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1.
Science ; 373(6553)2021 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-34437090

RESUMO

The ability to perceive and respond to environmental stimuli emerges in the absence of sensory experience. Spontaneous retinal activity prior to eye opening guides the refinement of retinotopy and eye-specific segregation in mammals, but its role in the development of higher-order visual response properties remains unclear. Here, we describe a transient window in neonatal mouse development during which the spatial propagation of spontaneous retinal waves resembles the optic flow pattern generated by forward self-motion. We show that wave directionality requires the same circuit components that form the adult direction-selective retinal circuit and that chronic disruption of wave directionality alters the development of direction-selective responses of superior colliculus neurons. These data demonstrate how the developing visual system patterns spontaneous activity to simulate ethologically relevant features of the external world and thereby instruct self-organization.


Assuntos
Fluxo Óptico , Retina/fisiologia , Células Ganglionares da Retina/fisiologia , Visão Ocular/fisiologia , Vias Visuais , Potenciais de Ação , Células Amácrinas/fisiologia , Animais , Animais Recém-Nascidos , Axônios/fisiologia , Proteínas do Citoesqueleto/genética , Camundongos , Movimento (Física) , Mutação , Piridazinas/farmacologia , Receptores de GABA-A/metabolismo , Retina/crescimento & desenvolvimento , Análise Espaço-Temporal , Colículos Superiores/fisiologia
2.
Int J Mol Sci ; 22(16)2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34445064

RESUMO

Photoreceptors are critical components of the retina and play a role in the first step of the conversion of light to electric signals. With the discovery of the intrinsically photosensitive retinal ganglion cells, which regulate non-image-forming visual processes, our knowledge of the photosensitive cell family in the retina has deepened. Photoreceptor development is regulated by specific genes and proteins and involves a series of molecular processes including DNA transcription, post-transcriptional modification, protein translation, and post-translational modification. Single-cell sequencing is a promising technology for the study of photoreceptor development. This review presents an overview of the types of human photoreceptors, summarizes recent discoveries in the regulatory mechanisms underlying their development at single-cell resolution, and outlines the prospects in this field.


Assuntos
Células Fotorreceptoras de Vertebrados/citologia , Retina/crescimento & desenvolvimento , Análise de Célula Única/métodos , Animais , Humanos , Organoides/citologia , Organoides/embriologia , Organoides/crescimento & desenvolvimento , Células Fotorreceptoras de Vertebrados/metabolismo , Retina/citologia , Retina/embriologia
3.
Zoolog Sci ; 38(4): 326-331, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34342953

RESUMO

The adult lamprey retina has two types of photoreceptor cells, short and long photoreceptor cells, which are equivalent to rods and cones of other vertebrates. In contrast, the retina of lamprey larvae only contains a single type of photoreceptor cell, which appears to correspond to the short photoreceptor cell. However, the developmental pattern of the long photoreceptor cell is unknown. Previously, we reported that antibodies against rhodopsin and iodopsin (the chicken red cone opsin) could discriminate between the outer segments of short and long photoreceptor cells, respectively, in the retina of adult Japanese river lamprey (Lethenteron camtschaticum). Here, we immunohistochemically investigate the appearance of long photoreceptor cells in the larval and adult retinas of the Far Eastern brook lamprey (Lethenteron reissneri), which is a close relative of the Japanese river lamprey, by using anti-iodopsin antibody. We found that iodopsin immunoreactivity was localized not only in the adult retina but also in the larval retina. Moreover, we examined the immunohistochemical localization of signal transduction molecules, such as transducin and arrestin, in the iodopsin-immunoreactive cells of the larval retina. The iodopsin-immunoreactive cells also contained both transducin and arrestin, suggesting that long photoreceptor cells are already functional in the larval stage before the acquisition of visual function. Our results suggest that the iodopsin-immunoreactive cells may be related to not only cone vision in the adult but also photoreception in the larval lamprey.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Imuno-Histoquímica , Lampreias/metabolismo , Células Fotorreceptoras/fisiologia , Animais , Lampreias/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Larva/metabolismo , Retina/crescimento & desenvolvimento , Retina/metabolismo
4.
FASEB J ; 35(9): e21846, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34405458

