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1.
Dis Model Mech ; 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38813692

RESUMO

Vertebrate photoreceptors are highly specialized retinal neurons that have cilium-derived membrane organelles called outer segments (OS), which function as platforms for phototransduction. Male germ cell-associated kinase (MAK) is a cilium-associated serine/threonine kinase, and its genetic mutation causes photoreceptor degeneration in mice and retinitis pigmentosa in humans. However, the role of MAK in photoreceptors is not fully understood. Here, we report that zebrafish mak mutants show rapid photoreceptor degeneration during embryonic development. In mak mutants, both cone and rod photoreceptors completely lack OSs and undergo apoptosis. Interestingly, zebrafish mak mutants fail to generate axonemes during photoreceptor ciliogenesis, whereas basal bodies are specified. These data suggest that MAK contributes to axoneme development in zebrafish, in contrast to mouse Mak mutants, which have elongated photoreceptor axonemes. Furthermore, the kinase activity of MAK is critical in ciliary axoneme development and photoreceptor survival. Thus, MAK is required for ciliogenesis and OS formation in zebrafish photoreceptors to ensure intracellular protein transport and photoreceptor survival.

2.
Sci Rep ; 10(1): 17379, 2020 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-33060680

RESUMO

BNip1, which functions as a t-SNARE component of the syntaxin18 complex, is localized on the ER membrane and regulates retrograde transport from Golgi to the ER. BNip1 also has a BH3 domain, which generally releases pro-apoptotic proteins from Bcl2-mediated inhibition. Previously we reported that retinal photoreceptors undergo BNip1-dependent apoptosis in zebrafish ß-snap1 mutants. Here, we investigated physiological roles of BNip1-dependent photoreceptor apoptosis. First, we examined the spatio-temporal profile of photoreceptor apoptosis in ß-snap1 mutants, and found that apoptosis occurs only during a small developmental window, 2-4 days-post-fertilization (dpf), in which an apical photoreceptive membrane structure, called the outer segment (OS), grows rapidly. Transient expression of ß-SNAP1 during this OS growing period prevents photoreceptor apoptosis in ß-snap1 mutants, enabling cone to survive until at least 21 dpf. These observations suggest that BNip1-mediated apoptosis is linked to excessive activation of vesicular transport associated with rapid growth of the OS. Consistently, knockdown of Ift88 and Kif3b, which inhibits protein transport to the OS, rescued photoreceptor apoptosis in ß-snap1 mutants. Treatment with rapamycin, which inhibits protein synthesis via the mTOR pathway, also rescued photoreceptor apoptosis in ß-snap1 mutants. These data suggest that BNip1 performs risk assessment to detect excessive vesicular transport in photoreceptors.


Assuntos
Apoptose/fisiologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células Fotorreceptoras Retinianas Cones/metabolismo , Proteínas de Ligação a Fator Solúvel Sensível a N-Etilmaleimida/fisiologia , Peixe-Zebra/metabolismo , Animais , Células Fotorreceptoras Retinianas Cones/citologia , Peixe-Zebra/embriologia
3.
Pain Pract ; 19(6): 609-620, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30891879

RESUMO

OBJECTIVES: The Multidimensional Pain Readiness to Change Questionnaire 2 (MPRCQ2) is a reliable and valid measure that assesses readiness to adopt a variety of discrete pain self-management responses. We sought to translate and evaluate psychometric properties of the Japanese version of the MPRCQ2 (MPRCQ2-J) in individuals with chronic pain. METHODS: One hundred seventy-three individuals with chronic pain were asked to complete the MPRCQ2-J, as well as measures assessing pain intensity, pain interference, self-efficacy, and general readiness to adopt a self-management approach for pain. Forty-eight of these participants provided additional MPRCQ2-J data to assess test-retest reliability. RESULTS: The findings supported a 2-factor structure of the MPRCQ2-J when error covariances between the some of the nine primary scales were allowed. Adequate internal consistencies of the MPRCQ2-J scales (Cronbach's α ranged 0.71 to 0.86), except for the total score (α = 0.68), were observed. However, adequate test-retest reliabilities (intraclass correlation coefficients ≥ 0.60) were found for only 59% of the MPRCQ2-J scales. The MPRCQ2-J evidenced its construct validity via confirmation of the predicted patterns of associations with validity criterion measures and the anticipated effects of participation in an exercise treatment. DISCUSSION: The findings support the internal consistency (except for the total score) and construct validity for MPRCQ2-J scales. However, potential limitations with respect to test-retest reliability of some of the scales were also suggested. The MPRCQ2-J can be used to examine the role that specific readiness domains of pain self-management responses may play in an adjustment process in Japanese individuals with chronic pain.


