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신경세포 시냅스에서 Shank2, Homer 1b와 PLC-β3의 신호전달 복합체 형성
황종익,신금주,류성호,서판길 한국뇌학회 2001 한국뇌학회지 Vol.1 No.2
포스포리파아제 C-베타(phospholipase C-β: PLC-β) 동위효소는 G-proteins-coupled receptors(GPCR)와 그의 신호전달 매개체인 heterotrimeric G proteins에 의해 활성화된다. PLC-β는 효소적 활성에 필요한 영역 외에 카르복시 말단에 PSD-95/Dlg/ZO-1 (PDZ)-binding motif를 가진다. 이 motif는 PLC-β가 PDZ domain을 가진 단백질들과 결합함으로써 GPCR 매개 신호전달에서 특이적인 역할을 수행할 가능성을 제시한다. 이에 본 연구자들은 효모 two-hybrid assay를 통하여 Shank2가 PLC-β3와 결합함을 이전에 확인하였다. 본 연구에서는, 면역침전반응을 통해 두 단백질의 결합은 PLC-β3의 카르복시 말단의 PDZ-binding motif를 통하여 이루어짐을 확인하였고 신경세포의 시냅스에서 mGluR1 및 IP3 수용체와 결합하는 Homer 1b 와 Shank2가 PLC-β3와 함께 위치하는 것을 발견하였다. 이러한 결과는 전자현미경을 이용하여 Homer 1b와 Shank2가 위치하는 PSD 영역에서 PLC-β3가 존재함을 확인하고 mGluR1과 Shank2, Homer 1b, PLC-β3가 해마와 소뇌의 많은 부분에서 동시에 발현되는 것을 확인함으로써 다시 한번 입증되었다. 결과를 종합해 볼 때, 이들 세 단백질은 같은 신경세포의 시냅스에서 발현되어 복합체를 형성함으로써 mGluR1에 의해 매개되는 신호전달을 보다 효율적으로 수행할 수 있을 것으로 예상된다. Phospholipase C-β isotypes activated by G protein-coupled receptors (GPCR) and heterotrimeric G proteins carry a PSD-95/Dlg/ZO-1 (PDZ)-binding motif in their carboxyl terminus. This motif may enable PLC-β isotypes to play specific roles in GPCR signaling through interacting with PDZ-containing proteins. We identified Shank2 as a PLC-β3 binding protein, using yeast two-hybrid system in the previous study, and here we characterized the binding of PLC-β3 with Shank2. Immunoprecipitation study showed that Shank2 interacts with PDZ-binding motif of PLC-β3. Moreover, PLC-β3 was colocalized with Shank2 and Homer 1b that interacts with mGluR1 and IP3 receptor on the synapse of primary-cultured neuronal cells. Immunogold EM also showed that PLC-β3 immunoreactivity was detected at the PSD of the CA1 pyramidal neurons. In several regions of hippocampus and cerebellum, the expression pattern of Shank2 was similar to that of mGluR1, Homer 1b, and PLC-β3. These results suggest that PLC-β3, Shank2, and Homer 1b may act as active intermediates in mGluR1-mediated signal transduction by forming signaling complex in neuronal synapses.
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2,2′,4,6,6′-Pentachlorobiphenyl Induces Mitotic Arrest and p53 Activation
Shin, Kum-Joo,Kim, Sun-Hee,Kim, Dohan,Kim, Yun-Hee,Lee, Han-Woong,Chang, Yoon-Seok,Gu, Man-Bock,Ryu, Sung Ho,Suh, Pann-Ghill Oxford University Press 2004 TOXICOLOGICAL SCIENCES Vol.78 No.2
<P>Polychlorinated biphenyls (PCBs), a class of persistent organic pollutants (POPs), have been considered to be involved in cancers, but the underlying mechanisms are not known well. Various cancers are closely related to genetic alteration; therefore, we investigated the effect of PCBs on genetic stability, through p53, a guardian of genome, in NIH 3T3 fibroblasts. Among several congeners examined, 2,2′,4,6,6′-pentachlorobiphenyl (PeCB) specifically activated p53-dependent transcription. It also induced p53 nuclear accumulation, but did not cause DNA strand breakage. On the other hand, cell cycle progression that is closely connected to p53 was affected by 2,2′,4,6,6′-PeCB, resulting in mitotic arrest. In the arrested cells, mitotic spindle damage was detected. Moreover, in the absence of functional p53, polyploidy was caused by 2,2′,4,6,6′-PeCB. These results imply that 2,2′,4,6,6′-PeCB induces mitotic arrest by interfering with mitotic spindle assembly, followed by genetic instability which triggers p53-activating signals to prevent further polyploidization. Taking these findings together, we suggest that 2,2′,4,6,6′-PeCB could be involved in cancer development by causing genetic instability through mitotic spindle damage, which brings about aneuploidy in p53-deficient tumor cells.</P>