RESUMO

Myopia (short-sightedness), usually caused by excessive elongation of the eye during development, has reached epidemic proportions worldwide. In animal systems including the chicken model, several treatments have been shown to inhibit ocular elongation and experimental myopia. Although diverse in their apparent mechanism of action, each one leads to a reduction in the rate of ocular growth. We hypothesize that a defined set of retinal molecular changes may underlie growth inhibition, irrespective of the treatment agent used. Accordingly, across five well-established but diverse methods of inhibiting myopia, significant overlap is seen in the retinal transcriptome profile (transcript levels and alternative splicing events) in chicks when analyzed by RNA-seq. Within the two major pathway networks enriched during growth inhibition, that of cell signaling and circadian entrainment, transcription factors form the largest functional grouping. Importantly, a large percentage of those genes forming the defined retinal response are downstream targets of the transcription factor EGR1 which itself shows a universal response to all five growth-inhibitory treatments. This supports EGR1's previously implicated role in ocular growth regulation. Finally, by contrasting our data with human linkage and GWAS studies on refractive error, we confirm the applicability of our study to the human condition. Together, these findings suggest that a universal set of transcriptome changes, which sit within a well-defined retinal network that cannot be bypassed, is fundamental to growth regulation, thus paving a way for designing novel targets for myopia therapies.


Assuntos
Olho/crescimento & desenvolvimento , Olho/metabolismo , Redes Reguladoras de Genes , Miopia/genética , Miopia/prevenção & controle , Transcriptoma , Processamento Alternativo/efeitos dos fármacos , Animais , Atropina/farmacologia , Galinhas , Ritmo Circadiano/efeitos dos fármacos , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Olho/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Humanos , Janus Quinases/metabolismo , Masculino , Modelos Biológicos , Ácidos Fosfínicos/farmacologia , Pirenzepina/farmacologia , Piridinas/farmacologia , Reprodutibilidade dos Testes , Retina/efeitos dos fármacos , Retina/crescimento & desenvolvimento , Retina/metabolismo , Fatores de Transcrição STAT/metabolismo , Tetra-Hidronaftalenos/farmacologia , Fatores de Tempo , Transcriptoma/efeitos dos fármacos
5.
Elife ; 102021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34252023

RESUMO

Organoids derived from pluripotent stem cells promise the solution to current challenges in basic and biomedical research. Mammalian organoids are however limited by long developmental time, variable success, and lack of direct comparison to an in vivo reference. To overcome these limitations and address species-specific cellular organization, we derived organoids from rapidly developing teleosts. We demonstrate how primary embryonic pluripotent cells from medaka and zebrafish efficiently assemble into anterior neural structures, particularly retina. Within 4 days, blastula-stage cell aggregates reproducibly execute key steps of eye development: retinal specification, morphogenesis, and differentiation. The number of aggregated cells and genetic factors crucially impacted upon the concomitant morphological changes that were intriguingly reflecting the in vivo situation. High efficiency and rapid development of fish-derived organoids in combination with advanced genome editing techniques immediately allow addressing aspects of development and disease, and systematic probing of impact of the physical environment on morphogenesis and differentiation.


