• Reproductive and Developmental Biology
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• v.37 no.2
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• pp.65-73
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• 2013

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.

• Journal of the Korean Applied Science and Technology
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• v.37 no.1
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• pp.91-101
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• 2020

The purpose of this study was to investigate the influence of obesity on the expression of autophagy-related proteins in cardiac muscle. To this end, obesity was induced in rats through 20 weeks of high-fat diet, and the animals were then subjected to 8 weeks of treadmill exercise. Subsequently, the expression of proteins that regulate the induction of autophagy, formation of autophagosome, and fusion of autophagosome and lysosome was confirmed. Obesity was induced in the experimental animals (SD rats) through 20 weeks of high-fat diet (carbohydrate: 20%, fat: 60%, and protein: 20%), and they were subsequently subjected to 8 weeks of treadmill exercise (5 days/week, 30 min/day, 5 minutes; 8m/min, 5 minutes; 11m/min, 20 minutes; 14m/min). The experimental groups comprised the normal diet control group (ND-CON, n=10), high-fat diet comparison group (HFD-CON, n=10), and high-fat exercise group (HFD-TE, n=10). Oral glucose tolerance test was conducted before and after 8 weeks of treadmill exercise, and the area under the curve (AUC) was calculated. Through fasting insulin and fasting glucose levels, HOMA-IR, which is an index of insulin resistance, and abdominal visceral fat/body weight (AVF/BW) were calculated for comparison. Moreover, autophagy-related proteins were analyzed from cardiac tissue to investigate the effects of exercise training. Obesity was successfully induced in the HFD-CON group through long-term high-fat diet, and the HFD-CON group had higher body weight, AUC, HOMA-IR, and AVF/BW compared to the ND-CON group. The HFD-TE group, which underwent 8 weeks of treadmill exercise, showed improvements in AUC, HOMA-IR, and AVF/BW. Although the body weight tended to decrease as well, there was no statistically significant difference. mTOR and AMPK, which are involved in the induction of autophagy, both decreased in obesity but increased upon exercise. Beclin-1, BNIP3, ATG-7, p62, and LC3, which are related to the formation of autophagosomes, all increased in obesity and decreased after exercise. Cathepsin L and LAMP2, which regulate the fusion of autophagosome and lysosome, both decreased in obesity and increased upon exercise. Physical activity, including treadmill exercise, was found to induce normal autophagy and improve pathological phenomena observed in metabolic diseases. Therefore, the findings suggest the need to consider treadmill exercise as a primary means to achieve effective prevention and treatment of cardiac diseases.

• Horticultural Science & Technology
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• v.34 no.1
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• pp.102-111
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• 2016

The goal of this study was to characterize the BrDSR (Drought Stress Resistance in B. rapa) gene and to identify the expression network of drought-inducible genes in Chinese cabbage under drought stress. Agrobacterium-mediated transformation was conducted using a B. rapa inbred line ('CT001') and the pSL100 vector containing the BrDSR full length CDS (438 bp open reading frame). Four transgenic plants were selected by PCR and the expression level of BrDSR was approximately 1.9-3.4-fold greater than that in the wild-type control under drought stress. Phenotypic characteristics showed that BrDSR over-expressing plants were resistant to drought stress and showed normal growth habit. To construct a co-expression network of drought-responsive genes, B. rapa 135K cDNA microarray data was analyzed to identify genes associated with BrDSR. BrDSR was directly linked to DARK INDUCIBLE 2 (DIN2, AT3G60140) and AUTOPHAGY 8H (ATG8H, AT3G06420) previously reported to be leaf senescence and autophagy-related genes in plants. Taken together, the results of this study indicated that BrDSR plays a significant role in enhancement of tolerance to drought conditions.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.269-279
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• 2013

Low efficiency of somatic cell nuclear transfer (SCNT) is attributed to incomplete reprogramming of transfered nuclei into oocytes. Trichostatin A (TSA), histone deacetylase inhibitor and 5-aza-2'deoxycytidine (5-aza-dC), DNA methylation inhibitor has been used to enhance nuclear reprogramming following SCNT. However, it was not known molecular mechanism by which TSA and 5-aza-dC improve preimplantation embryo and fetal development following SCNT. The present study investigates embryo viability and gene expression of cloned porcine preimplantation embryos in the presence and absence of TSA and 5-aza-dC as compared to embryos produced by parthenogenetic activation. Our results indicated that TSA treatment significantly improved development. However 5-aza-dC did not improve development. Presence of TSA and 5-aza-dC significantly improved total cell number, and also decreased the apoptotic and autophagic index. Three apoptotic-related genes, Bak, Bcl-xL, and Caspase 3 (Casp3), and three autophagic-related genes, ATG6, ATG8, and lysosomal-associated membrane protein 2 (LAMP2), were measured by real time RT-PCR. TSA and 5-aza-dC treatment resulted in high expression of anti-apoptotic gene Bcl-xL and low pro-apoptotic gene Bak expression compared to untreated NT embryos or parthenotes. Furthermore, LC3 protein expression was lower in NT-TSA and NT-5-aza-dC embryos than those of NT and parthenotes. In addition, TSA and 5-aza-dC treated embryos displayed a global acetylated histone H3 at lysine 9 and methylated DNA H3 at lysine 9 profile similar to the parthenogenetic blastocysts. Finally, we determined that several DNA methyltransferase genes Dnmt1, Dnmt3a and Dnmt3b. NT blastocysts showed higher levels Dnmt1 than those of the TSA and 5-aza-dC blastocysts. Dnmt3a is lower in 5-aza-dC than NT, NTTSA and parthenotes. However, Dnmt3b is higher in 5-aza-dC than NT and NTTSA. These results suggest that TSA and 5-aza-dC positively regulates nuclear reprogramming which result in modulation of apoptosis and autophagy related gene expression and then reduce apoptosis and autophagy. In addition, TSA and 5-aza-dC affects the acetylated and methylated status of the H3K9.