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Jun-Koo Yi,
Hei-Jung Kim,
Dong-Hoon Yu,
Seo-Jin Park, Mi-Jung Shin,
Hyung-Soo Yuh,
Ki-Beom Bae,
Young-Rae Ji,
Na-Ri Kim,
Si-Jun Park,
Jae-Young Kim,
Hyun-Shik Lee,
Sang-Gyu Lee,
Du Hak Yoon,
Byung-Hwa Hyun,
Wan-Uk Kim,
Zae-Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: Calcineurin-binding protein 1 (CABIN-1) regulates calcineurin phosphatase activity as well as the activation, apoptosis, and inflammatory responses of fibroblast-like synoviocytes (FLS), which actively participate in the chronic inflammatory responses in rheumatoid arthritis (RA). However, the mechanism of action of CABIN-1 in FLS apoptosis is not clear. This study was undertaken to define the regulatory role of CABIN-1 in FLS from mice with collagen-induced arthritis (CIA).
Transgenic mice overexpressing human CABIN-1 in joint tissue under the control of a type II collagen promoter were generated. Expression of human CABIN-1 (hCABIN-1) in joints and FLS was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of cytokines, matrix metalloproteinases (MMPs), and apoptosis-related genes in FLS was determined by enzyme-linked immunosorbent assay, gelatin zymography, and RT-PCR, respectively. Joints were stained with hematoxylin and eosin and with tartrate-resistant acid phosphatase for histologic analysis.
Human CABIN-1-transgenic mice with CIA had less severe arthritis than wild-type mice with CIA, as assessed according to hind paw thickness and histologic features. The milder arthritis was accompanied by significantly enhanced apoptosis in transgenic mice, evidenced by a significantly greater number of TUNEL-positive cells in synovial tissue. Expression of inflammatory cytokines and MMPs in the transgenic mice with CIA was reduced, and they exhibited decreased Akt activation and increased expression of p53, caspase 3, caspase 9, and Bax.
Our findings demonstrate that hCABIN-1 plays a critical role in promoting apoptosis of FLS and in attenuating inflammation and cartilage and bone destruction in RA. These results help elucidate the pathogenic mechanisms of RA and suggest that CABIN-1 is a potential target for treatment of this disease.
Arthritis & Rheumatism 01/2012; 64(7):2191-200. · 7.87 Impact Factor
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Hei Jung Kim,
Young Rae Ji,
Myoung Ok Kim,
Dong Hoon Yu, Mi Jung Shin,
Hyung Soo Yuh,
Ki Beom Bae,
Seo jin Park,
Jun Koo Yi,
Na Ri Kim,
Si Jun Park,
Du Hak Yoon,
Won-Ha Lee,
Sanggyu Lee,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: The T-cell receptor (TCR) engages with an antigen and initiates a signaling cascade that leads to the activation of transcription factors. Roquin, a protein encoded by the RC3H1 gene and characterized as an immune regulator, was recently identified as a novel RING-type ubiquitin ligase family member, but the mechanisms by which Roquin regulates T-cell responses are unclear. We used the EL-4 murine lymphoma cell line to elucidate the role of Roquin in vitro. Roquin-overexpressing EL-4 cells became hyper-responsive after anti-CD3/CD28 stimulation in vitro and were a major source of the cytokines IL-2 and TNF-α. Upon activation, these cells showed particularly enhanced production of IL-2 and TNF-α. To clarify the important role played by Roquin in T-cell responses ex vivo, we generated T-cell-specific Roquin transgenic (Tg) mice. Roquin-Tg CD4(+) T-cells showed enhanced production of IL-2 and TNF-α in response to TCR stimulation with anti-CD28 co-stimulation. Further studies are necessary to investigate the role of Roquin in the regulation of primary T-cell activation, survival, and differentiation.