Assuntos
Células-Tronco Embrionárias/citologia , Organogênese , Organoides/citologia , Retina/citologia , Animais , Diferenciação Celular , Células-Tronco Embrionárias/metabolismo , Humanos , Morfogênese , Organoides/metabolismo , Oryzias , Células-Tronco Pluripotentes/fisiologia , Retina/crescimento & desenvolvimento , Retina/metabolismo , Peixe-Zebra
6.
Dev Biol ; 478: 144-154, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34260962

RESUMO

Throughout the central nervous system, astrocytes adopt precisely ordered spatial arrangements of their somata and arbors, which facilitate their many important functions. Astrocyte pattern formation is particularly important in the retina, where astrocytes serve as a template that dictates the pattern of developing retinal vasculature. Thus, if astrocyte patterning is disturbed, there are severe consequences for retinal angiogenesis and ultimately for vision - as seen in diseases such as retinopathy of prematurity. Here we discuss key steps in development of the retinal astrocyte population. We describe how fundamental developmental forces - their birth, migration, proliferation, and death - sculpt astrocytes into a template that guides angiogenesis. We further address the radical changes in the cellular and molecular composition of the astrocyte network that occur upon completion of angiogenesis, paving the way for their adult functions in support of retinal ganglion cell axons. Understanding development of retinal astrocytes may elucidate pattern formation mechanisms that are deployed broadly by other axon-associated astrocyte populations.


Assuntos
Astrócitos/fisiologia , Retina/crescimento & desenvolvimento , Retina/fisiologia , Animais , Axônios/fisiologia , Morte Celular , Diferenciação Celular , Movimento Celular , Proliferação de Células , Humanos , Neovascularização Fisiológica , Fibras Nervosas/fisiologia , Retina/citologia , Retina/embriologia , Células Ganglionares da Retina/fisiologia , Vasos Retinianos/embriologia , Vasos Retinianos/crescimento & desenvolvimento , Vasos Retinianos/fisiologia , Retinopatia da Prematuridade/patologia , Retinopatia da Prematuridade/fisiopatologia
7.
Invest Ophthalmol Vis Sci ; 62(9): 15, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34241625

RESUMO

Purpose: Primary cilia are conserved organelles found in polarized cells within the eye that regulate cell growth, migration, and differentiation. Although the role of cilia in photoreceptors is well-studied, the formation of cilia in other retinal cell types has received little attention. In this study, we examined the ciliary profile focused on the inner nuclear layer of retinas in mice and rhesus macaque primates. Methods: Retinal sections or flatmounts from Arl13b-Cetn2 tg transgenic mice were immunostained for cell markers (Pax6, Sox9, Chx10, Calbindin, Calretinin, ChaT, GAD67, Prox1, TH, and vGluT3) and analyzed by confocal microscopy. Primate retinal sections were immunostained for ciliary and cell markers (Pax6 and Arl13b). Optical coherence tomography (OCT) and ERGs were used to assess visual function of Vift88 mice. Results: During different stages of mouse postnatal eye development, we found that cilia are present in Pax6-positive amacrine cells, which were also observed in primate retinas. The cilia of subtypes of amacrine cells in mice were shown by immunostaining and electron microscopy. We also removed primary cilia from vGluT3 amacrine cells in mouse and found no significant vision defects. In addition, cilia were present in the outer limiting membrane, suggesting that a population of Müller glial cells forms cilia. Conclusions: We report that several subpopulations of amacrine cells in inner nuclear layers of the retina form cilia during early retinal development in mice and primates.


Assuntos
Células Amácrinas/ultraestrutura , Retina/crescimento & desenvolvimento , Animais , Galinhas , Cílios , Eletrorretinografia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Eletrônica , Modelos Animais , Coelhos , Retina/ultraestrutura , Tomografia de Coerência Óptica/métodos
8.
Int J Mol Sci ; 22(13)2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-34209226

