Hyperglycemia Influences Apoptosis and Autophagy in Porcine Parthenotes Developing In Vitro

서지정보

· 발행기관 : 한국동물번식학회
· 수록지 정보 : Reproductive & developmental biology / 37권 / 2호
· 저자명 : Yong-Nan Xu, Ying-Hua Li, Sung Hyun Lee, Jung-Woo Kwon, Seul Ki Lee, Young-Tae Heo, Xiang-Shun Cui, Nam-Hyung Kim

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
ocyte Collection and In Vitro Maturation Embryo Culture
Terminal Deoxynucleotidyl Transferase-Mediated dUTPNick-End Labeling (TUNEL)
AssaTerminal Deoxynucleotidyl Transferase-Mediated dUTP Nick-End Labeling (TUNEL)
Assayy
Mitochondrial Staining and Image Analysis
Immunofluorescence Staining
Real Time Reverse Transcription Polymerase Chain Reaction(Real Time RT-PCR)
Statistical Analysis
RESULTS
Effect of Different Concentrations of Glucose on PorcineEmbryos Developing In Vitro
Glucose Concentration Affects Cell Number and Apoptosisin The Blastocyst Stage
Hyperglycemia Affects Mitochondrial Distribution but not Nuclear DNA-Encoded Mitochondrial Gene Expressionin Blastocysts
Effect of Hyperglycemia on Autophagy and Autophagy-Related Gene Expression in Blastocysts
DISCUSSION
REFERENCES

영어초록

The objective of this study was to examine the effects of high concentrations of glucose on porcine parthenotes developing in vitro. Addition of 55 mM glucose to the culture medium of embryos at the four-cell-stage significantly inhibited blastocyst formation, resulting in fewer cells in blastocyst-stage embryos and increased levels of apoptosis and autophagy compared to control. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression of pro-apoptotic genes (Caspase 3, Bax and Bak) and autophagy genes (Atg6 and Atg8/Lc3) were increased significantly by the addition of 55 mM glucose to the culture medium compared to control. MitoTracker Green fluorescence revealed a decrease in the overall mitochondrial mass compared to control. However, the addition of 55 mM glucose had no effect on mRNA expression of the nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1. These results suggest that hyperglycemia reduced the mitochondrial content of porcine embryos developing in vitro and that this may hinder embryonic development to the blastocyst stage and embryo quality by increasing apoptosis and autophagy in these embryos.

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