Assuntos
Dor Crônica , Medição da Dor/métodos , Psicometria , Inquéritos e Questionários , Adulto , Dor Crônica/psicologia , Dor Crônica/terapia , Comparação Transcultural , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Tradução
4.
Development ; 145(20)2018 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-30322969

RESUMO

In vertebrate lens, lens epithelial cells cover the anterior half of the lens fiber core. Lens epithelial cells proliferate, move posteriorly and start to differentiate into lens fiber cells at the lens equator. Although FGF signaling promotes this equatorial commencement of lens fiber differentiation, the underlying mechanism is not fully understood. Here, we show that lens epithelial cells abnormally enter lens fiber differentiation without passing through the equator in zebrafish vps45 mutants. VPS45 belongs to the Sec1/Munc18-like protein family and promotes endosome trafficking, which differentially modulates signal transduction. Ectopic lens fiber differentiation in vps45 mutants does not depend on FGF, but is mediated through activation of TGFß signaling and inhibition of canonical Wnt signaling. Thus, VPS45 normally suppresses lens fiber differentiation in the anterior region of lens epithelium by modulating TGFß and canonical Wnt signaling pathways. These data indicate a novel role of endosome trafficking to ensure equator-dependent commencement of lens fiber differentiation.


Assuntos
Diferenciação Celular , Endocitose , Cristalino/citologia , Cristalino/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Epitélio/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Integrina beta1/metabolismo , Mutação/genética , Fenótipo , Transporte Proteico , Fator de Crescimento Transformador beta/metabolismo , Proteínas de Transporte Vesicular/genética , Via de Sinalização Wnt , Proteínas de Peixe-Zebra/genética
5.
Sci Rep ; 7: 45962, 2017 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-28378769

RESUMO

Genetic mutations in aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) cause photoreceptor degeneration associated with Leber congenital amaurosis 4 (LCA4) in human patients. Here we report retinal phenotypes of a zebrafish aipl1 mutant, gold rush (gosh). In zebrafish, there are two aipl1 genes, aipl1a and aipl1b, which are expressed mainly in rods and cones, respectively. The gosh mutant gene encodes cone-specific aipl1, aipl1b. Cone photoreceptors undergo progressive degeneration in the gosh mutant, indicating that aipl1b is required for cone survival. Furthermore, the cone-specific subunit of cGMP phosphodiesterase 6 (Pde6c) is markedly decreased in the gosh mutant, and the gosh mutation genetically interacts with zebrafish pde6c mutation eclipse (els). These data suggest that Aipl1 is required for Pde6c stability and function. In addition to Pde6c, we found that zebrafish cone-specific guanylate cyclase, zGc3, is also decreased in the gosh and els mutants. Furthermore, zGc3 knockdown embryos showed a marked reduction in Pde6c. These observations illustrate the interdependence of cGMP metabolism regulators between Aipl1, Pde6c, and Gc3 in photoreceptors.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 6/metabolismo , Guanilato Ciclase/metabolismo , Células Fotorreceptoras Retinianas Cones/citologia , Células Fotorreceptoras Retinianas Cones/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Sobrevivência Celular , GMP Cíclico/metabolismo , Epistasia Genética , Fertilização , Guanilato Ciclase/genética , Mutação/genética , Opsinas/metabolismo , Fenótipo , Estabilidade Proteica , Transporte Proteico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Degeneração Retiniana/patologia , Frações Subcelulares/metabolismo , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
6.
Dev Biol ; 394(1): 94-109, 2014 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-25106852