RESUMO

As neurotransmitter, GABA is fundamental for physiological processes in the developing retina. Its synthesis enzymes are present during retinal development, although the molecular regulatory mechanisms behind the changes in expression are not entirely understood. In this study, we revealed the expression patterns of glutamic acid decarboxylase 67(GAD67) and its coding gene (GAD1) and its potential miRNA-dependent regulation during the first three postnatal weeks in rat retina. To gain insight into the molecular mechanisms, miRNA-sequencing supported by RT-qPCR and in situ hybridization were carried out. GAD1 expression shows an increasing tendency, peaking at P15. From the in silico-predicted GAD1 targeting miRNAs, only miR-23 showed similar expression patterns, which is a known regulator of GAD1 expression. For further investigation, we made an in situ hybridization investigation where both GAD67 and miR-23 also showed lower expression before P7, with the intensity of expression gradually increasing until P21. Horizontal cells at P7, amacrine cells at P15 and P21, and some cells in the ganglion cell layer at several time points were double labelled with miR-23 and GAD67. Our results highlight the complexity of these regulatory networks and the possible role of miR-23 in the regulation of GABA synthesizing enzyme expression during postnatal retina development.


Assuntos
Regulação Enzimológica da Expressão Gênica , Glutamato Descarboxilase/biossíntese , MicroRNAs/biossíntese , Retina/crescimento & desenvolvimento , Animais , Glutamato Descarboxilase/genética , MicroRNAs/genética , Ratos , Ratos Wistar
9.
Int J Mol Sci ; 22(14)2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34298993

RESUMO

Age-related macular degeneration (AMD) is a complex multifactorial neurodegenerative disease that constitutes the most common cause of irreversible blindness in the elderly in the developed countries. Incomplete knowledge about its pathogenesis prevents the search for effective methods of prevention and treatment of AMD, primarily of its "dry" type which is by far the most common (90% of all AMD cases). In the recent years, AMD has become "younger": late stages of the disease are now detected in relatively young people. It is known that AMD pathogenesis-according to the age-related structural and functional changes in the retina-is linked with inflammation, hypoxia, oxidative stress, mitochondrial dysfunction, and an impairment of neurotrophic support, but the mechanisms that trigger the conversion of normal age-related changes to the pathological process as well as the reason for early AMD development remain unclear. In the adult mammalian retina, de novo neurogenesis is very limited. Therefore, the structural and functional features that arise during its maturation and formation can exert long-term effects on further ontogenesis of this tissue. The aim of this review was to discuss possible contributions of the changes/disturbances in retinal neurogenesis to the early development of AMD.


Assuntos
Envelhecimento/patologia , Degeneração Macular/etiologia , Degeneração Macular/metabolismo , Doenças Neurodegenerativas/metabolismo , Neurogênese , Retina/crescimento & desenvolvimento , Envelhecimento/genética , Envelhecimento/metabolismo , Animais , Humanos , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Degeneração Macular/genética , Degeneração Macular/patologia , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Neurogênese/efeitos dos fármacos , Neurogênese/genética , Retina/patologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
10.
Invest Ophthalmol Vis Sci ; 62(7): 1, 2021 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-34061953

RESUMO

Purpose: The neuronal ELAV-like proteins (nElavls; Elavl2, Elavl3, Elavl4) have been known to regulate neuronal differentiation, maintenance, and axonogenesis in the brain. However, the specific role of nElavls in retina remains unclear. Here, we attempted to identify the expression pattern of Elavl2 during retinogenesis and aimed to decipher the function of Elavl2 in the retina. Methods: We have used the Cre-loxP system to conditionally inactivate Elavl2 in order to examine its role in developing retina. Eyes were collected for histology, immunohistochemistry, and TUNEL analysis to identify the structure of retina, and examined by RNA sequencing to analyze the function and pathway enrichment of differentially expressed genes in transgenic mice. Moreover, the mechanism by which Elavl2 regulates the differentiation of amacrine cells (ACs) was explored by RNA immunoprecipitation assays. Finally, eyes were functionally assessed by whole-cell patch-clamp, electroretinography (ERG) and optomotor response. Results: Elavl2 was expressed in retinal progenitor cells and retinal ganglion cells (RGCs), ACs, and horizontal cells. Retina-specific ablation of Elavl2 led to the loss of ACs and the transcription factors involved in ACs differentiation were also downregulated. In addition, the spontaneous activities of RGCs were obviously increased in Elavl2-deficient mice. Meanwhile, the loss of ACs that induced by Elavl2 deficiency lead to a decrease in ERG responses and visual acuity. Conclusions: Elavl2 is an intrinsic factor that involved in the differentiation of ACs subtype during retinogenesis, and essential for maintaining the normal retinal function.


