MSK1 regulates RANKL-induced NFATc1 expression through CREB and c-Fos

Jeongim Ha, Jung Hye Hwang, Seul Gi Kwon, Da Hye Park, Tae Wan Kim, Deok Gyeong Kang, Kyung Hee Kang, Il-Suk Kim, Chul Wook Kim 충북대학교 동물의학연구소 2015 Journal of Biomedical and Translational Research Vol.16 No.2

Osteoclasts originated from hematopoietic stem cells are multi-nucleated cells that can resorb the bone matrix. Receptor activator of nuclear factor kappa-B (RANK)/RANK ligand (RANKL) signaling pathway is crucial for the differentiation and activation of osteoclasts. In this study, we investigated for the first time whether or not RANKL induced mitogen- and stress-activated kinase 1 (MSK1) phosphorylation at Ser 376. Activation of MSK1 was detected as soon as 5 min after RANKL stimulation and sparsely detected at 30 min after stimulation. RANKL-induced MSK1 phosphorylation occurred in a dose-dependent manner. MSK1 is known as a downstream signaling molecule of cAMP-dependent protein kinase (PKA). Treatment with the PKA inhibitor H89 significantly suppressed c-Fos and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) induction upon RANKL stimulation. In addition, cAMP response element-binding protein (CREB) phosphorylation was extremely inhibited by H89 treatment. Mitogen-activated protein kinases (MAPKs) have been investigated for induction of MSK1 phosphorylation. Specific signaling pathway inhibitors for p38 and extracellular signal-regulated kinases (ERKs) significantly blocked RANKL-induced MSK1 activation. Finally, as a downstream effector of the p38-MSK1 pathway, c-Fos transcriptional activity was determined. RANKL-mediated elevation of c-Fos transcriptional activity was significantly suppressed by p38 inhibitor. Moreover, a dominant negative form of CREB suppressed activation of NFATc1. In conclusion, RANKL-stimulated MSK1 phosphorylation could play a role in induction of NFATc1 through CREB and c-Fos activation as a downstream molecule of p38, ERK MAPKs, and PKA. Our results support basic information for the development of osteoclast specific inhibitors.

MSK1 regulates RANKL-induced NFATc1 expression through CREB and c-Fos

Jeongim Ha,Jung Hye Hwang,Seul Gi Kwon,Da Hye Park,Tae Wan Kim,Deok Gyeong Kang,Kyung Hee Kang,Chul Wook Kim,Il-Suk Kim 충북대학교 동물의학연구소 2015 Journal of Biomedical and Translational Research Vol.16 No.2

Osteoclasts originated from hematopoietic stem cells are multi-nucleated cells that can resorb the bone matrix. Receptor activator of nuclear factor kappa-B (RANK)/RANK ligand (RANKL) signaling pathway is crucial for the differentiation and activation of osteoclasts. In this study, we investigated for the first time whether or not RANKL induced mitogen- and stress-activated kinase 1 (MSK1) phosphorylation at Ser 376. Activation of MSK1 was detected as soon as 5 min after RANKL stimulation and sparsely detected at 30 min after stimulation. RANKL-induced MSK1 phosphorylation occurred in a dose-dependent manner. MSK1 is known as a downstream signaling molecule of cAMP-dependent protein kinase (PKA). Treatment with the PKA inhibitor H89 significantly suppressed c-Fos and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) induction upon RANKL stimulation. In addition, cAMP response element-binding protein (CREB) phosphorylation was extremely inhibited by H89 treatment. Mitogen-activated protein kinases (MAPKs) have been investigated for induction of MSK1 phosphorylation. Specific signaling pathway inhibitors for p38 and extracellular signal-regulated kinases (ERKs) significantly blocked RANKL-induced MSK1 activation. Finally, as a downstream effector of the p38-MSK1 pathway, c-Fos transcriptional activity was determined. RANKL-mediated elevation of c-Fos transcriptional activity was significantly suppressed by p38 inhibitor. Moreover, a dominant negative form of CREB suppressed activation of NFATc1. In conclusion, RANKL-stimulated MSK1 phosphorylation could play a role in induction of NFATc1 through CREB and c-Fos activation as a downstream molecule of p38, ERK MAPKs, and PKA. Our results support basic information for the development of osteoclast specific inhibitors.

