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Oocyte maturation in mammals is characterized by a dramatic reorganization of the endoplasmic reticulum (ER). In mice, the ER forms accumulations in the germinal vesicle (GV) stage and distinctive cortical clusters in metaphase II (MII) of the oocyte. Multiple evidence suggests that this ER distribution is important in preparing the oocyte for Ca2+ oscillations, which trigger oocyte activation at fertilization. In this study, we investigated the time course and illustrated the possible functional role of ER distribution during maturation of porcine oocytes by immunostaining with protein disulfide isomerase (PDI). PDI forms clusters in the cytoplasm of oocytes. After immunostaining, PDI clusters were identified throughout the cytoplasm from the GV to metaphase I (MI) stage; however, at the MII stage, the PDI formed large clusters (1–2 µm) in the animal pole around the first polar body. PDI distribution was prevented by bacitracin, a PDI inhibitor. Our experiments indicated that, during porcine oocyte maturation, PDI undergoes a dramatic reorganization. This characteristic distribution is different from that in the mouse oocyte. Moreover, our study suggested that formation of PDI clusters in the animal pole is a specific characteristic of matured porcine oocytes. 相似文献
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Kou HIRAGA Yumi HOSHINO Kentaro TANEMURA Eimei SATO 《The Journal of reproduction and development》2013,59(4):405-408
Localization patterns of lipid droplets in the cytoplasm of porcine oocytes were
evaluated as a novel marker for in vitro maturation (IVM) of oocytes with
high developmental competence. Porcine oocytes were cultured in TCM-199, which is a
complete synthetic medium, for 44 h at 38.5 C. Localization patterns were divided into 2
classes: lipid droplets localized uniformly in the whole cytoplasm (class I) and those
that were centrally located (class II). After IVM in TCM-199, 60% of matured oocytes
exhibited the class II pattern. To investigate the relation between the distribution of
lipid droplets and the developmental rate of the oocyte, the developmental rates of class
I and class II oocytes were compared after in vitro fertilization (IVF).
Class II oocytes showed a significantly higher rate of blastocyst development than class I
oocytes. These results suggest that porcine oocytes with high developmental competence can
be selected based on the localization patterns of lipid droplets. 相似文献
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Satoshi HARA Takashi TAKANO Mio OGATA Reina YAMAKAMI Yusuke SATO Tomohiro KONO Yayoi OBATA 《The Journal of reproduction and development》2014,60(3):250-255
Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be
expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in
germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the
expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing
the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase
3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2
proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on
the 2A expression system can serve as a useful tool for examining gene function during oogenesis. 相似文献
28.
Crystalline lithocholic (3alpha-hydroxycholanic) acid was isolated from a pooled sample of feces from healthy men for the first time. This acid, which occurs in small amounts in human bile, was obtained by alcohol extraction, followed by solvent partition and chromatography. Under these conditions most of the acid was recovered in the form of its methyl ester. 相似文献
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Peng WANG Kousaku SOUMA Yuki KOBAYASHI Kei IWABUCHI Chihiro SATO Takayoshi MASUKO 《Animal Science Journal》2010,81(4):487-493
This study examined the differences between non‐inoculated (control) corn and Northern Leaf Blight (NLB)‐damaged corn (inoculated corn); dry matter (DM) yield, silage fermentation quality, nutritive value and feed intake by sheep were compared. Leaf, stem and grain dry weights and gross yield of inoculated corn were significantly (P < 0.05) decreased compared with control corn. The contents of water‐soluble carbohydrate and nitrogen‐free extract (NFE) were decreased in inoculated corn compared with control corn. Silage made from both inoculated and control corn showed good fermentation quality. The digestibility of DM, organic matter, ether extract, NFE, and energy of silage made from inoculated corn were significantly (P < 0.05) lower, and contents of total digestible nutrients (TDN) and digestible energy (DE) were also significantly (P < 0.05) lower compared with silage made from the control corn. DM intake showed no significant discrepancy between the two types of silage. TDN and DE intakes from inoculated silage were significantly (P < 0.05) lower compared with control silage. From the above results it was shown that NLB caused a decrease in DM yield and NFE content in corn and a decrease in the nutritive value and feed intake of silage. 相似文献
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Kikumi OGIHARA Ken ONDA Reiichiro SATO Yuko NAYA Hideharu OCHIAI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(1):85-88
L-type amino acid transporter
1 (LAT1), the first isotype of amino acid transport system L, transports aromatic and
branched amino acids pivotal for fundamental cellular activities such cellular growth and
proliferation. LAT1 expression was high only in the brain in contrast to its limited
distribution and low level of expression in normal tissues. We found potent LAT1
expression in canine caput epididymis by quantitative RT-PCR and Western blotting
analysis. Immnuno-histochemical examination revealed observable LAT1 in microvillous
epithelial cells. 相似文献