[보고서]PLPP/CIN 매개성 ubiquitination에 의한 새로운 신경세포 흥분도 조절 기전 규명

강태천

보고서 정보

주관연구기관

한림대학교
HalLym University

연구책임자

강태천

보고서유형

최종보고서

발행국가

대한민국

언어

한국어

발행년월

2021-03

과제시작연도

2020

주관부처

과학기술정보통신부
Ministry of Science and ICT

연구관리전문기관

한국연구재단
National Research Foundation of Korea

등록번호

TRKO202100016670

과제고유번호

1711109559

사업명

개인기초연구(과기정통부)(R&D)

DB 구축일자

2022-01-22

키워드

피엘피피-씨아이엔.유비퀴티에이션.뇌전증.엔이디디4-2.엠디엠2.PLPP-CIN.Ubiquitination.Epilepsy.NEDD4-2.Mdm2.

표 Effect of PLPP/CIN deletion on NEDD4-2 ubiquitination, and that of NEDD4-2 knockdown on seizure activity in response to KA in PLPP/CIN-/-mice. (A-D) Effect of PLPP/CIN-NEDD4-2 binding on NEDD4-2 ubiquitination following KA injection. (E-H)Effects of NEDD4-2 knockdown on NEDD4-2 expression, and SGK1expression/phosphorylation in response to KAinPLPP/CIN-/-mice. (I-K) Effects of NEDD4-2 knockdown on seizure activity in response to KA in PLPP/CIN-/-mice. (L-M) The effect of NEDD4-2 knockdown on KA-induced neuronal death in PLPP/CIN-/-mice
표 Effects of PLPP/CIN deletion on surface KCNQ and GluA1 expression, GluA1-NEDD4-2 binding, GluA1 ubiquitination and neuronal activity in response to AMPA. (A-C) Effects of PLPP/CIN deletion on surface KCNQ and GluA1 expression following KA injection. (D-G)Effects of PLPP/CIN deletion on GluA1-NEDD4-2 binding and GluA1 ubiquitination. (H-I) Effect of PLPP/CIN deletion on neuronal activity in response to AMPA
표 Effects of PLPP/CIN over-expression on surface KCNQ and GluA1 expression, GluA1-NEDD4-2 bindings and GluA1 ubiquitination. (A-C) Effects of PLPP/CIN over-expression on surface KCNQ and GluA1 expression following KA injection. (D-E) Effects of PLPP/CIN over-expressionon PLPP/CIN-NEDD4-2bindings. (F-G) Effects of PLPP/CIN over-expressionon GluA1-NEDD4-2binding. (H-J) Effects of PLPP/CIN over-expression on NEDD4-2 and GluA1 ubiquitinations
표 Effect of neuronal activity in response to AMPA and the hypothesized role of PCPP/CIN-mediated GluA1 ubiquitination. (A-B) Effects of PLPP/CIN over-expression on neuronal activity in response to AMPA. (C) Scheme of inhibitory role of PLPP/CIN in NEDD4-2-mediated GluA1 ubiquitination. PLPP/CIN dephosphorylates NEDD4-2 S448 site, which leads to self-ubiquitination of NEDD4-2 and subsequently inhibits GluA1 ubiquitination, independent of SGK1 activity
표 Effects of PLPP/CIN deletion and its over-expression onEffects of PLPP/CIN deletion and its over-expression on expressions of AKT, Mdm2 and PSD95, and the phosphorylation levels of AKT and Mdm2 following KA injection
표 The effects of KA on PLPP/CIN-Mdm2, pMdm2-PSD95 and NR2A-PSD95 bindings in vivo
표 Effects of KA on ubiquitinations of Mdm2 and PSD95 (a-c), and that of Mdm2 knockdown on Mdm2, PSD95 and AKT in response to KA in PLPP/CIN-/-mice (d-g)
표 Effects of Mdm2 knockdown on seizure activity in response to KA in PLPP/CIN-/-(KO)mice
표 Scheme of the role of PLPP/CIN in Mdm2-mediated PSD95 ubiquitination. AKT-mediated Mdm2 S166 phosphorylation increases its binding to and ubiquitination of PSD95. Subsequently, PSD95 ubiquitination reduces NR2A-PSD95 bindings, which would facilitate NR2A internalization. PLPP/CIN-mediated Mdm2 dephosphorylation leads to Mdm2 ubiquitination, and abrogates Mdm2-mediated PSD95 degradation in an activity-dependent manner. These PLPP/CIN-mediated functional couplings of Mdm2 and PSD95 exert seizure progression and its seizure severity in response to KA without altering the latency of seizure on-set
표 PLPP/CIN-mediated NF2 dephosphorylation. (a-e) In vitro assay using recombinant proteins. (a) Representative western blot data demonstrating PLPP/CIN-mediated NF2 dephosphorylation. (b-c) Quantification of NF2-S10 (b) and -518 (c) phosphorylation levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (*p < 0.05 vs. the presence of PLPP/CIN and PKAc; n = 7). (d) Representative co-immunoprecipitation data demonstrating PLPP/CIN-NF2 and -PKAc bindings. (e) Quantification of PLPP/CIN-NF2 and -PKAc bindings. Open circles indicate each individual value. Horizontal bars indicate mean value (*p < 0.05 vs. the presence of PLPP/CIN and PKAc; n = 7). (f-g) Effects of KA on PLPP/CIN-NF2 and -PKAc bindings in WT and PLPP/CINTg mice in vivo. (f) Representative co-immunoprecipitation data demonstrating PLPP/CIN-PKAc and NF2 bindings. (g) Quantification of PLPP/CIN-PKAc and -NF2 bindings. Open circles indicate each individual value. Horizontal bars indicate mean value (*,#p < 0.05 vs. WT and control animals, respectively; n = 7)
