• Development and Reproduction
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• v.23 no.1
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• pp.1-9
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• 2019

The ability of oocytes to undergo normal fertilization and embryo development is acquired during oocyte maturation which is transition from the germinal vesicle stage (GV), germinal vesicle breakdown (GVBD) to metaphase of meiosis II (MII). Part of this process includes redistribution of inositol 1, 4, 5-triphosphate receptor (IP3R), a predominant $Ca^{2+}$ channel on the endoplasmic reticulum membrane. Type 1 IP3R (IP3R1) is expressed in mouse oocytes dominantly. At GV stage, IP3R1 are arranged as a network throughout the cytoplasm with minute accumulation around the nucleus. At MII stage, IP3R1 diffuses to the entire cytoplasm in a more reticular manner, and obvious clusters of IP3R1 are observed at the cortex of the egg. This structural reorganization provides acquisition of $[Ca^{2+}]_i$ oscillatory activity during fertilization. In this review, general properties of IP3R1 in somatic cells and mammalian oocyte are introduced.

• YAKHAK HOEJI
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• v.54 no.1
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• pp.8-12
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• 2010

Inositol 1,4,5-trisphosphate ($IP_3$) receptor ($IP_3R$)-mediated signaling pathway is involved in many cellular processes including fertilization, apoptosis and neuronal function. Although cardiac myocytes express the $IP_3R$, its pathophysiological role has not been clearly understood because of limited selectivity of currently available pharmacological blockers. In the present study we constructed shRNA-expressing adenovirus to knock-down the type 2 $IP_3R$ ($IP_3R2$), a major subtype in cardiac ventricular myocytes, and demonstrated that the virus successfully eliminated the expression and localization of the $IP_3R2$. These results may provide a reliable tool for probing pathophysiological roles of the $IP_3R2$ in isolated intact cardiac myocytes.

• Journal of Animal Science and Technology
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• v.51 no.4
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• pp.289-294
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• 2009

Inositol 1,4,5-triphosphate receptor type 1 (IP3R1) is a $Ca^{2+}$ release channel that responds to the second messenger IP3 and that modulates diverse cellular functions such as contraction/excitation, secretion, gene expression and cellular growth. We discovered single nucleotide polymorphisms (SNPs) within IP3R1 gene and analyzed associations between gene polymorphisms and carcass traits in Korean cattle (Hanwoo) in order to develop novel DNA markers at genomic level. Three SNPs were detected at the position of g.1428617A>G, g.1418843C>T and g.1414377C>T with 24 unrelated Hanwoo samples by direct sequencing of the PCR products. We found that genotype of g.1414377C>T SNP was associated with live weight (P<0.05) and carcass weight (P<0.01) using the general linear model of SAS package. These results suggest that polymorphism of IP3R1 gene was associated with weight-related traits in Hanwoo.

• Journal of the Korean Institute of Landscape Architecture
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• v.29 no.4
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• pp.82-90
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• 2001

This study was carried out to reveal optimum conditions for in vitro propagation of 3 Korean native azalea species, Rhododendron mucronulatum, R. yedonese var. poukhanense, and R. shlippenbachii, which are useful for landscape proposes. Seeds and meristems from three azalea species were cultured on 1/2MS, Hyponex, and Anderson media containing growth of regulators benzyladenine(BA) and 2-isopentenyadenosine(2ip). The results were as follows. 1. In the culture of R. schlippenbachii and R. mucronulatum seeds, in vitro seedlings germinated and grew well on he 1/2MS and Anderson media, while R. yedoense var. poukhanense on Hyponex media containing 6.0mg/$\ell$ 2ip. 2. When the meristems of R. mucronulatum were cultured on Andeson media containing 9.0mg/$\ell$ 2ip, the survival rate of meristems was 23.0% in 6 weeks after culture, and the survival rate of R. schlippenbachii was 46.0% o nthe same media containing 12.0mg/$\ell$ 2ip. The survial rate of R. yedoense var. poukhanense was 92.0% onHyponex media containing 0.5mg/$\ell$ BA and 9.0mg/$\ell$ 2ip. When the meristems of R. mucronulatum and R. yedoense var. poukhanense were cultured on Hyponex media containing 12.0mg/$\ell$ 2ip, they showed the most excellent growth. R. schlippenbachii grew well on Anderson media containing 9.0mg/$\ell$ 2ip. When in vitro shoots of R. yedoense var. poukhanense were subcultured to solid medium, they grew well in shoot growth on Hyponex media containing 6.0mg/$\ell$ 2ip.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.20
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• pp.8595-8601
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• 2014