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Joo Young Shin,Jin Seok Choi,Joo Youn Oh,Mee Kum Kim,Jin Hak Lee,Won Ryang Wee 대한안과학회 2010 Korean Journal of Ophthalmology Vol.24 No.3
Purpose: To evaluate corneal biomechanical properties in eyes that had previously undergone penetrating keratoplasty(PK) using the ocular response analyzer (ORA). Methods: We recruited 26 patients who had received unilateral PK. Corneal hysteresis (CH), corneal resistance factor (CRF), Goldmann-correlated intraocular pressure (IOPg), and cornea-compensated intraocular pressure (IOPcc) were measured with the ORA and were compared to the measurements from the contralateral eyes that did not undergo PK. Results: The CH was 8.95±2.59 mmHg in eyes that underwent PK and 9.78±1.45 mmHg in the contralateral eyes that did not undergo PK (p=0.077). The CRF was 10.26±2.64 mmHg in post-PK eyes and 9.75±1.45 mmHg in the contralateral eyes (p=0.509), and the CH-CRF was significantly smaller in post-PK eyes (-1.31±2.32 mmHg in post-PK eyes vs. 0.03±0.88 mmHg in fellow eyes, p=0.016). The IOPg and IOPcc were significantly higher in the PK group than they were in the control group. The IOPcc’s were 20.81±7.81 mmHg and 16.27±2.49 mmHg in post-PK and control eyes, respectively (p=0.011); and the IOPg’s were 19.22±7.34 mmHg and 15.07±3.03 mmHg in post-PK and control eyes, respectively (p=0.019). The IOPcc-g’s were 1.59±2.81 mmHg and 1.21±1.30 mmHg in post-PK and control eyes, respectively (p=0.412), and the central corneal thickness (CCT)’s were 489.11±90.60 μm and 556.24±42.84 μm in post-PK and control eyes, respectively (p=0.068). Conclusions: Following PK, CH tended to decrease while CRF tended to increase, significantly decreasing CH-CRF. A significantly higher intraocular pressure and a thinner CCT following PK may have contributed to the observed changes in these corneal biomechanical parameters.
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( Young Joo Shin ),( Mee Kum Kim ),( Joo Youn Oh ),( Won Ryang Wee ),( Jin Hak Lee ),( Jung Hwa Ko ),( Hyun Ju Lee ),( Jae Lim Lee ),( Byung Moo Min ),( Young Suk Sohn ) 대한안과학회 2008 Korean Journal of Ophthalmology Vol.22 No.2
Purpose: To investigate the short-term efficacy of topical immunosuppressive agents on the survival of cultivated allo-conjunctival equivalents. Methods: Twenty-five eyes of New Zealand white rabbits were included. Temporal conjunctivae were trephined to a diameter of 7.5 mm, and then cultured allo-conjunctival epithelial cells on amniotic membrane were transplanted onto them. Various immunosuppressants including steroid, cyclosporine, and rapamycin were applied topically four times a day for a week. Epithelial defects and graft edema were graded daily. Numbers of inflammatory cells were measured in H&E. PKH26 and cytokeratin 4 and 7 were immunostained. Results: Earlier epithelialization was observed in 1% steroid-treated eyes and defects persisted significantly in 0.5% CsA applied eyes. In histology, PKH26 positive cells considered as donor cells were only found in 1% steroid or 0.01% rapamycin applied eyes. 1% steroid- or 0.01% rapamycin-applied eyes both showed positive staining for keratin-4 and -7. Inflammatory cells were less found in 1% steroid or 0.01% rapamycin treated eyes. Conclusions: Topical steroid or rapamycin can help to suppress acute inflammation and enhance the acute survival of transplanted conjunctival cells.