표 PLPP/CIN-mediated NF2 dephosphorylation. (a-e) In vitro assay using recombinant proteins. (a) Representative western blot data demonstrating PLPP/CIN-mediated NF2 dephosphorylation. (b-c) Quantification of NF2-S10 (b) and -518 (c) phosphorylation levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (*p < 0.05 vs. the presence of PLPP/CIN and PKAc; n = 7). (d) Representative co-immunoprecipitation data demonstrating PLPP/CIN-NF2 and -PKAc bindings. (e) Quantification of PLPP/CIN-NF2 and -PKAc bindings. Open circles indicate each individual value. Horizontal bars indicate mean value (*p < 0.05 vs. the presence of PLPP/CIN and PKAc; n = 7). (f-g) Effects of KA on PLPP/CIN-NF2 and -PKAc bindings in WT and PLPP/CINTg mice in vivo. (f) Representative co-immunoprecipitation data demonstrating PLPP/CIN-PKAc and NF2 bindings. (g) Quantification of PLPP/CIN-PKAc and -NF2 bindings. Open circles indicate each individual value. Horizontal bars indicate mean value (*,#p < 0.05 vs. WT and control animals, respectively; n = 7)
표 Effects of PLPP/CIN deletion on seizure activity, NF2 phosphorylation and PLPP/CIN-NF2 binding in response to KA. (a) Representative EEG traces and frequency-power spectral temporal maps in response to KA. (b) Quantification of total EEG power (seizure intensity ) in response to KA (#p < 0.05 vs WT animals; n = 7, respectively ). (c) Repersentative western blots of PLPP/CIN, NF2, pNF2-S10, pNF2-S518, PKAc and pPKAc in WT and and PLPP/CINTg mice in vivo. (d-h) Quantification of MF2, pNF2-S10, pNF2-S518, PKAc and pPKAc levels. Open circles indicate each indiviual value. Horizontal bars indicate mean value (*#p < 0.05 vs. WT and control animals, respectively; n = 7). (i-j) Effects of KA on PLPP/CIN-NF2 and -PKAc bindings in WT and PLPP/CIN-/- mice in vivo. (i) Representative co -immunoprecipitation data demonstrating PLPP/CIN-PKAc and MF2 bindings. (j) Quantification of PLPP/CIN-PKAc and -NF2 bindings. Open circles indicatd each individual value. Horizontal bars indicate mean value ( *#p < 0.05 vs. WT and control animals, respectively; n = 7)
표 Effects of NF2 knockdown on NF2 and cofilin phosphorylations in PLPP/CINTg and PLPP/CIN-/- mice under physiological condition. (a) Representative western blots of NF2, pNF2-S10, pNF2-S518, cofilin, pCofilin-S3 and F-actin. (b-k) Quantification of NF2, pNF2-S10, pNF2-S518, cofilin, pCofilin-S3 and F-actin levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (*,#p < 0.05 vs. WT and control siRNA-treated animals, respectively; n = 7)
표 Linear regression analysis between NF2 expression and F-actin contents and the effects of NF2 knockdown on seizure activity in response to KA in PLPP/CINTg and PLPP/CIN-/- mice. (a) Linear regression analysis between NF2 expression and F-actin contents within control siRNA- and NF2 siRNA-treated groups of WT, PLPP/CINTg and PLPP/CIN-/- mice under physiological conditions. (b-g) Representative EEG traces and frequency-power spectral temporal maps (b, d and f) and quantification of total EEG power (seizure intensity) in response to KA (c, e and g) in WT (b-c), PLPP/CINTgPLPP/CIN-/- mice (#p < 0.05 vs. control siRNA-treated animals; n = 7, respectively)
표 Effects of NF2 knockdown on protein and phosphorylation levels of NF2, Mdm2 and PSD95 in PLPP/CINTg mice following KA injection. (a) Representative western blots of NF2, pNF2-S10, pNF2-S518, Mdm2, pMdm2-S166 and PSD95. (b-g) Quantification of NF2, pNF2-S10, pNF2-S518, Mdm2, pMdm2-S166 and PSD95 levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (*,#,$p < 0.05 vs. WT, control siRNA- and control (non-KA-treated) animals, respectively; n = 7)
표 Effects of NF2 knockdown on protein and phosphorylation levels of NF2, Mdm2 and PSD95 inPLPP/CIN-/-mice following KA injection, and linear regression analysis between NF2 and Mdm2/PSD95 levels. (a) Representative western blots of NF2, pNF2-S10, pNF2-S518, Mdm2, pMdm2-S166 and PSD95. (b-g) Quantification of NF2, pNF2-S10, pNF2-S518, Mdm2, pMdm2-S166 and PSD95 levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (*,#,$p < 0.05 vs. WT, control siRNAand control (non-KA-treated) animals, respectively; n = 7). (h-i) Linear regression analysis between NF2 and Mdm2/PSD95 levels within control siRNA- and NF2 siRNA-treated groups of WT and PLPP/CINTg mice following KA injection
표 Effects of NF2 knockdown on CA3 neuronal death in WT, PLPP/CINTg and PLPP/CIN-/ mice following KA injection and the role of PLPP/CIN inn NF2=S10 and Mdm2-S116 dephosphoylation. (a) Representative photos of FJB positive degenerating neurons in the CA1 region. (b) Quantification of the number of FJB positive neurons in response to KA. Open circles indicate each individual value. Horizontal bars indicate mean ( *#p < 0.05 vs. WT and control siRNA-treated animal; n = 7, respectively). C) Scheme of the role of PLPP/CIN in NF2-S10 and Mdm2-S166 dephosphorylation PLPP/CIN -mediated NF2 dephosphorylation decreases F-actin stability and increases NMDAR-PSD95 co-assembly by eliminating the local F-actin barrier. PLPP/CIN also inhibits Mdm2 activity as an E3 ubiquitin ligase for PSD95 by enhancing its S166 dephosphorylation and NF2-mediated Mdm2 degradation