Assuntos
Células Amácrinas/fisiologia , Proteína Semelhante a ELAV 2/genética , Proteínas do Tecido Nervoso/genética , Neurogênese/genética , Retina , Animais , Diferenciação Celular , Eletrorretinografia/métodos , Regulação da Expressão Gênica no Desenvolvimento , Interneurônios/fisiologia , Camundongos , Camundongos Transgênicos , Técnicas de Patch-Clamp/métodos , Retina/embriologia , Retina/crescimento & desenvolvimento , Retina/fisiologia , Células Ganglionares da Retina/fisiologia , Transcriptoma
11.
Dev Biol ; 478: 41-58, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34146533

RESUMO

Recent advances in high throughput single-cell RNA sequencing (scRNA-seq) technology have enabled the simultaneous transcriptomic profiling of thousands of individual cells in a single experiment. To investigate the intrinsic process of retinal development, researchers have leveraged this technology to quantify gene expression in retinal cells across development, in multiple species, and from numerous important models of human disease. In this review, we summarize recent applications of scRNA-seq and discuss how these datasets have complemented and advanced our understanding of retinal progenitor cell competence, cell fate specification, and differentiation. Finally, we also highlight the outstanding questions in the field that advances in single-cell data generation and analysis will soon be able to answer.


Assuntos
Células-Tronco Multipotentes/citologia , RNA-Seq , Retina/crescimento & desenvolvimento , Neurônios Retinianos/citologia , Análise de Célula Única , Animais , Linhagem da Célula , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Células-Tronco Multipotentes/metabolismo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Neurogênese , Retina/citologia , Retina/embriologia , Doenças Retinianas/genética , Doenças Retinianas/metabolismo , Doenças Retinianas/patologia , Neurônios Retinianos/metabolismo , Transcriptoma
12.
Dev Biol ; 478: 102-121, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34181916

RESUMO

Human organoids stand at the forefront of basic and translational research, providing experimentally tractable systems to study human development and disease. These stem cell-derived, in vitro cultures can generate a multitude of tissue and organ types, including distinct brain regions and sensory systems. Neural organoid systems have provided fundamental insights into molecular mechanisms governing cell fate specification and neural circuit assembly and serve as promising tools for drug discovery and understanding disease pathogenesis. In this review, we discuss several human neural organoid systems, how they are generated, advances in 3D imaging and bioengineering, and the impact of organoid studies on our understanding of the human nervous system.


Assuntos
Encefalopatias , Encéfalo , Organoides , Retina , Encéfalo/citologia , Encéfalo/embriologia , Encéfalo/crescimento & desenvolvimento , Diferenciação Celular , Biologia do Desenvolvimento/métodos , Corpos Embrioides/fisiologia , Indução Embrionária , Humanos , Células-Tronco Neurais/fisiologia , Neurobiologia/métodos , Neurogênese , Retina/citologia , Retina/embriologia , Retina/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
13.
Int J Mol Sci ; 22(9)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-34062733