Biochemical and Biophysical Research Communications 11/2011; 417(1):280-6. · 2.48 Impact Factor
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Ki Beom Bae,
Myoung Ok Kim,
Dong Hoon Yu, Mi Jung Shin,
Hei Jung Kim,
Hyung Soo Yuh,
Young Rae Ji,
Jae-Young Kim,
Jin Man Kim,
Byung Hwa Hyun,
Hwi Cheul Lee,
Won Kyong Chang,
Soo Bong Park,
Do Hyung Kim,
Hyun-Shik Lee,
Yeon-Sik Choo,
Sanggyu Lee,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: The transcription factor Juxtaposed with another zinc finger gene 1 (JAZF1) is a zinc finger protein that binds to the nuclear orphan receptor TR4. Recent evidence indicates that TR4 receptor functions as both a positive and negative regulator of transcription, but the role of JAZF1 in transcriptional mechanisms has not been elucidated. Recently, the incidence rate of congenital heart malformations was reported to be significantly elevated in patients who had neurofibromatosis 1 (NF1) with chromosomal microdeletion syndrome. Furthermore, Joined to JAZF1 (SUZ12) is expressed at high levels in the hearts of adult patients with NF1 microdeletion syndrome. Therefore, we hypothesized that ectopic expression of JAZF1 may lead to cardiac malformations that deleteriously affect the survival of neonates and adults. We sought to elucidate the role of JAZF1 in cardiac development using a Jazf1-overexpressing (Jazf1-Tg) mouse model. In Jazf1-Tg mice, Jazf1 mRNA expression was significantly elevated in the heart. Jazf1-Tg mice also showed cardiac defects, such as high blood pressure, electrocardiogram abnormalities, apoptosis of cardiomyocytes, ventricular non-compaction, and mitochondrial defects. In addition, we found that the expression levels of pro-apoptotic genes were elevated in the hearts of Jazf1-Tg mice. These findings suggest that Jazf1 overexpression may induce heart failure symptoms through the upregulation of pro-apoptotic genes in cardiomyocytes.
Transgenic Research 01/2011; 20(5):1019-31. · 2.75 Impact Factor
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Young Rae Ji,
Myoung Ok Kim,
Sung Hyun Kim,
Dong Hun Yu, Mi Jung Shin,
Hei Jung Kim,
Hyung Soo Yuh,
Ki Beom Bae,
Jae Young Kim,
Hum Dai Park,
Sang Gyu Lee,
Byung Hwa Hyun,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: Wnt/Wg genes play a critical role in the development of various organisms. For example, the Wnt/beta-catenin signal promotes heart formation and cardiomyocyte differentiation in mice. Previous studies have shown that RGS19 (regulator of G protein signaling 19), which has Galpha subunits with GTPase activity, inhibits the Wnt/beta-catenin signal through inactivation of Galpha(o). In the present study, the effects of RGS19 on mouse cardiac development were observed. In P19 teratocarcinoma cells with RGS19 overexpression, RGS19 inhibited cardiomyocyte differentiation by blocking the Wnt signal. Additionally, several genes targeted by Wnt were down-regulated. For the in vivo study, we generated RGS19-overexpressing transgenic (RGS19 TG) mice. In these transgenic mice, septal defects and thin-walled ventricles were observed during the embryonic phase of development, and the expression of cardiogenesis-related genes, BMP4 and Mef2C, was reduced significantly. RGS19 TG mice showed increased expression levels of brain natriuretic peptide and beta-MHC, which are markers of heart failure, increase of cell proliferation, and electrocardiogram analysis shows abnormal ventricle repolarization. These data provide in vitro and in vivo evidence that RGS19 influenced cardiac development and had negative effects on heart function.