RESUMO

In the developing retina, neurogenesis and cell differentiation are coupled with cell proliferation. However, molecular mechanisms that coordinate cell proliferation and differentiation are not fully understood. In this study, we found that retinal neurogenesis is severely delayed in the zebrafish stem-loop binding protein (slbp) mutant. SLBP binds to a stem-loop structure at the 3'-end of histone mRNAs, and regulates a replication-dependent synthesis and degradation of histone proteins. Retinal cell proliferation becomes slower in the slbp1 mutant, resulting in cessation of retinal stem cell proliferation. Although retinal stem cells cease proliferation by 2 days postfertilization (dpf) in the slbp mutant, retinal progenitor cells in the central retina continue to proliferate and generate neurons until at least 5dpf. We found that this progenitor proliferation depends on Notch signaling, suggesting that Notch signaling maintains retinal progenitor proliferation when faced with reduced SLBP activity. Thus, SLBP is required for retinal stem cell maintenance. SLBP and Notch signaling are required for retinal progenitor cell proliferation and subsequent neurogenesis. We also show that SLBP1 is required for intraretinal axon pathfinding, probably through morphogenesis of the optic stalk, which expresses attractant cues. Taken together, these data indicate important roles of SLBP in retinal development.


Assuntos
Proteínas de Ligação a RNA/biossíntese , Receptores Notch/metabolismo , Retina/embriologia , Peixe-Zebra/embriologia , Animais , Diferenciação Celular , Proliferação de Células , Quimiocina CXCL12/biossíntese , Proteínas de Fluorescência Verde , Histonas/genética , Mutação , Neurogênese , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genética , Retina/citologia , Transdução de Sinais/genética , Células-Tronco , Ubiquitina-Proteína Ligases/genética , Proteínas de Peixe-Zebra/biossíntese , Proteínas de Peixe-Zebra/genética
7.
Masui ; 62(9): 1127-31, 2013 Sep.
Artigo em Japonês | MEDLINE | ID: mdl-24063142

RESUMO

We report helicopter transportation of a sedated, mechanically ventilated patient with cervical cord injury. A 20-year-old male sustained traumatic injury to the cervical spinal cord during extracurricular activities in a college. On arrival at the hospital, a halo vest was placed on the patient and tracheostomy was performed. On the 38th hospital day, he was transported a distance of 520km by helicopter to a specialized hospital in Fukuoka for medical repatriation. Cabin space was narrow. Since power supply and carrying capacity were limited, battery-driven and portable medical devices were used. In consideration for patient's psychological stress, he was sedated with propofol. RSS (Ramsay sedation scale) scores were recorded to evaluate whether the patient was adequately sedated during helicopter transportation. Prior to transport, we rehearsed the sedation using bispectral index monitoring (BIS) in the hospital to further ensure the patient's safety during the transport.


Assuntos
Resgate Aéreo , Lesões do Pescoço , Respiração Artificial , Traumatismos da Medula Espinal , Transporte de Pacientes/métodos , Traumatismos em Atletas/terapia , Humanos , Hipnóticos e Sedativos/administração & dosagem , Masculino , Lesões do Pescoço/terapia , Propofol/administração & dosagem , Traumatismos da Medula Espinal/terapia , Adulto Jovem
8.
Dev Cell ; 25(4): 374-87, 2013 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-23725763

RESUMO

Intracellular vesicular transport is important for photoreceptor function and maintenance. However, the mechanism underlying photoreceptor degeneration in response to vesicular transport defects is unknown. Here, we report that photoreceptors undergo apoptosis in a zebrafish ß-soluble N-ethylmaleimide-sensitive factor attachment protein (ß-SNAP) mutant. ß-SNAP cooperates with N-ethylmaleimide-sensitive factor to recycle the SNAP receptor (SNARE), a key component of the membrane fusion machinery, by disassembling the cis-SNARE complex generated in the vesicular fusion process. We found that photoreceptor apoptosis in the ß-SNAP mutant was dependent on the BH3-only protein BNip1. BNip1 functions as a component of the syntaxin-18 SNARE complex and regulates retrograde transport from the Golgi to the endoplasmic reticulum. Failure to disassemble the syntaxin-18 cis-SNARE complex caused BNip1-dependent apoptosis. These data suggest that the syntaxin-18 cis-SNARE complex functions as an alarm factor that monitors vesicular fusion competence and that BNip1 transforms vesicular fusion defects into photoreceptor apoptosis.