Complement component 9 (C9) 유전자의 단일염기다형성과 버크셔 돼지 육질 형질과의 연관성 분석

하정임(Jeongim Ha),황정혜(Jung Hye Hwang),유고은(Go Eun Yu),박다혜(Da Hye Park),강덕경(Deok Gyeong Kang),김태완(Tae Wan Kim),박화춘(Hwa Chun Park),안상미(Sang Mi An),김철욱(Chul Wook Kim) 한국식품과학회 2018 한국식품과학회지 Vol.50 No.5

본 연구는 Berkshire 간 조직을 이용하여 RNA-sequencing 분석을 통해 돼지 육질 연관 단일염기다형성을 발굴하기 위해 수행되었다. 그 결과, C9 유전자의 cDNA 942번 G 서열이 T 서열로 변환되어 라이신(lysin)이 아스파라진(asparagin)으로 변하는 nonsynonymous SNP를 확인하였다. Berkshire 돼지 405두에서 C9 단일염기다형성의 유전자형을 분석한 결과 major allele는 G, minor allele은 T였다. Berkshire 돼지 405두의 육질 형질을 분석하여 C9 단일염기다형성의 유전자형과 육질 형질과의 연관성 분석한 결과 우성 모델의 경우 육색의 명도, 콜라겐, 수분, 도축 후 24시간 뒤 pH (pH<SUB>24 h</SUB>) 육질 형질에서 유의성이 확인되었고, 열성 모델의 콜라겐 함량, 공우성 모델의 육색의 명도(CIE L), 단백질, 콜라겐 함량에서 유의성을 가졌다. 성별에 따른 C9 유전자형과 육질 형질 간의 연관성을 분석한 결과 거세돈에서 도체중, 콜라겐에서 유의성이 있었으며, 암퇘지의 경우 육색의 명도, 단백질, pH<SUB>24 h</SUB> 육질 형질에서 유의성이 있었다. 육질 형질 중 pH<SUB>24 h</SUB> 형질은 육질을 결정하는 중요한 형질로 C9 유전자의 유전자형이 다른 유전자형들에 비해 pH<SUB>24 h</SUB>가 증가되고 육즙 손실이 감소되는 것으로 확인되어 C9 유전자의 TG 유전자를 가진 돼지가 더 좋은 육질을 가지는 것으로 판단된다. 본 결과를 바탕으로 C9 유전자의 단일염기다형성을 육질을 판단하는 생물마커(biomarker)로 의 활용이 기대된다. In this study, to identify single nucleotide polymorphisms (SNPs) associated with meat quality in Berkshire pigs, we performed RNA sequencing. A non-synonymous SNP (nsSNP) in the Complement component 9 (C9) gene was identified, and the association between meat quality traits and the C9 genotype was analyzed. The nsSNP in the C9 gene was located at c.942 G>T. In the dominant model, significant associations were observed between the SNP and meat quality traits such as CIE L, collagen content, moisture level, and pH<SUB>24 h</SUB>, whereas in the co-dominant model, significant associations were observed between the SNP and CIE L, collagen content, and protein content. In the recessive model, a significant association between the C9 genotype and the collagen content was observed. In addition, we identified the significant relationship between the C9 genotype and meat quality according to sex. These results indicate that the C9 SNP can be used as a genetic marker for improving pork quality.

Effects of LEP, GYS1, MYOD1, and MYF5 polymorphisms on pig economic traits

Park, Sang-Je,Ha, Jeongim,Kim, Il-Suk,Kwon, Seul Gi,Hwang, Jung Hye,Park, Da Hye,Kang, Deok Gyeong,Kim, Tae Wan,Kim, Sam Woong,Kim, Chul Wook Walter de Gruyter GmbH 2015 Annals of animal science Vol.15 No.3

<B>Abstract</B><P> In the present study, we examined the effect of single nucleotide polymorphisms (SNPs) of leptin (LEP), skeletal muscle glycogen synthase (GYS1), myogenic differentiation 1 (MYOD1), and myogenic factor 5 (MYF5) genes on economic trait association in pigs. LEP/HindIII, MYOD1/DdeI, MYF5/FokI, and GYS1/FokI genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) from 466 pigs comprised of Duroc, Landrace and Yorkshire breeds. The LEP/HindIII polymorphism differed significantly with respect to average daily gain (ADG) in Duroc pigs (P<0.05). However, the GYS1/FokI polymorphism was not significantly associated with any trait. The MYOD1/DdeI polymorphism was significantly associated with both ADG and meat percentage (MP) in Duroc pigs, and ADG, backfat thickness (BFT) and feed efficiency (FE) in Landrace pigs, whereas the MYOD1/DdeI polymorphism was not significantly associated with any trait in Yorkshire pigs. In addition, the MYF5/FokI polymorphism revealed a close relationship with ADG in Duroc pigs. In conclusion, we believe that the SNPs within LEP, MYOD1 and MYF5 in certain pig breeds play important roles as potential genetic markers for economic traits of pigs.</P>

The JNK-dependent CaMK pathway restrains the reversion of committed cells during osteoclast differentiation.