Background: miR-200a expression is frequently altered in numerous cancers. The aim of the present study was to determine the role of microRNA-200a in advanced ovarian carcinomas. Materials and Methods: We measured miR-200a expression in 72 matched normal ovarian tissues and advanced ovarian carcinomas, and also two ovarian carcinoma cell lines (SKOV3 and SKOV3.ip1 - the latter being more invasive and metastatic than the parental SKOV3) by stem-loop real-time RT-PCR based on TaqMan microRNA assay using U6 as a reference. Levels of miR-200a expression were compared by disease stage, tumor grade, histology, and lymph node involvement. To evaluate the role of microRNA-200a, cell proliferation and invasion of SKOV-3 and SKOV-3.ip1 were analyzed with miR-200a inhibitor/mimic transfected cells. Results: Of 72 paired samples, 65 cancer tissues overexpressed microRNA-200a greater than two fold in comparison with matched normal epithelium. Specifically, patients with lymph node metastasis showed significant elevation. The level correlated with clinicopathological features, including high tumor grade, late disease stage, most notably with lymph node metastasis, but not with tumor histology. In addition, SKOV-3.ip1 cells also overexpressed miR-200a compared with SKOV-3, and miR-200a inhibitor transfected SKOV-3.ip1 cells showed significant reduction in cellular proliferation and invasion, while a miR-200a mimic stimulated the opposite behavior. Conclusions: We provide definitive evidence that miR-200a is up-regulated in a significant proportion of advanced ovarian carcinomas, and that elevated miR-200a expression facilitates tumor progression. Our findings support the notion that miR-200a is an onco-microRNA for ovarian cancer, and elevation is a useful potential diagnostic indicator. This study also provides a solid basis for further functional analysis of miR-200a in advanced ovarian cancer.

• Genomics & Informatics
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• v.19 no.2
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• pp.17.1-17.13
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• 2021

Breast cancer is one of the leading causes of cancer in women all over the world and accounts for ~25% of newly observed cancers in women. Epigenetic modifications influence differential expression of genes through non-coding RNA and play a crucial role in cancer regulation. In the present study, epigenetic regulation of gene expression by in-silico analysis of histone modifications using chromatin immunoprecipitation sequencing (ChIP-Seq) has been carried out. Histone modification data of H3K4me3 from one normal-like and four breast cancer cell lines were used to predict miRNA expression at the promoter level. Predicted miRNA promoters (based on ChIP-Seq) were used as a probe to identify gene targets. Five triple-negative breast cancer (TNBC)-specific miRNAs (miR153-1, miR4767, miR4487, miR6720, and miR-LET7I) were identified and corresponding 13 gene targets were predicted. Eight miRNA promoter peaks were predicted to be differentially expressed in at least three breast cancer cell lines (miR4512, miR6791, miR330, miR3180-3, miR6080, miR5787, miR6733, and miR3613). A total of 44 gene targets were identified based on the 3'-untranslated regions of downregulated mRNA genes that contain putative binding targets to these eight miRNAs. These include 17 and 15 genes in luminal-A type and TNBC respectively, that have been reported to be associated with breast cancer regulation. Of the remaining 12 genes, seven (A4GALT, C2ORF74, HRCT1, ZC4H2, ZNF512, ZNF655, and ZNF608) show similar relative expression profiles in large patient samples and other breast cancer cell lines thereby giving insight into predicted role of H3K4me3 mediated gene regulation via the miRNA-mRNA axis.

• The Korean Journal of Food And Nutrition
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• v.9 no.2
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• pp.123-136
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• 1996

IPSKCDNA gene(1.8 kbp) encoding rat brain IP3K enzyme contained Not I restric site in open reading frame. The Not I sequence, GCGGCCGC, was converted to GCAGCCGC by site-directed mutagenesis. The mutated IP3KcDNA was digested with EcoR I and ligated with EcoR I-restricted psp72·Not2 vector. The resulting psp72 · Not2-IP3KCDNA was digested with the Not I restriction enzyme and then subcloned into the Not I -digested PZIP · NeoSV(X) mammalian expression vector. The PZIP · NeoSV(X) -IPSKCDNA was transfected into CCL39 hamster lung fibroblast cells. The efficiency of the expressed IPSKCDNA gene was significantly higher than expected generally, not only a mean 5-fold increase in the amount of enzyme, but also 16-fold increase in enzyme activity from tractsfected CCL39 cells by the method of Western blot using anti-lP3K antibodies. Both distribution of IPSK in various rat tissues and biochemical properties were discussed.