Assuntos
Apoptose , Fusão de Membrana , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células Fotorreceptoras Retinianas Cones/patologia , Proteínas de Ligação a Fator Solúvel Sensível a N-Etilmaleimida/metabolismo , Animais , Retículo Endoplasmático/metabolismo , Complexo de Golgi/metabolismo , Complexos Multiproteicos/genética , Complexos Multiproteicos/metabolismo , Mutação , Mapeamento de Interação de Proteínas , Estrutura Terciária de Proteína , Transporte Proteico , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Qa-SNARE/genética , Proteínas Qa-SNARE/metabolismo , Células Fotorreceptoras Retinianas Cones/metabolismo , Neurônios Retinianos/metabolismo , Neurônios Retinianos/patologia , Proteínas de Ligação a Fator Solúvel Sensível a N-Etilmaleimida/genética , Peixe-Zebra/genética , Peixe-Zebra/metabolismo
9.
Mech Dev ; 125(11-12): 932-46, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18824093

RESUMO

In mammals, the blockade of the phototransduction cascade causes loss of vision and, in some cases, degeneration of photoreceptors. However, the molecular mechanisms that link phototransduction with photoreceptor degeneration remain to be elucidated. Here, we report that a mutation in the gene encoding a central effector of the phototransduction cascade, cGMP phosphodiesterase 6alpha'-subunit (PDE6alpha'), affects not only the vision but also the survival of cone photoreceptors in zebrafish. We isolated a zebrafish mutant, called eclipse (els), which shows no visual behavior such as optokinetic response (OKR). The cloning of the els mutant gene revealed that a missense mutation occurred in the pde6alpha' gene, resulting in a change in a conserved amino acid. The PDE6 expressed in rod photoreceptors is a heterotetramer comprising two closely related similar hydrolytic alpha and beta subunits and two identical inhibitory gamma subunits, while the PDE6 expressed in cone photoreceptors consists of two homodimers of alpha' subunits, each with gamma subunits. The els mutant displays no visual response to bright light, where cones are active, but shows relatively normal OKR to dim light, where only rods function, suggesting that only the cone-specific phototransduction pathway is disrupted in the els mutant. Furthermore, in the els mutant, cones are selectively eliminated but rods are retained at the adult stage, suggesting that cones undergo a progressive degeneration in the els mutant retinas. Taken together, these data suggest that PDE6alpha' activity is important for the survival of cones in zebrafish.


Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 6/genética , Regulação da Expressão Gênica , Mutação de Sentido Incorreto , Células Fotorreceptoras Retinianas Cones/patologia , Degeneração Retiniana/genética , Alelos , Sequência de Aminoácidos , Animais , Nucleotídeo Cíclico Fosfodiesterase do Tipo 6/fisiologia , Humanos , Transdução de Sinal Luminoso , Dados de Sequência Molecular , Células Fotorreceptoras Retinianas Cones/fisiologia , Células Fotorreceptoras Retinianas Bastonetes/patologia , Células Fotorreceptoras Retinianas Bastonetes/fisiologia , Homologia de Sequência de Aminoácidos , Visão Ocular , Peixe-Zebra
10.
Development ; 132(10): 2273-85, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15829519

RESUMO

In the developing vertebrate hindbrain, the characteristic trajectory of the facial (nVII) motor nerve is generated by caudal migration of the nVII motor neurons. The nVII motor neurons originate in rhombomere (r) 4, and migrate caudally into r6 to form the facial motor nucleus. In this study, using a transgenic zebrafish line that expresses green fluorescent protein (GFP) in the cranial motor neurons, we isolated two novel mutants, designated landlocked (llk) and off-road (ord), which both show highly specific defects in the caudal migration of the nVII motor neurons. We show that the landlocked locus contains the gene scribble1 (scrb1), and that its zygotic expression is required for migration of the nVII motor neurons mainly in a non cell-autonomous manner. Taking advantage of the viability of the llk mutant embryos, we found that maternal expression of scrb1 is required for convergent extension (CE) movements during gastrulation. Furthermore, we show a genetic interaction between scrb1 and trilobite(tri)/strabismus(stbm) in CE. The dual roles of the scrb1 gene in both neuronal migration and CE provide a novel insight into the underlying mechanisms of cell movement in vertebrate development.


