Effects of alpha-linolenic acid and essential amino acids on the proliferation and differentiation of C2C12 myoblasts

Zhou, Dongjie; Li, Xiao‐Han; Lee, Song‐Hee; Heo, Geun; Cui, Xiang-Shun

doi:10.12750/jarb.37.1.17

Cited by 7 publications

(5 citation statements)

References 23 publications

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“…Approximately 100 embryos in the four-cell stage from each group were collected after 48 h of culture. Western blot analysis was performed as previously reported [33]. Sodium dodecyl sulphate sample buffer (1×) was used to lyse the oocytes at 90 • C for 10 min.…”

Section: Western Blot Analysismentioning

confidence: 99%

ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos

Li,

Sun,

Jiang

et al. 2023

IJMS

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Zinc finger and SCAN domain-containing 4 (ZSCAN4), a DNA-binding protein, maintains telomere length and plays a key role in critical aspects of mouse embryonic stem cells, including maintaining genomic stability and defying cellular senescence. However, the effect of ZSCAN4 in porcine parthenogenetic embryos remains unclear. To investigate the function of ZSCAN4 and the underlying mechanism in porcine embryo development, ZSCAN4 was knocked down via dsRNA injection in the one-cell stage. ZSCAN4 was highly expressed in the four- and five- to eight-cell stages in porcine embryos. The percentage of four-cell stage embryos, five- to eight-cell stage embryos, and blastocysts was lower in the ZSCAN4 knockdown group than in the control group. Notably, depletion of ZSCAN4 induced the protein expression of DNMT1 and 5-Methylcytosine (5mC, a methylated form of the DNA base cytosine) in the four-cell stage. The H3K27ac level and ZGA genes expression decreased following ZSCAN4 knockdown. Furthermore, ZSCAN4 knockdown led to DNA damage and shortened telomere compared with the control. Additionally, DNMT1-dsRNA was injected to reduce DNA hypermethylation in ZSCAN4 knockdown embryos. DNMT1 knockdown rescued telomere shortening and developmental defects caused by ZSCAN4 knockdown. In conclusion, ZSCAN4 is involved in the regulation of transcriptional activity and is essential for maintaining telomere length by regulating DNMT1 expression in porcine ZGA.

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Section: Western Blot Analysismentioning

confidence: 99%

ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos

Li,

Sun,

Jiang

et al. 2023

IJMS

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“…A total of 60 porcine embryos per group were placed in 1× sodium dodecyl sulfate (SDS) sample buffer and heated at 95°C for 10 min. According to previous studies (D. Zhou et al, 2020, 2022). the proteins were separated using SDS‐polyacrylamide gel electrophoresis (PAGE) and transferred to polyvinylidene fluoride membranes in 1× transfer buffer.…”

Section: Methodsmentioning

confidence: 95%

“…According to previous studies (D. Zhou et al, 2020Zhou et al, , 2022. the proteins were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to polyvinylidene fluoride membranes in 1× transfer buffer.…”

Section: Western Blot Analysismentioning

confidence: 99%

Y‐box binding protein 1 influences zygotic genome activation by regulating N6‐methyladenosine in porcine embryos

Jiang

Sun

et al. 2023

Journal Cellular Physiology

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Y-box binding protein 1 (YBX1) is a member of the family of DNA-and RNA-binding proteins that play crucial roles in multiple aspects, including RNA stabilization, translational repression, and transcriptional regulation; however, its roles in embryo development remain less known. In this study, to investigate the function of YBX1 and its mechanism of action in porcine embryo development, YBX1 was knocked down by microinjecting YBX1 siRNA at the one-cell stage. YBX1 is located in the cytoplasm during embryonic development. The mRNA level of YBX1 was increased from the fourcell stage to the blastocyst stage but was significantly decreased in YBX1 knockdown embryos compared with the control. Moreover, the percentage of blastocysts was decreased following YBX1 knockdown compared with the control. Defecting YBX1 expression increased maternal gene mRNA expression and decreased zygotic genome activation (ZGA) gene mRNA expression and histone modification owing to decreased levels of N6-methyladenosine (m6A) writer N6-adenosine-methyltransferase 70 kDa subunit (METTL3) and reader insulin-like growth factor 2 mRNA-binding protein (IGF2BP1). In addition, IGF2BP1 knockdown showed that YBX1 regulated the ZGA process through m6A modification. In conclusion, YBX1 is essential for early embryo development because it regulates the ZGA process.

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“…We collected at least 70 embryos per group in 20 µL of ice-cold Laemmli sample buffer (sodium dodecyl sulfate (SDS) sample buffer containing 2-mercaptoethanol) heated at 95 °C for 10 min and stored at –20 °C. Based on previous study ( Zhou et al, 2022 ), proteins were separated by molecular weight using SDS-polyacrylamide gel electrophoresis (SDS-PAGE), then transferred to polyvinylidene fluoride membranes. The membranes were blocked in Tris-buffered saline (TBS) containing 0.1% Tween 20 and 5% skim milk for 1 h at room temperature, then incubated overnight at 4 °C with ATF6 (1:1 000; Cat # 24169-1-AP, Proteintech, China) or GAPDH (1:1 000, 5174S; Cell Signaling Technology, USA) antibodies.…”

Section: Methodsmentioning

confidence: 99%

ATF6 aggravates apoptosis in early porcine embryonic development by regulating organelle homeostasis under high-temperature conditions

Sun¹,

Jiang²,

Li³

et al. 2023

Zoological Research

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Activating transcription factor 6 (ATF6), one of the three sensor proteins in the endoplasmic reticulum (ER), is an important regulator of ER stress-induced apoptosis. ATF6 resides in the ER and, upon activation, is translocated to the Golgi apparatus, where it is cleaved by site-1 protease (S1P) to generate an amino-terminal cytoplasmic fragment. Although recent studies have made progress in elucidating the regulatory mechanisms of ATF6, its function during early porcine embryonic development under high-temperature (HT) stress remains unclear. In this study, zygotes were divided into four groups: control, HT, HT+ATF6 knockdown, and HT+PF (S1P inhibitor). Results showed that HT exposure induced ER stress, which increased ATF6 protein expression and led to a decrease in the blastocyst rate. Next, ATF6 expression was knocked down in HT embryos under microinjection of ATF6 double-stranded RNA (dsRNA). Results revealed that ATF6 knockdown (ATF6-KD) attenuated the increased expression of CHOP, an ER stress marker, and Ca 2+ release induced by HT. In addition, ATF6-KD alleviated homeostasis dysregulation among organelles caused by HT-induced ER stress, and further reduced Golgi apparatus and mitochondrial dysfunction in HT embryos. AIFM2 is an important downstream effector of ATF6. Results showed that ATF6-KD reduced the occurrence of AIFM2-mediated embryonic apoptosis at HT. Taken together, our findings suggest that ATF6 is a crucial mediator of apoptosis during early porcine embryonic development, resulting from HT-induced ER stress and disruption of organelle homeostasis.

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Effects of alpha-linolenic acid and essential amino acids on the proliferation and differentiation of C2C12 myoblasts

Cited by 7 publications

References 23 publications

ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos

ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos

Y‐box binding protein 1 influences zygotic genome activation by regulating N6‐methyladenosine in porcine embryos

ATF6 aggravates apoptosis in early porcine embryonic development by regulating organelle homeostasis under high-temperature conditions

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