Journal of Biological Chemistry 09/2010; 285(37):28627-34. · 4.77 Impact Factor
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Young Rae Ji,
Myoung Ok Kim,
Sung Hyun Kim,
Dong Hun Yu, Mi Jung Shin,
Hei Jung Kim,
Hyung Soo Yuh,
Ki Beom Bae,
Jae Young Kim,
Hum Dai Park,
Sang Gyu Lee,
Byung Hwa Hyun,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: Wnt/Wg genes play a critical role in the development of various organisms. For example, the Wnt/β-catenin signal promotes
heart formation and cardiomyocyte differentiation in mice. Previous studies have shown that RGS19 (regulator of G protein signaling 19), which has Gα subunits with GTPase activity, inhibits the Wnt/β-catenin signal through inactivation of Gαo. In the present study, the effects of RGS19 on mouse cardiac development were observed. In P19 teratocarcinoma cells with
RGS19 overexpression, RGS19 inhibited cardiomyocyte differentiation by blocking the Wnt signal. Additionally, several genes
targeted by Wnt were down-regulated. For the in vivo study, we generated RGS19-overexpressing transgenic (RGS19 TG) mice. In these transgenic mice, septal defects and thin-walled
ventricles were observed during the embryonic phase of development, and the expression of cardiogenesis-related genes, BMP4
and Mef2C, was reduced significantly. RGS19 TG mice showed increased expression levels of brain natriuretic peptide and β-MHC,
which are markers of heart failure, increase of cell proliferation, and electrocardiogram analysis shows abnormal ventricle
repolarization. These data provide in vitro and in vivo evidence that RGS19 influenced cardiac development and had negative effects on heart function.
Journal of Biological Chemistry 09/2010; 285(37):28627-28634. · 4.77 Impact Factor
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Mi Jung Shin,
Jeong-Han Lee,
Dong Hoon Yu,
Hye Jung Kim,
Ki Beom Bae,
Hyung Soo Yuh,
Myoung Ok Kim,
Byung-Hwa Hyun,
Sanggyu Lee,
Raekil Park,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: The circling (cir/cir) mouse is a murine model for human nonsyndromic deafness DFNB6. Transmembrane inner ear (tmie) is the causative gene and its mutation through deletion of a 40-kilobase genomic region including tmie leads to deafness. The function of Tmie is unknown. To better understand the function of Tmie, we focused on the spatiotemporal expression of tmie in the rat cochlea by using a Tmie-specific antibody. Results showed that tmie expression was prominent in early postnatal rat cochleas in the stereocilia bundles of hair cells. The Tmie signal spread from the stereocilia to the hair cell body region and on to organ of Corti cells. No Tmie signal was observed in cell nuclei; Tmie was localized to the cytoplasm. Because Tmie is predicted to have 1 or 2 transmembrane domains, we postulate that it is localized to membrane-based organelles or the plasma membrane. Our results imply that Tmie exists in the cytoplasm and may have a key role in the maturation and structure of stereocilia bundles in developing hair cells. After hair cell maturation, Tmie is thought to be involved in the maintenance of organ of Corti cells.
Comparative medicine 08/2010; 60(4):288-94. · 1.05 Impact Factor
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Mi Jung Shin,
Jeong-Han Lee,
Dong Hoon Yu,
Bong Soo Kim,
Hei Jung Kim,
Sung Hyun Kim,
Myoung Ok Kim,
Channy Park,
Byung-Hwa Hyun,
Sanggyu Lee,
Hong-Seob So,
Raekil Park,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: The circling (cir/cir) mouse is one of the murine models for human non-syndromic deafness DFNB6. The mice have abnormal circling behavior, suggesting a balanced disorder and profound deafness. The causative gene was transmembrane inner ear (tmie) gene of which the mutation is a 40-kb genomic deletion including tmie gene itself. In this study, tmie-overexpression trasngenic mice were established. Individuals with germline transmission have been mated with circling homozygous mutant mice (cir/cir) in order to produce the transgenic mutant mice (cir/cir-tg) as a gene therapy. After the genotyping, phenotypic analyses were performed so that the insertion of the new gene might compensate for the diseases such as hearing loss, circling behavior, or swimming inability. Some individuals exhibited complete recovery in their behavior and hearing but the others did not show any amelioration in behavior or hearing. Individual mice had very different levels of tmie transgene expression in the cochlea. These results clearly indicate that tmie protein plays an important role when the appropriate expression level of tmie was expressed in the inner ear. The protein levels were variable in each individual and these are thought to induce the differences in disease amelioration levels.