Assuntos
Movimento Celular/fisiologia , Nervo Facial/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Neurônios Motores/citologia , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/embriologia , Animais , Sequência de Bases , Movimento Celular/genética , Mapeamento Cromossômico , Cruzamentos Genéticos , Primers do DNA , Gástrula/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Dados de Sequência Molecular , Mutação/genética , Oligonucleotídeos Antissenso , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rombencéfalo/embriologia , Análise de Sequência de DNA
11.
Development ; 130(11): 2479-94, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12702661

RESUMO

The complex, yet highly ordered and predictable, structure of the neural retina is one of the most conserved features of the vertebrate central nervous system. In all vertebrate classes, retinal neurons are organized into laminae with each neuronal class adopting specific morphologies and patterns of connectivity. Using genetic analyses in zebrafish, we demonstrate that N-cadherin (Ncad) has several distinct and crucial functions during the establishment of retinal organization. Although the location of cell division is disorganized in embryos with reduced or no Ncad function, different classes of retinal neurons are generated. However, these neurons fail to organize into correct laminae, most probably owing to compromised adhesion between retinal cells. In addition, amacrine cells exhibit exuberant and misdirected outgrowth of neurites that contributes to severe disorganization of the inner plexiform layer. Retinal ganglion cells also exhibit defects in process outgrowth, with axons exhibiting fasciculation defects and adopting incorrect ipsilateral trajectories. At least some of these defects are likely to be due to a failure to maintain compartment boundaries between eye, optic nerve and brain. Although in vitro studies have implicated Fgf receptors in modulating the axon outgrowth promoting properties of Ncad, most aspects of the Ncad mutant phenotype are not phenocopied by treatments that block Fgf receptor function.


Assuntos
Caderinas/metabolismo , Prosencéfalo/embriologia , Retina/embriologia , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo , Junções Aderentes/metabolismo , Alelos , Animais , Sequência de Bases , Padronização Corporal , Caderinas/genética , Adesão Celular , Divisão Celular , DNA Complementar/genética , Mutação , Neurônios/citologia , Prosencéfalo/metabolismo , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Retina/citologia , Retina/metabolismo , Células Ganglionares da Retina/citologia , Células Ganglionares da Retina/metabolismo , Transdução de Sinais , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
12.
Genesis ; 34(3): 215-20, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12395387

RESUMO

Zebrafish retina contains five morphologically distinct classes of photoreceptors, each expressing a distinct type of opsin gene. Molecular mechanisms underlying specification of opsin expression and differentiation among the cell types are largely unknown. This is partly because mutants affected with expression of a particular class of opsin gene are difficult to find. In this study we established the transgenic lines of zebrafish carrying green fluorescent protein (GFP) gene under the 1.1-kb and 3.7-kb upstream regions of the rod-opsin gene. In transgenic fish, GFP expression initiated and proceeded in the same spatiotemporal pattern with rod-opsin gene. The retinal section from adult transgenic fish showed GFP expression throughout the rod cell layer. These results indicate that the proximal 1.1-kb region is sufficient to drive gene expression in all rod photoreceptor cells. These transgenic fish should facilitate screening of mutants affected specifically with rod-opsin expression or rod cell development by visualization of rod cells by GFP.


Assuntos
Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Células Fotorreceptoras Retinianas Bastonetes/embriologia , Coloração e Rotulagem/métodos , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Regiões 5' não Traduzidas/genética , Animais , Animais Geneticamente Modificados , Imunofluorescência , Proteínas de Fluorescência Verde , Microscopia de Fluorescência , Retina/citologia , Opsinas de Bastonetes/genética
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