RESUMO

Retinopathy of prematurity (ROP) is an ocular vascular disease affecting premature infants, characterized by pathological retinal neovascularization (RNV), dilated and tortuous retinal blood vessels, and retinal or vitreous hemorrhages that may lead to retinal detachment, vision impairment and blindness. Compared with other neovascular diseases, ROP is unique because of ongoing and concurrent physiological and pathological angiogenesis in the developing retina. While the disease is currently treated by laser or cryotherapy, anti-vascular endothelial growth factor (VEGF) agents have been extensively investigated but are not approved in the U.S. because of safety concerns that they negatively interfere with physiological angiogenesis of the developing retina. An ideal therapeutic strategy would selectively inhibit pathological but not physiological angiogenesis. Our group recently described a novel strategy that selectively and safely alleviates pathological RNV in animal models of ROP by targeting secretogranin III (Scg3), a disease-restricted angiogenic factor. The preclinical profile of anti-Scg3 therapy presents a high potential for next-generation disease-targeted anti-angiogenic therapy for the ROP indication. This review focuses on retinal vessel development in neonates, the pathogenesis of ROP and its underlying molecular mechanisms, including different animal models, and provides a summary of current and emerging therapies.


Assuntos
Cromograninas/genética , Neovascularização Patológica/tratamento farmacológico , Oxigênio/uso terapêutico , Retinopatia da Prematuridade/tratamento farmacológico , Animais , Animais Recém-Nascidos , Cromograninas/antagonistas & inibidores , Humanos , Camundongos , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/genética , Retina/efeitos dos fármacos , Retina/crescimento & desenvolvimento , Retina/patologia , Retinopatia da Prematuridade/genética , Retinopatia da Prematuridade/patologia , Fator A de Crescimento do Endotélio Vascular/genética
14.
Development ; 148(9)2021 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-33960384

RESUMO

Angiogenesis in the developing mammalian retina requires patterning cues from astrocytes. Developmental disorders of retinal vasculature, such as retinopathy of prematurity (ROP), involve arrest or mispatterning of angiogenesis. Whether these vascular pathologies involve astrocyte dysfunction remains untested. Here, we demonstrate that the major risk factor for ROP - transient neonatal exposure to excess oxygen - disrupts formation of the angiogenic astrocyte template. Exposing newborn mice to elevated oxygen (75%) suppressed astrocyte proliferation, whereas return to room air (21% oxygen) at postnatal day 4 triggered extensive proliferation, massively increasing astrocyte numbers and disturbing their spatial patterning prior to the arrival of developing vasculature. Proliferation required astrocytic HIF2α and was also stimulated by direct hypoxia (10% oxygen), suggesting that astrocyte oxygen sensing regulates the number of astrocytes produced during development. Along with astrocyte defects, return to room air also caused vascular defects reminiscent of ROP. Strikingly, these vascular phenotypes were more severe in animals that had larger numbers of excess astrocytes. Together, our findings suggest that fluctuations in environmental oxygen dysregulate molecular pathways controlling astrocyte proliferation, thereby generating excess astrocytes that interfere with retinal angiogenesis.


Assuntos
Astrócitos/metabolismo , Proliferação de Células/fisiologia , Neovascularização Patológica/metabolismo , Neovascularização Fisiológica/fisiologia , Oxigênio/metabolismo , Retina/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Hipóxia/metabolismo , Camundongos , Neovascularização Fisiológica/efeitos dos fármacos , Oxigênio/farmacologia , Retina/anormalidades , Retina/metabolismo , Retina/patologia , Vasos Retinianos/metabolismo , Retinopatia da Prematuridade
15.
Invest Ophthalmol Vis Sci ; 62(6): 1, 2021 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-33938913