과제명(ProjectTitle) :	-
연구책임자(Manager) :	-
과제기간(DetailSeriesProject) :	-
총연구비 (DetailSeriesProject) :	-
키워드(keyword) :	-
과제수행기간(LeadAgency) :	-
연구목표(Goal) :	-
연구내용(Abstract) :	-
기대효과(Effect) :	-

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 제목(한글), 저자명(한글), 발행일자, 전자원문, 초록(한글), 초록(영문) 관리번호, 제목(한글), 제목(영문), 저자명(한글), 저자명(영문), 주관연구기관(한글), 주관연구기관(영문), 발행일자, 총페이지수, 주관부처명, 과제시작일, 보고서번호, 과제종료일, 주제분류, 키워드(한글), 전자원문, 키워드(영문), 입수제어번호, 초록(한글), 초록(영문), 목차
저장형식	Text(ASCII format) Excel format
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

[국가R&D연구보고서] PLPP/CIN 매개성 ubiquitination에 의한 새로운 신경세포 흥분도 조절 기전 규명
Novel mechanism of neuronal excitability by PLPP/CIN-mediated ubiquitination 원문보기

초록 ▼

표/그림 (17)

표/그림 (17)

참고문헌 (25)

연구과제 타임라인

활용도 분석정보

활용도 Top5 보고서

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

연관된 기능

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

연합인증

[국가R&D연구보고서] PLPP/CIN 매개성 ubiquitination에 의한 새로운 신경세포 흥분도 조절 기전 규명 Novel mechanism of neuronal excitability by PLPP/CIN-mediated ubiquitination 원문보기

초록 ▼

표/그림 (17) 모든 표/그림 보기

표/그림 (17) 슬라이드로 보기

참고문헌 (25)

연구과제 타임라인

전체(0) 논문(0) 특허(0) 보고서(0)

전체(0) 논문(0) 특허(0) 보고서(0)

활용도 분석정보

활용도 Top5 보고서 더보기

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

연관된 기능

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

[국가R&D연구보고서] PLPP/CIN 매개성 ubiquitination에 의한 새로운 신경세포 흥분도 조절 기전 규명
Novel mechanism of neuronal excitability by PLPP/CIN-mediated ubiquitination 원문보기

표/그림 (17)

표/그림 (17)

활용도 Top5 보고서