. The effect of 1 mm dibutyryl cAMP (dbcAMP) for the first 22 h of in vitro maturation (IVM) on the embryo development of prepubertal oocytes from the three follicle size cohorts was also determined. Compared with adult, prepubertal ovaries contained a higher proportion of 3-mm follicles (46 v. 72%, respectively), but a lower proportion of 4-mm (33 v. 22%, respectively) and 5–8-mm follicles (21 v. 6%, respectively). Adult follicular fluid (FF) contained 11-fold higher levels of progesterone (P4) than prepubertal FF, with similar levels observed between all adult follicle sizes. In prepubertal FF, the P4 concentration increased with follicle size from 3 to 4 to 5–8 mm. Rates of blastocyst development following parthenogenetic activation of adult oocytes from all three follicles sizes were similar (approximately 55%), whereas rates from prepubertal oocytes increased with increasing follicle size from 3 (17%) to 4 (36%) to 5–8 mm (55%). Treatment with dbcAMP for the first 22 h of IVM led to a 1.5-fold increase in the rate of blastocyst development for prepubertal oocytes from 3-mm follicles, but had no effect on prepubertal oocytes from the 4 and 5–8 mm classes. Mean blastocyst cell number increased with follicle size in prepubertal ovaries and was similar for all follicle sizes in adult ovaries. The present study demonstrates that the low efficiency of in vitro embryo production observed using prepubertal compared with adult pig oocytes is due to a greater proportion of 3-mm follicles on prepubertal ovaries, which contain oocytes of inferior developmental competence.

163 Treatment of Growth Differentiation Factor 8 (GDF8) During Porcine Oocyte In Vitro Maturation Changed Transcriptional Pattern and AKT Signaling

2018 ◽  
Vol 30 (1) ◽  
pp. 221
Author(s):  
J. D. Yoon ◽  
E. Lee ◽  
S.-H. Hyun
Keyword(s):  
Protein Expression ◽  
Western Blotting ◽  
Gene Transcription ◽  
In Vitro Maturation ◽  
Specific Gene ◽  
Pcr Array ◽  
Maternal Factors ◽  
Realtime Pcr ◽  
Differentiation Factor

Growth differentiation factor 8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. The purpose of this study was to investigate the effects of GDF8 on porcine oocytes during in vitro maturation (IVM). We investigated specific gene transcription levels in oocytes and cumulus cells (CC) after IVM by realtime PCR array, and specific protein expression and activation levels in matured CC by Western blotting. To collect IVM oocytes, the cumulus–oocyte complexes (COC) were aspirated and matured in 500 μL of M199 containing 10% porcine follicular fluid and eCG and hCG for 22 h, and then cultured in M199 without hormones for 22 h. Each concentration (0, 1, 10, and 100 ng mL−1) of GDF8 was included in M199 during whole process of in vitro maturation. Data were analysed by ANOVA followed by Duncan using SPSS (IBM/SPSS, Armonk, NY, USA). Data are presented as means, and differences were considered significant at P < 0.05. After 44 h of IVM, oocytes were mechanically denuded from CC with 0.1% of hyaluronidase, and then the separated oocytes and CC were sampled following each group. To assess the effect of GDF8 on specific gene transcription level changes as a dose response during IVM, a realtime PCR array was performed. In CC, the 1 and 10 ng mL−1 GDF8 supplement groups showed that transcription levels of transcription co-factors CBP and SP1, cell metabolic regulator MAPK1, and cumulus expansion-related genes Has2, Cox-2, Ptx3, and Areg were significantly different from control when hierarchically clustered by euclidean distance with average linkage method after IVM. In matured oocytes, the 10 and 100 ng mL−1 GDF8 supplement groups showed the maternal factors JMJD3 and Zar1, transcriptional regulator FOXO1, Sirt1, and Sirt2, mitochondrial activity factor Sirt3, ACSL3, and ACADL, anti-apoptosis gene BCL-2, and oocyte secrete factor BMP15 mRNA transcription levels were significantly different compared with control. To determine the effect of GDF8 supplement during IVM, the oocyte maturational regulator AKT protein expression and phosphorylation levels analysed in CC by Western blotting. The 10 and 100 ng mL−1 supplement groups showed significantly increase phosphorylated (P)-AKT per total (T)-AKT (1.22 and 1.18 times higher than control) protein levels (P < 0.05). In conclusion, supplement of GDF8 during IVM activates FOXO homologue transcription and induces cumulus cell expansion via activation of AKT signalling in CC. During the process of IVM, the changes in transcriptional landscape in CC may result in accumulation of maternal factors and mitochondrial activation in oocytes. This work was supported, in part, by a grant from the ‘the Next-Generation BioGreen 21 Program (Project No. PJ011288, PJ011077)’ Rural Development Administration and the ‘National Research Foundation of Korea Grant funded by the Korean Government (NRF-2016R1D1A1B03933191, NRF-2017R1A2B4002546)’, Republic of Korea.

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