The advanced paraffin-section preparation technique based on multiple cumulus-oocyte complexes rather than ovaries in ovine

Namei, Erge; Sun, Wei; Pan, Deng; Zhao, Yufen; Yang, Bo-sheng; Yu, Wei; Du, Chenguang; Li, Haijun; Yu, Boyang; Subudeng, Gerile

doi:10.1016/j.repbio.2020.100473

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…Morphological Quantitative Analysis. The process of making paraffin sections refers to the research of Namei et al [38]. The white muscle tissues after fixation in fixative (4% paraformaldehyde solution) for 24 h were dehydrated with 75%, 85%, 95%, and 100% absolute ethanol in sequence, then transparent with xylene, and immersed in wax for 2.5 h. The paraffin-impregnated tissues were embedded in a machine from Leica, Berlin, Germany, and after solidification, then a thickness of 6 μm paraffin sections were made in a microtome (Leica RM2016, Berlin, Germany).…”

Section: Methodsmentioning

confidence: 99%

Dietary Leucine Supplementation Improves Muscle Fiber Growth and Development by Activating AMPK/Sirt1 Pathway in Blunt Snout Bream (Megalobrama amblycephala)

Wang

Guo

Huang

et al. 2022

Aquaculture Nutrition

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This research is aimed at evaluating the effects of leucine supplementation on muscle fibers growth and development of blunt snout bream through a feeding trial and a primary muscle cells treatment. An 8-week trial with diets containing 1.61% leucine (LL) or 2.15% leucine (HL) was conducted in blunt snout bream ( mean initial weight = 56.56 ± 0.83 g ). Results demonstrated that the specific gain rate and the condition factor of fish in the HL group were the highest. The essential amino acids content of fish fed HL diets was significantly higher than that fed LL diets. The texture (hardness, springiness, resilience, and chewiness), the small-sized fiber ratio, fibers density, and sarcomere lengths in fish all obtained the highest in the HL group. Additionally, the proteins expression related with the activation of the AMPK pathway (p-Ampk, Ampk, p-Ampk/Ampk, and Sirt1) and the expression of genes (myogenin (myog), myogenic regulatory factor 4 (mrf4) and myoblast determination protein (myod), and protein (Pax7) related to muscle fiber formation were significantly upregulated with increasing level of dietary leucine. In vitro, the muscle cells were treated with 0, 40 and 160 mg/L leucine for 24 h. The results showed that treated with 40 mg/L leucine significantly raised the protein expressions of BCKDHA, Ampk, p-Ampk, p-Ampk/Ampk, Sirt1, and Pax7 and the gene expressions of myog, mrf4, and myogenic factor 5 (myf5) in muscle cells. In summary, leucine supplementation promoted muscle fibers growth and development, which may be related to the activation of BCKDH and AMPK.

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Section: Methodsmentioning

confidence: 99%

Dietary Leucine Supplementation Improves Muscle Fiber Growth and Development by Activating AMPK/Sirt1 Pathway in Blunt Snout Bream (Megalobrama amblycephala)

Wang

Guo

Huang

et al. 2022

Aquaculture Nutrition

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“…The localization of CX43 protein in immature COCs was detected using a novel COCs paraffin section technique developed in our laboratory ( 31 ). The findings revealed exclusive CX43 expression in CCs, with no detectable CX43 expression in oocytes ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In this study, we first identified the location of the CX43 protein in immature sheep COCs using a novel paraffin section technique developed in our laboratory ( 31 ). The results revealed that the CX43 protein was exclusively present in the cumulus compartment of sheep COCs.…”

Section: Discussionmentioning

confidence: 99%

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Cyclic AMP mediates ovine cumulus–oocyte gap junctional function via balancing connexin 43 expression and phosphorylation

Zhao,

Namei,

Yang

et al. 2023

Endocrine Connections

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Gap junction channels in cumulus-oocyte complexes (COCs) enable the transmission and communication of small molecular signals between adjacent cells, such as cyclic adenosine monophosphate (cAMP). However, the regulation of gap junction function (GJF) by cAMP and the underlying mechanisms involved are not fully clarified. This study investigated the effect of cAMP on CX43 expression and GJF in ovine COCs using immunofluorescence, quantitative real-time PCR (qRT-PCR), western blotting and GJF detection. The CX43 was only found in the cumulus cells (CCs) side of ovine COC. The intra-oocyte cAMP showed a significant increase at 30 min, while the intra-CC cAMP exhibited two peaks at 10 min and 1 h during in vitro maturation (IVM). Phosphorylated CX43 protein exhibited an immediate increase at 10 min, and CX43 protein displayed two peaks at 10 min and 1 h during IVM. The duration of pre-IVM exposure to forskolin and IBMX significantly enhanced phosphorylated and total Cx43, as well as Gja1 and Creb genes for 10 min; these effects were counteracted by Rp-cAMP. Pre-IVM with forskolin and IBMX for 1 h, the GJF and CX43/p-CX43 ratio were both elevated. The closure of gap junction channels caused by phosphorylated CX43 to prevent cAMP outflow from oocytes in early IVM of COC. Cyclic AMP up-regulated phosphorylated and total Cx43 via genomic and non-genomic pathways, but its functional regulation was dependent on the balance of the two proteins. This study provides a new insight into the regulatory mechanism between cAMP and GJF, which would improve IVM in animal and clinical research.

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The ERβ-cAMP signaling pathway regulates estradiol-induced ovine oocyte meiotic arrest

Yang,

An,

Yang

et al. 2024

Theriogenology

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The advanced paraffin-section preparation technique based on multiple cumulus-oocyte complexes rather than ovaries in ovine

Cited by 3 publications

References 16 publications

Dietary Leucine Supplementation Improves Muscle Fiber Growth and Development by Activating AMPK/Sirt1 Pathway in Blunt Snout Bream (Megalobrama amblycephala)

Dietary Leucine Supplementation Improves Muscle Fiber Growth and Development by Activating AMPK/Sirt1 Pathway in Blunt Snout Bream (Megalobrama amblycephala)

Cyclic AMP mediates ovine cumulus–oocyte gap junctional function via balancing connexin 43 expression and phosphorylation

The ERβ-cAMP signaling pathway regulates estradiol-induced ovine oocyte meiotic arrest

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