• Molecules and Cells
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• v.22 no.3
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• pp.328-335
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• 2006

Imperatoxin A ($IpTx_a$), a 3.7 kDa peptide from the African scorpion Pandinus imperator, is an agonist of the skeletal muscle ryanodine receptor (RyR1). In order to study the structure of the toxin and its effect on RyR1, $IpTx_a$ cDNA was PCR-amplified using 3 pairs of primers, and the toxin was expressed in E. coli. The toxin was further purified by chromatography, and various point mutants in which basic amino acids were substituted by alanine were prepared by site-directed mutagenesis. Studies of single channel properties by the planar lipid bilayer method showed that the recombinant $IpTx_a$ was identical to the synthetic $IpTx_a$ with respect to high-performance liquid chromatography mobility, amino acid composition and specific effects on RyR1. Mutations of certain basic amino acids ($Lys^{19}$, $Arg^{23}$, and $Arg^{33}$) dramatically reduced the capacity of the peptide to activate RyRs. A subconductance state predominated when $Lys^8$ was substituted with alanine. These results suggest that some basic amino acid residues in $IpTx_a$ are important for activation of RyR1, and that $Lys^8$ plays an important role in regulating the gating mode of RyR1.

• Biomedical Science Letters
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• v.13 no.4
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• pp.333-339
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• 2007

Recently, much attention has been paid to human retinoblastoma since it provide a good model system for studying mechanisms underlying cell growth, differentiation, proliferation, and apoptosis, and for developing cancer therapy. However, until now it is unclear whether purinergic receptors are involved in the calcium mobilization in the retinoblastoma cells. In this regard, we measured possible purinergic signaling in WERI-Rb-1 cells using $Ca^{2+}$ imaging technique and RT-PCR method. ATP-induced $[Ca^{2+}]_i$ transients was maintained to about $90.7{\pm}1.0%$ of the control (n=48) even in the absence of extracellular calcium. The ATP-induced intracellular calcium response was only attained to $10.4{\pm}1.8%$ (n=55) of peak amplitude of the control after preincubation of 1 ${\mu}MU-73122$, a PLC inhibitor, but it was not affected by 1 ${\mu}MU-73343$, a inactive form of U-73122. And also ATP-induced $[Ca^{2+}]_i$ rise was almost attenuated by 20 ${\mu}M$ 2-APB, a putative $IP_3$ receptor inhibitor. Two subtypes of $IP_3$ receptor $(IP_{3-1}R,\;IP_{3-2}R)$ were identified by a RT-PCR method. These findings suggest that purinergic stimuli can cause calcium mobilization via $PLC-IP_3$ pathway after the activation of P2Y receptors in the retinoblastoma cells, which may play important roles in cell proliferation, differentiation, growth, and cell death.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.215-225
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• 2003

This study was carried out to investigate the change of blood compositions by age in Hanwoo, and a total of 866 of Hanwoo, which consisted with 638 of steer and 228 of bulls, were used to measure serum concentrations. A multiple regression equation was estimated with collection age and blood composition as independent and dependent variables, respectively. Complicated regression equations for blood compositions in steer and bulls were IGF-I(cubic), calcium (linear), and IP(linear). Linear and cubic equations were fitted to testosterone in steer and creatinine in bulls, respectively. A cubic equation in steer and linear equation in bulls were fitted to HDLC. Equations of quadratic in steer and cubic in bulls were fitted to concentration of triglyceride, globulin, and A/G ratio. BUN was fitted by equations of cubic in steer and quadratic in bulls. TP and albumin were fitted by equations of quadratic in steer and linear in bulls. A cubic regression equation did not explain the change of cortisol by age in steer and bulls. A cubic regression equation did explain the change of glucose by age in steer, but not in bulls. Higher R-square values (R-SQUARE>0.1) were estimated to IGF-1, albumin, creatinine, Inorganic phosphorous(IP) and HDLC in steer, and testosterone, IGF-I, TP, albumin, glucose, creatinine, IP, and HDLC in bulls for the fitted regression equations of blood compositions. Therefore, IGF-I, albumin, creatinine, IP, and HDLC were regarded as comparatively large variation by age in steer and bulls.