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Young Joo Shin,Mee Kum Kim,Joo Youn Oh,Won Ryang Wee,Jin Hak Lee,Jung Hwa Ko,Hyun Ju Lee,Jae Lim Lee,Byung Moo Min,Young Suk Sohn 대한안과학회 2008 Korean Journal of Ophthalmology Vol.22 No.2
Purpose: To investigate the short‐term efficacy of topical immunosuppressive agents on the survival of cultivated allo‐conjunctival equivalents. Methods: Twenty‐five eyes of New Zealand white rabbits were included. Temporal conjunctivae were trephined to a diameter of 7.5 mm, and then cultured allo‐conjunctival epithelial cells on amniotic membrane were transplanted onto them. Various immunosuppressants including steroid, cyclosporine, and rapamycin were applied topically four times a day for a week. Epithelial defects and graft edema were graded daily. Numbers of inflammatory cells were measured in H&E. PKH26 and cytokeratin 4 and 7 were immunostained. Results: Earlier epithelialization was observed in 1% steroid‐treated eyes and defects persisted significantly in 0.5% CsA applied eyes. In histology, PKH26 positive cells considered as donor cells were only found in 1% steroid or 0.01% rapamycin applied eyes. 1% steroid‐ or 0.01% rapamycin‐applied eyes both showed positive staining for keratin‐4 and ‐7. Inflammatory cells were less found in 1% steroid or 0.01% rapamycin treated eyes. Conclusions: Topical steroid or rapamycin can help to suppress acute inflammation and enhance the acute survival of transplanted conjunctival cells. Korean Journal of Ophthalmology 22(2):123-129, 2008
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Shin, Jieun,Kim, Jong-Eun,Pak, Kum-Ju,Kang, Jung Il,Kim, Tae-Seok,Lee, Sang-Yoon,Yeo, Ik-Hyun,Park, Jung Han Yoon,Kim, Jong Hun,Kang, Nam Joo,Lee, Ki Won,Cifuentes, Alejandro MDPI 2017 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.18 No.3
<P>Soybean-derived isoflavones have been investigated for their preventative effects against UV-induced symptoms of skin damage including wrinkle formation and inflammation. <I>Haematococcus pluvialis</I> is a freshwater species of Chlorophyta that contains high concentrations of the natural carotenoid pigment astaxanthin. Astaxanthin is known to be involved in retinoic acid receptor (RAR) signaling and previously been associated with the inhibition of activator protein (AP)-1 dependent transcription. Based on previous studies, we hypothesized that a combination of soy extract (SE) and <I>Haematococcus</I> extract (HE) may prevent UVB-induced photoaging through specific signaling pathways, as measured by UVB-induced wrinkling on hairless mice skin and expression changes in human dermal fibroblasts (HDFs). The 1:2 ratio of SE and HE mixture (SHM) showed the optimal benefit in vivo. SHM was found to inhibit wrinkle formation via the downregulation of matrix metalloproteinase (MMP)-1 mRNA and protein expression. SHM also inhibited mitogen-activated protein kinase (MAPK) phosphorylation and the transactivation of AP-1 which plays an important role in regulating MMP expression. These results highlight the potential for SHM to be developed as a therapeutic agent to prevent UVB-induced skin wrinkling.</P>
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( Joo Youn Oh ),( Mee Kum Kim ),( Kyung Seon Shin ),( Won Ryang Wee ),( Jin Hak Lee ) 한국조직공학·재생의학회 2010 조직공학과 재생의학 Vol.7 No.1
Purpose: To investigate the expression of stem cell-associated and corneal differentiation markers in human bone marrow-derived MSCs (hMSCs) and to examine the influence of cell culture conditions on the potential of hMSCs to differentiate into corneal epithelial cells. Methods: We identified and characterized stem cell-associated and corneal differentiation markers in hMSCs using RT PCR and immunocytochemistry, and compared it to the marker expression seen in human corneal limbal epithelial cells (hLECs). We also examined the influence of cell culture conditions, such as serial passage, cell plating density, presence of feeder cells, and addition of epidermal (EGF) and fibroblast growth factors (FGF) at varying densities, on the marker expression of hMSCs. Results: Both hMSCs and hLECs expressed Cnx43. However, hMSCs did not express p63, ABCG2, OCT4, K12, or MUC16, which were expressed in hLECs. MUC1 was highly expressed in hMSCs, while it was not expressed in hLECs. Notably, hMSCs lost the ability to express MUC1, when 20 ng/ml of EGF was added to the culture in the presence of NIH/3T3 feeder cells. However, the culture duration, cell plating density, and use of feeder cells did not exert a significant effect on the marker expression in hMSCs. Conclusion: P63, K12, and MUC16 can be used as putative positive markers and MUC1 as a putative negative marker for differentiation of hMSCs to hLECs. Differentiation of hMSCs into corneal epithelial-like phenotype was not induced by varying the culture duration or plating density or by using NIH/3T3 fibroblasts as a feeder.
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