Stimulation-induced changes in diffusion and structure of calmodulin and calmodulin-dependent protein kinase II proteins in neurons

Heidarinejad, Morteza; Nakamura, Hideki; Inoue, Takafumi

doi:10.1016/j.neures.2018.01.003

Cited by 5 publications

(3 citation statements)

References 115 publications

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“…Interestingly C2C was well distributed along sensory neuron axons that innervated muscle tissue. This may indicate the C2I protein undergoes intracellular diffusion as has been observed with other neuron targeting proteins 56 and it suggests that C2C could influence calcium channel activity at locations distal to its site of injection. Alternatively, the apparent distribution of C2C is also consistent with GT1b receptor location along sensory neuron axons in mouse tissue.…”

Section: Discussionmentioning

confidence: 71%

Intracellular G-actin targeting of peripheral sensory neurons by the multifunctional engineered protein C2C confers relief from inflammatory pain

Allen

Zhou

Wilhelm

et al. 2020

Sci Rep

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The engineered multifunctional protein C2C was tested for control of sensory neuron activity by targeted G-actin modification. C2C consists of the heptameric oligomer, C2II-CI, and the monomeric ribosylase, C2I. C2C treatment of sensory neurons and SH-SY5Y cells in vitro remodeled actin and reduced calcium influx in a reversible manner. C2C prepared using fluorescently labeled C2I showed selective in vitro C2I delivery to primary sensory neurons but not motor neurons. Delivery was dependent on presence of both C2C subunits and blocked by receptor competition. Immunohistochemistry of mice treated subcutaneously with C2C showed colocalization of subunit C2I with CGRP-positive sensory neurons and fibers but not with ChAT-positive motor neurons and fibers. The significance of sensory neuron targeting was pursued subsequently by testing C2C activity in the formalin inflammatory mouse pain model. Subcutaneous C2C administration reduced pain-like behaviors by 90% relative to untreated controls 6 h post treatment and similarly to the opioid buprenorphene. C2C effects were dose dependent, equally potent in female and male animals and did not change gross motor function. One dose was effective in 2 h and lasted 1 week. Administration of C2I without C2II-CI did not reduce pain-like behavior indicating its intracellular delivery was required for behavioral effect.

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Section: Discussionmentioning

confidence: 71%

Intracellular G-actin targeting of peripheral sensory neurons by the multifunctional engineered protein C2C confers relief from inflammatory pain

Allen

Zhou

Wilhelm

et al. 2020

Sci Rep

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“…previous intracellular FCS studies (19,(43)(44)(45) showing much smaller D values in intracellular FCS recordings than those obtained in the cell-free FCS.…”

Section: Quantification Of Mrna Dynamics By Fcs and Retf Probementioning

confidence: 66%

Quantification of native mRNA dynamics in living neurons using fluorescence correlation spectroscopy and reduction-triggered fluorescent probes

Fujita

Oikawa

Hayakawa

et al. 2020

Journal of Biological Chemistry

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RNA localization in subcellular compartments is essential for spatial and temporal regulation of protein expression in neurons. Several techniques have been developed to visualize mRNAs inside cells, but the study of the behavior of endogenous and nonengineered mRNAs in living neurons has just started. In this study, we combined reduction-triggered fluorescent (RETF) probes and fluorescence correlation spectroscopy (FCS) to investigate the diffusion properties of activity-regulated cytoskeleton-associated protein (Arc) and inositol 1,4,5-trisphosphate receptor type 1 (Ip3r1) mRNAs. This approach enabled us to discriminate between RNA-bound and unbound fluorescent probes and to quantify mRNA diffusion parameters and concentrations in living rat primary hippocampal neurons. Specifically, we detected the induction of Arc mRNA production after neuronal activation in real time. Results from computer simulations with mRNA diffusion coefficients obtained in these analyses supported the idea that free diffusion is incapable of transporting mRNA of sizes close to those of Arc or Ip3r1 to distal dendrites. In conclusion, the combined RETF-FCS approach reported here enables analyses of the dynamics of endogenous, unmodified mRNAs in living neurons, affording a glimpse into the intracellular dynamics of RNA in live cells.