Biochemical and Biophysical Research Communications 07/2008; 374(1):17-21. · 2.48 Impact Factor
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Dong Hoon Yu,
Myoung Ok Kim,
Sung Hyun Kim, Mi Jung Shin,
Bong Soo Kim,
Hei Jung Kim,
Sang Ryeul Lee,
Sang Gyu Lee,
Seung-Ah Yoo,
Wan Uk Kim,
Byung Hwa Hyun,
Young Sik Park,
Tae Yoon Kim,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: Rheumatoid arthritis is a chronic inflammatory disease. The generation of reactive oxygen species (ROS) within an inflamed joint has been suggested as playing a significant pathogenic role. Extracellular superoxide dismutase (EC-SOD) is a major scavenger enzyme of ROS, which has received growing attention for its therapeutic potential. To investigate the therapeutic effect of EC-SOD in mice with collagen-induced arthritis (CIA), we used mouse embryonic fibroblast (MEF) of transgenic mice that overexpresses EC-SOD on the skin by using hK14 promoter. DBA/1 mice that had been treated with bovine type II collagen were administrated subcutaneous injections of EC-SOD transgenic MEF (each at 1.4 x 10(60 cells) on days 28, 35, and 42 after primary immunization. To test EC-SOD activity, blood samples were collected in each group on day 49. The EC-SOD activity was nearly 1.5-fold higher in the transgenic MEF-treated group than in the nontransgenic MEF-treated group (p < 0.05). The severity of arthritis in mice was scored in a double-blind manner, with each paw being assigned a separate clinical score. The severity of arthritis in EC-SOD transgenic MEF-treated mice was significantly suppressed in the arthritic clinical score (p < 0.05). To investigate the alteration of cytokine levels, ELISA was used to measure blood samples. Levels of IL-1beta and TNF-alpha were reduced in the transgenic MEF-treated group (p < 0.05). Abnormalities of the joints were examined by H&E staining. There were no signs of inflammation except for mild hyperplasia of the synovium in the transgenic MEF-treated group. The proliferation of CII-specific T cells was lower in the transgenic MEF-treated mice than in those in the other groups. The transfer of EC-SOD transgenic MEF has shown a therapeutic effect in CIA mice and this approach may be a safer and more effective form of therapy for rheumatoid arthritis.
Cell Transplantation 01/2008; 17(12):1371-80. · 5.13 Impact Factor
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Sang Ryeul Lee,
Jae-Wook Kim,
Bong Soo Kim,
Myoung Ok Kim,
Sung Hyun Kim,
Dong Hun Yoo, Mi-Jung Shin,
Sanggyu Lee,
Young Sik Park,
Yong Bok Park,
Ji Hong Ha,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: In the present study, canine oocytes were exposed to various concentrations of and durations of exposure to EDTA saturated with Ca(2+) (Ca-EDTA), a cell membrane-impermeable metal ion chelator, to determine if parthenogenetic activation could be induced. When oocytes were cultured for 48 or 72 h in parthenogenetic activation medium (PAM) without Ca-EDTA (control) or PAM supplemented with 1 or 5mM Ca-EDTA, the highest rate of pronuclear formation (PN) was obtained in oocytes cultured in 1mM Ca-EDTA for 48 h (8.0%; P<0.05). There was no pronuclear formation in the control group (PAM without Ca-EDTA). Oocytes treated with 5mM Ca-EDTA for 48 h or 1mM Ca-EDTA for 72 h formed a parthenogenetic pronucleus (3.1 and 4.5, respectively). However, there was no pronuclear formation in oocytes treated with 5mM Ca-EDTA for 72 h. In summary, exposure to Ca-EDTA can induce pronuclear formation in canine oocytes.