Chang, Eun-Ju,Ha, Jeongim,Huang, Hao,Kim, Hyung Joon,Woo, Jung Hoon,Lee, Youngkyun,Lee, Zang Hee,Kim, Ju Han,Kim, Hong-Hee Cambridge University Press 2008 Journal of cell science Vol.121 No.15

<P>Osteoclastogenesis involves the commitment of macrophage-lineage precursors to tartrate-resistant acid phosphatase-positive (TRAP+) mononuclear pre-osteoclasts (pOCs) and subsequent fusion of pOCs to form multinuclear mature osteoclasts. Despite many studies on osteoclast differentiation, little is known about the signaling mechanisms that specifically mediate the osteoclastic commitment. In this study, we found that inhibition of JNK at the pOC stage provoked reversion of TRAP(+) cells to TRAP(-) cells. The conversion to TRAP(-) cells occurred with concomitant return to the state with higher expression of macrophage antigens, and greater activity of phagocytosis and dendritic-differentiation potential. JNK inhibition at the pOC stage reduced NFATc1 and CaMK levels, and addition of active NFATc1 partially rescued the effect of JNK inhibition. In addition, the level of NFATc1 was decreased by knockdown of CaMK by RNAi and by catalytic inhibition of CaMK, which both caused the reversion of pOCs to macrophages. These data suggest that JNK activity is specifically required for maintaining the committed status during osteoclastogenesis and that the CaMK-NFATc1 pathway is the key element in that specific role of JNK.</P>

Single-Nucleotide Polymorphisms in Pig EPHX1 Gene are Associated with Pork Quality Traits

Cho, Hwak Rae,Ha, Jeongim,Kwon, Seul Gi,Hwang, Jung Hye,Park, Da Hye,Kim, Tae Wan,Lee, Hak-Kyo,Song, Ki-Duk,Kim, Sam Woong,Kim, Chul Wook Taylor Francis 2015 Animal biotechnology Vol.26 No.3

Urinary phthalate metabolites over the first 15months of life and risk assessment – CHECK cohort study

Kim, Sunmi,Lee, Jangwoo,Park, Jeongim,Kim, Hai-Joong,Cho, Geum Joon,Kim, Gun-Ha,Eun, So-Hee,Lee, Jeong Jae,Choi, Gyuyeon,Suh, Eunsook,Choi, Sooran,Kim, Sungjoo,Kim, Sung Koo,Kim, Young Don,Kim, Su You Elsevier 2017 The Science of the total environment Vol.607 No.-

<P><B>Abstract</B></P> <P>Phthalates are important group of endocrine disruptors. Infants and young children are susceptible to phthalate exposure. However, information on the phthalate exposure during the early stages of life is very limited. This study was conducted to understand the temporal trend of exposure to major phthalates among infants of Korea during the first 15months after birth, and to estimate associated risks. A total of 286 urine samples were collected from 171 children at 3, 9, 12, or 15months of age, with 77 children sampled for two or more times. Four phthalates, i.e., di(2-ethylhexyl) phthalate (DEHP), di-isobutyl phthalate (DiBP), di-n-butyl phthalate (DnBP), and diethyl phthalate (DEP) were chosen, and their major metabolites were analyzed in the urine. The DEHP metabolites were detected in 100% of the urine samples at relatively higher levels compared to those reported in other countries. The levels of mono-ethyl phthalate (MEP) were generally lower. Urinary concentrations of most phthalate metabolites, especially DEHP metabolites, increased as children grew older. Intra-class correlation coefficients (ICCs) calculated for DEHP metabolites over time were high (0.7–0.8), suggesting persistence of consistent exposure sources during this sensitive period of life. Hazard quotient (HQ) and hazard index (HI) were calculated from daily intake estimates divided by recommended toxicity thresholds. Among the study population, 4, 16, and 26% of the children showed HI >1 at 9, 12, and 15months of age, respectively. DEHP exposure explained most of the risk estimates. Considering vulnerability of young children to endocrine disruption, efforts to identify sources of exposure and to develop appropriate mitigation options are warranted.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Multiple urines were collected at 3, 9, 12, or 15months of age from 171 children. </LI> <LI> High intra-individual correlation and age-dependent increase were observed. </LI> <LI> Up to 26% of children showed hazard index >1 by exposure to target phthalates. </LI> <LI> Estimated risks are mostly due to the exposure to DEHP and DnBP. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Identification of a Bromodomain-containing Protein 2 (BRD2) Gene Polymorphic Variant and Its Effects on Pork Quality Traits in Berkshire Pigs