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“…En mamíferos esta proteína está implicada en la interacción y activación de varias fosfatasas como la MAPK fosfatasa I (Ishida et al, 2009) o la calcineurina (Chyan et al, 2017;Klee et al, 1979) y de varias quinasas como la quinasa dependiente de calcio/calmodulina II (CaMKII), implicada en plasticidad sináptica junto con la calcineurina (Heidarinejad et al, 2018). La calmodulina también regula la actividad de otras proteínas señalizadoras como los receptores de estrógenos, la glutamato descarboxilasa o diversos canales de Ca ++ o K + en las neuronas (Villalobo et al, 2018).…”

Section: La Calmodulina Como Proteína Reguladora Y Adaptadoraunclassified

Función de la fosfatasa calcineurina en la citocinesis de Schizosaccharomyces pombe

Arribas¹

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CDRE: Elementos de respuesta a calcineurina (Calcineurin-Dependent Response Element) CDK: quinasa dependiente de ciclina DIC: Contraste interdiferencial (Differential Interphase Contrast) dNTPs: Desoxirribonucleotido trifosfato D.O.: Densidad óptica DTT: Ditiotreitol EDTA: Ácido etilén diamino tetra-acético EGTA: Ácido etilén glicol bis tetra-acético EMM: Medio mínimo para S. pombe (Edimburgh Minimal Media) FK506: Tacrólimus FASP: Preparación de la muestra mediante filtro (Filter-aided sample preparation) F-BAR: Fes/CIP4 homology-BAR GAP: Proteína activadora de la actividad GTPásica (GTPase Activating Protein) GBP: Proteína de union a GFP (GFP Binding Protein) GDP: Guanosina 5'-difostato GEF: Factor intercambiador de nucleótidos de guanina (Guanine Nucleotide Exchange Factor) GFP: Proteína verde fluorescente (Green Fluorescent Protein) GST: Glutation S transferasa Abreviaturas XVI GTP: Guanosina 5'-trifosfato h: Horas HA: Epítopo procedente de la hemaglutinina del virus humano de la gripe (Hemagglutinin Antigen) HRP: Peroxidasa de rábano (Horseradish Peroxidase) iMTOC: Centro organizador de microtúbulos de interfase (Interphase Microtubule Organizer Centre) IP: Inmunoprecipitado IPTG: Isopropil-β-D-tiogalactopiranósido KDa: Kilodalton kb: Kilobase LB: Medio Luria-Bertani para E. coli LC-MS/MS: Cromatografía líquida-espectrometría de masas/espectrometría de masas (Liquid chromatography-mass spectrometry/mass spectrometry) LIM: Dominio proteico cuyas iniciales proceden de las proteinas Lin11, Isl-1 y Mec-3 MBC: Metil 2-Benzimidazol Carbamato MEA: Medio agar extracto de malta (Malt Extract Agar) min: Minutos MTOC: Centro Organizador de Mirotubulos (Microtubule Organizer Center) NETO: Comienzo del crecimiento por el polo celular nuevo (New End Take Off) nmt: Promotor inhibido por tiamina (No Messenger with Thiamine) NP-40: Nonidet P-40 OE: Sobreexpresion de un gen (OverExpression) OETO: Comienzo del crecimiento por el polo celular antiguo (Old End Take Off) ORF: Fase de lectura abierta (Open Reading Frame) pb: Pares de bases PBS: Tampón fosfato salino (Phosphate-Buffered Saline) Abreviaturas XVII PCR: Reacción en cadena de la polimerasa (Polymerase Chain Reaction) PMSF: Fluoruro de ortometil fenil sulfonilo PVDF: Fluoruro de polivinilideno RBD: Dominio de unión a RhoA (RhoA Binding Domain) RFP: Proteína roja fluorescente (Red Fluorescent Protein) Rho: Ras Homology rpm: Revoluciones por minuto SDS: Dodecil sulfato sódico SDS-PAGE: Electroforesis en gel de poliacrilamida en condiciones desnaturalizantes (PolyAcrilamide Gel Electrophoresis) SH3: Dominio con homologia a Src3 (Src3 Homology)

show abstract

Stimulation-induced changes in diffusion and structure of calmodulin and calmodulin-dependent protein kinase II proteins in neurons

Cited by 5 publications

References 115 publications

Intracellular G-actin targeting of peripheral sensory neurons by the multifunctional engineered protein C2C confers relief from inflammatory pain

Intracellular G-actin targeting of peripheral sensory neurons by the multifunctional engineered protein C2C confers relief from inflammatory pain

Quantification of native mRNA dynamics in living neurons using fluorescence correlation spectroscopy and reduction-triggered fluorescent probes

Función de la fosfatasa calcineurina en la citocinesis de Schizosaccharomyces pombe

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