Theriogenology 04/2007; 67(4):698-703. · 1.96 Impact Factor
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Myoung Ok Kim,
Sung Hyun Kim, Mi Jung Shin,
Dong Beom Lee,
Tae Won Kim,
Kil Soo Kim,
Ji Hong Ha,
Sanggyu Lee,
Yong Bok Park,
Sun Jung Kim,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: We have expressed human erythropoietin (EPO) in transgenic mice using a recombinant EPO cDNA combined with a partial TPO construct. The gene was microinjected using standard techniques and five mice were detected as transgenic by PCR and further used as founders. The life span of the transgenic founders was much shorter than that of their normal littermates. Most of the tissues of the transgenic founders contained human EPO transcripts as judged by RT-PCR. Especially high expression levels were seen in the liver and lung. EPO protein levels in serum were examined by ELISA and ranged from 266, 414 mIU/ml. The number of red blood cell, white blood cell and hemoglobin in the hEPO transgenic mice was higher than in normal mice. These results indicate that overexpression of hEPO is deleterious and can provoke lung failure and erythrocytosis.
Molecules and Cells 03/2007; 23(1):17-22. · 2.18 Impact Factor
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[show abstract]
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ABSTRACT: To determine whether the mammary gland can be used to secrete large quantities of a bioactive heterodimeric protein into milk, we used a bovine beta-casein promoter to target and express human follicle-stimulating hormone (hFSH) in the mammary gland into the milk of transgenic mice. We also identified the effects of hFSH leaked into the bloodstream. Transgenic mice produced a high level (up to 300 mIU/ml) of recombinant hFSH in the mammary gland. Human FSH was expressed in the mammary gland and brain, as determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. In vitro bioactivity was also identified by cyclic AMP (cAMP) assay. The highest activity was showed in the transgenic mice line 11. However, hFSH leaked into the bloodstream was a powerful factor in the generation of breast and ovarian tumors from the transgenic mice line 11. These results suggest that change of endogenous hormones (FSH and progesterone) may affect the morphology and blood cell counts of peripheral blood and, especially, provoke breast and ovarian tumors.
Transgenic Research 03/2007; 16(1):65-75. · 2.75 Impact Factor
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[show abstract]
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ABSTRACT: Human papillomavirus type 16 (HPV16) is a major causative factor in the development of uterine cervical carcinomas. We investigated the role of E6/E7 in tumor formation. Skin-specific E6/E7 transgenic mice showed approximately twice as many tumors compared with nontransgenic mice in dimethylbenz[a]anthracene (DMBA)-initiated and a 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted two-stage skin carcinogenesis. This model showed a significant increase of epidermal cell proliferation in the transgenic mice. The 8-hydroxy-2'deoxyguanosine (8OH-dG) detection assay showed that oxidative DNA damage was significantly higher in the transgenic mice after TPA treatments. The overexpression of E6/E7 in the skin in the DMBA/TPA two-stage-induced carcinogenesis model aggravated the incidence of tumor formation. HPV16 E6/E7 appears to act as an enhancer of carcinogenesis that requires initiation by DMBA and promotion by TPA.
Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics 02/2007; 16(7):325-32. · 1.30 Impact Factor
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Mi Jung Shin,
Jeong-Han Lee,
Dong Hoon Yu,
Bong Soo Kim,
Hei Jung Kim,
Sung-Hyun Kim,
Myoung Ok Kim,
Channy Park,
Byung-Hwa Hyun,
Sanggyu Lee,
Jun-Gyo Suh,
Hong-Seob So,
Raekil Park,
Zae Young Ryoo
[show abstract]
[hide abstract]
ABSTRACT: The spontaneous mutant circling mouse (cir/cir) shows a circling behavior and hearing loss. We produced transgenic mice overexpressing the causative gene, transmembrane inner ear (tmie), for the phenotypic rescue of the circling mouse. Through the continuous breeding with circling mice, the cir/cir homozygous mice carrying the transgene (cir/cir-tg) were produced. The rescued cir/cir -tg mice were able to swim in the water with proper orientation and did not show any circling behavior like wild type mice. Western blot and immunohistochemical analysis exhibited that the transgenic tmie was expressed in the inner ear. Inner and outer hair cells were recovered in the cochlea and spiral ganglion neurons were also recovered in the rescued mice. Auditory brainstem response (ABR) test demonstrated that the cir/cir -tg mice are able to respond to sound. This study demonstrates that tmie transgene can recover the hearing impairment and abnormal behavior in the circling mouse.
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