Dong Ju Lee,Jung Hye Hwang,Jeongim Ha,Go Eun Yu,Seulgi Kwon,Da Hye Park,Deok Gyeong Kang,Tae Wan Kim,Hwa Chun Park,Sang Mi An,Chul Wook Kim 한국축산식품학회 2018 한국축산식품학회지 Vol.38 No.4

Bromodomain-containing protein 2 (BRD2) is a nuclear serine/threonine kinase involved in transcriptional regulation. We investigated the expression and association of the BRD2 gene as a candidate gene for meat quality traits in Berkshire pigs. BRD2 mRNA was expressed at relatively high levels in muscle tissue. Statistical analysis revealed that the c.1709G>C polymorphism of the BRD2 gene was significantly associated with carcass weight, meat color (a*, redness), protein content, cooking loss, water-holding capacity, carcass temperatures 4, 12 and 24 h postmortem, and the 24 h postmortem pH in 384 Berkshire pigs. Therefore, this polymorphism in the porcine BRD2 gene may be used as a candidate genetic marker to improve meat quality traits in pigs.

Bisphenol A distribution in serum, urine, placenta, breast milk, and umbilical cord serum in a birth panel of mother–neonate pairs

Lee, Jangwoo,Choi, Kyungho,Park, Jeongim,Moon, Hyo-Bang,Choi, Gyuyeon,Lee, Jeong Jae,Suh, Eunsook,Kim, Hai-Joong,Eun, So-Hee,Kim, Gun-Ha,Cho, Geum Joon,Kim, Sung Koo,Kim, Sungjoo,Kim, Su Young,Kim, Se Elsevier 2018 Science of the Total Environment Vol.626 No.-

<P><B>Abstract</B></P> <P>Bisphenol A (BPA) exposure during the perinatal and postnatal periods increases the susceptibility to disease over the life cycle. However, information on the BPA delivered to fetuses or infants via the placenta and breastfeeding is limited. We determined the BPA exposure levels in various bodily fluids and tissues of pregnant women and described fetus and infant exposures to BPA based on associations and BPA ratios in mother–neonate paired samples. Maternal serum, urine, placenta, breast milk, cord serum, and neonatal urine samples were collected from 318 mother–neonate pairs at six university hospitals in Korea. BPA levels were detected using liquid chromatography tandem mass spectrometry. The ratios of the BPA levels in the other sample types to the levels in maternal serum were calculated. BPA was detected in 79.5–100% of the maternal and fetal samples. The median BPA concentration in the samples decreased in the order of neonatal urine (4.75ng/mL), maternal urine (2.86ng/mL), cord serum (1.71ng/mL), maternal serum (1.56ng/mL), breast milk (0.74ng/mL), and the placenta (0.53ng/g). We estimated the ratios of BPA levels in the other sample types to those in maternal serum. The median (95th percentile) cord serum-to-maternal serum ratio was 1.12 (15.2) for 160 mother–fetal pairs, in which BPA was detected in both samples. The placenta-, maternal urine-, neonatal urine-, and breast milk-to-maternal serum ratios were 0.28 (5.31), 1.79 (29.9), 1.98 (28.2), and 0.51 (10.5), respectively. In addition, the median (95th percentile) cord serum-to-placenta ratio was 4.03 (45.8), and the neonatal urine-to-cord serum ratio was 1.95 (25.6). The 95th percentile values were 14–20-fold greater than the medians. Urine contained the highest BPA concentrations, followed by serum, breast milk, and the placenta. The variations of BPA ratio show individual differences in the amounts of BPA delivered from mother to fetus.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Not enough data on tissue distribution of BPA in mother‑neonate (or fetus) pair </LI> <LI> The order of BPA concentrations in examined tissue or bio-samples are urine in mother and neonates>cord serum>maternal serum>breast milk>placenta. </LI> <LI> BPA in cord serum, significantly associated with in maternal serum and urine but not in others. </LI> <LI> The variations of BPA ratio show individual differences in the amounts of BPA delivered from mother to fetus. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

TLT-1s, Alternative Transcripts of Triggering Receptor Expressed on Myeloid Cell-like Transcript-1 (TLT-1), Inhibits the Triggering Receptor Expressed on Myeloid Cell-2 (TREM-2)-mediated Signaling Pathway during Osteoclastogenesis

Yoon, Soo-Hyun,Lee, Yong Deok,Ha, Jeongim,Lee, Youngkyun,Kim, Hong-Hee American Society for Biochemistry and Molecular Bi 2012 The Journal of biological chemistry Vol.287 No.35