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Discovery of porcine maternal factors related to nuclear reprogramming and early embryo development by proteomic analysis.

Zhao Q, Guo Z, Piao S, Wang C, An T - Proteome Sci (2015)

Bottom Line: In this study, we showed most of porcine oocytes became nuclear matured at 33 h of IVM and the rate had no significant difference with oocytes at 42 h of IVM (p > 0.05).Moreover, the cleavage and blastocyst rates of SCNT and PA embryos derived from 42O were significantly higher than that of 33O (p < 0.05).These results present a unique insight into maternal factors related to nuclear reprogramming and early embryo development.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Northeast Forestry University, 26 Hexing Road, Xiangfang Dist., Harbin, Helongjiang 150040 China.

ABSTRACT

Background: Differentiated cell nuclei can be reprogrammed to a pluripotent state in several ways, including incubation with oocyte extracts, transfer into enucleated oocytes, and induced pluripotent stem cell technology. Nuclear transfer-mediated reprogramming has been proven to be the most efficient method. Maternal factors stored in oocytes have critical roles on nuclear reprogramming and early embryo development, but remain elusive.

Results: In this study, we showed most of porcine oocytes became nuclear matured at 33 h of IVM and the rate had no significant difference with oocytes at 42 h of IVM (p > 0.05). Moreover, the cleavage and blastocyst rates of SCNT and PA embryos derived from 42O were significantly higher than that of 33O (p < 0.05). But 33O could sustain IVF embryo development with higher cleavage and blastocyst rates comparing to 42O (p < 0.05). To clarify the development potential difference between 33O and 42O, 18 differentially expressed proteins were identified by proteomic analysis, and randomly selected proteins were confirmed by Western blot. Bioinformatic analysis of these proteins revealed that 33O highly synthesized proteins related to fertilization, and 42O was rich in nuclear reprogramming factors.

Conclusions: These results present a unique insight into maternal factors related to nuclear reprogramming and early embryo development.

No MeSH data available.


The porcine oocytes during IVM. a The polarbody extrusion during IVM of porcine oocytes. The polarbody extrusion mainly began at 25 h (14.45 %) of IVM, and the rate at 33 h reached the plateau. The rates of porcine oocyte polarbody extrusion at 33 h (76.72 %) and 42 h (81.80 %) of IVM had no significant difference (p > 0.05). b Fluorescence micrographs of nuclei in porcine MII oocytes (200×). DNA was blue, and the MII nuclei was marked by “*”
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Fig1: The porcine oocytes during IVM. a The polarbody extrusion during IVM of porcine oocytes. The polarbody extrusion mainly began at 25 h (14.45 %) of IVM, and the rate at 33 h reached the plateau. The rates of porcine oocyte polarbody extrusion at 33 h (76.72 %) and 42 h (81.80 %) of IVM had no significant difference (p > 0.05). b Fluorescence micrographs of nuclei in porcine MII oocytes (200×). DNA was blue, and the MII nuclei was marked by “*”

Mentions: To get the time lapse characteristics of maturation of porcine oocytes during IVM, we detected the rate of 1st polarbody extrusion from 16 h to 42 h. The oocyte began to extrude polarbody mainly at 25 h (14.45 %) of IVM and reached a plateau at 33 h (Fig. 1), and there was no significant difference between the rates at 33 h (76.72 %) and 42 h (81.80 %) of IVM (Additional file 1: Table S2; p > 0.05). We also found most of 33O (79.71 %) were arrested at the MII stage, and it had no remarkable difference with 42O (83.86 %; Table 1). Theoretically, the maturation quality of 42O must be better than 33O [17–21].Fig. 1


Discovery of porcine maternal factors related to nuclear reprogramming and early embryo development by proteomic analysis.

Zhao Q, Guo Z, Piao S, Wang C, An T - Proteome Sci (2015)

The porcine oocytes during IVM. a The polarbody extrusion during IVM of porcine oocytes. The polarbody extrusion mainly began at 25 h (14.45 %) of IVM, and the rate at 33 h reached the plateau. The rates of porcine oocyte polarbody extrusion at 33 h (76.72 %) and 42 h (81.80 %) of IVM had no significant difference (p > 0.05). b Fluorescence micrographs of nuclei in porcine MII oocytes (200×). DNA was blue, and the MII nuclei was marked by “*”
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493956&req=5

Fig1: The porcine oocytes during IVM. a The polarbody extrusion during IVM of porcine oocytes. The polarbody extrusion mainly began at 25 h (14.45 %) of IVM, and the rate at 33 h reached the plateau. The rates of porcine oocyte polarbody extrusion at 33 h (76.72 %) and 42 h (81.80 %) of IVM had no significant difference (p > 0.05). b Fluorescence micrographs of nuclei in porcine MII oocytes (200×). DNA was blue, and the MII nuclei was marked by “*”
Mentions: To get the time lapse characteristics of maturation of porcine oocytes during IVM, we detected the rate of 1st polarbody extrusion from 16 h to 42 h. The oocyte began to extrude polarbody mainly at 25 h (14.45 %) of IVM and reached a plateau at 33 h (Fig. 1), and there was no significant difference between the rates at 33 h (76.72 %) and 42 h (81.80 %) of IVM (Additional file 1: Table S2; p > 0.05). We also found most of 33O (79.71 %) were arrested at the MII stage, and it had no remarkable difference with 42O (83.86 %; Table 1). Theoretically, the maturation quality of 42O must be better than 33O [17–21].Fig. 1

Bottom Line: In this study, we showed most of porcine oocytes became nuclear matured at 33 h of IVM and the rate had no significant difference with oocytes at 42 h of IVM (p > 0.05).Moreover, the cleavage and blastocyst rates of SCNT and PA embryos derived from 42O were significantly higher than that of 33O (p < 0.05).These results present a unique insight into maternal factors related to nuclear reprogramming and early embryo development.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Northeast Forestry University, 26 Hexing Road, Xiangfang Dist., Harbin, Helongjiang 150040 China.

ABSTRACT

Background: Differentiated cell nuclei can be reprogrammed to a pluripotent state in several ways, including incubation with oocyte extracts, transfer into enucleated oocytes, and induced pluripotent stem cell technology. Nuclear transfer-mediated reprogramming has been proven to be the most efficient method. Maternal factors stored in oocytes have critical roles on nuclear reprogramming and early embryo development, but remain elusive.

Results: In this study, we showed most of porcine oocytes became nuclear matured at 33 h of IVM and the rate had no significant difference with oocytes at 42 h of IVM (p > 0.05). Moreover, the cleavage and blastocyst rates of SCNT and PA embryos derived from 42O were significantly higher than that of 33O (p < 0.05). But 33O could sustain IVF embryo development with higher cleavage and blastocyst rates comparing to 42O (p < 0.05). To clarify the development potential difference between 33O and 42O, 18 differentially expressed proteins were identified by proteomic analysis, and randomly selected proteins were confirmed by Western blot. Bioinformatic analysis of these proteins revealed that 33O highly synthesized proteins related to fertilization, and 42O was rich in nuclear reprogramming factors.

Conclusions: These results present a unique insight into maternal factors related to nuclear reprogramming and early embryo development.

No MeSH data available.