RESUMO

Purpose: The trimethylation of histone H3 at lysine 4 (H3K4me3) facilitates transcriptional gene activation, and Setd1a is the methyltransferase specific to H3K4. H3K4me3 has been reported to regulate rod photoreceptor differentiation; however, the roles H3K4me3 plays in retinal progenitor cell (RPC) proliferation and differentiation during early retinal development remain unclear. Methods: Using an in vitro retinal explant culture system, we suppressed the expression of Setd1a by introducing shSetd1a. We examined the expression level and H3K4me3 level of genes by RNA Sequencing and ChIP assay, respectively. Results: We found that Setd1a depletion resulted in increased apoptosis and proliferation failure in late RPCs. Expression of wild-type SETD1A, but not SETD1A that lacked the catalytic SET domain, reversed the shSetd1a-induced phenotype. RNA Sequencing revealed that proliferation-related genes were downregulated upon shSetd1a expression. Based on publicly available H3K4me3-ChIP sequencing data of retinal development, we identified Uhrf1 as a candidate target gene of Setd1a. The expression of shSetd1a led to a decrease in Uhrf1 transcript levels and reduced H3K4me3 levels at the Uhrf1 locus. Increased apoptosis and the suppression of proliferation in late RPCs were observed in retinal explants expressing shUhrf1, similar to the outcomes observed in shSetd1a-expressing retinas. The overexpression of UHRF1 did not rescue shSetd1a-induced apoptosis, but reversed the suppression of proliferation. Conclusions: These results indicate that Setd1a contributes to the survival and proliferation of retinal cells by regulating histone methylation, Setd1a regulates Uhrf1 expression, and these two molecules cooperate to regulate RPC survival and proliferation.


Assuntos
Proteínas Estimuladoras de Ligação a CCAAT/genética , Proliferação de Células/fisiologia , Sobrevivência Celular/fisiologia , Regulação da Expressão Gênica/fisiologia , Histona-Lisina N-Metiltransferase/fisiologia , Retina/crescimento & desenvolvimento , Células-Tronco/citologia , Ubiquitina-Proteína Ligases/genética , Animais , Imunoprecipitação da Cromatina , Metilação de DNA/genética , Eletroporação , Histonas/genética , Imuno-Histoquímica , Camundongos Endogâmicos ICR , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA
16.
Int J Mol Sci ; 22(9)2021 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-33946979

RESUMO

Choline is essential for maintaining the structure and function of cells in humans. Choline plays an important role in eye health and disease. It is a precursor of acetylcholine, a neurotransmitter of the parasympathetic nervous system, and it is involved in the production and secretion of tears by the lacrimal glands. It also contributes to the stability of the cells and tears on the ocular surface and is involved in retinal development and differentiation. Choline deficiency is associated with retinal hemorrhage, glaucoma, and dry eye syndrome. Choline supplementation may be effective for treating these diseases.


Assuntos
Colina/fisiologia , Oftalmopatias/metabolismo , Acetilcolina/biossíntese , Acetilcolina/fisiologia , Animais , Deficiência de Colina/complicações , Deficiência de Colina/fisiopatologia , Retinopatia Diabética/fisiopatologia , Síndromes do Olho Seco/tratamento farmacológico , Síndromes do Olho Seco/metabolismo , Síndromes do Olho Seco/fisiopatologia , Oftalmopatias/etiologia , Oftalmopatias/fisiopatologia , Dor Ocular/fisiopatologia , Glaucoma/fisiopatologia , Glicerilfosforilcolina/uso terapêutico , Humanos , Aparelho Lacrimal/inervação , Aparelho Lacrimal/metabolismo , Cristalino/metabolismo , Nociceptividade/fisiologia , Nervo Óptico/metabolismo , Sistema Nervoso Parassimpático/fisiopatologia , Fosfatidilcolinas/biossíntese , Fosfolipídeos/metabolismo , Receptores Nicotínicos/fisiologia , Retina/crescimento & desenvolvimento , Retina/metabolismo , Vasos Retinianos/metabolismo , Lágrimas/metabolismo
17.
Invest Ophthalmol Vis Sci ; 62(4): 22, 2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33861321

RESUMO

Purpose: Light is a salient cue that can influence neurodevelopment and the immune system. Light exposure out of sync with the endogenous clock causes circadian disruption and chronic disease. Environmental light exposure may contribute to developmental programming of metabolic and neurological systems but has been largely overlooked in Developmental Origins of Health and Disease (DOHaD) research. Here, we investigated whether developmental light exposure altered programming of visual and metabolic systems. Methods: Pregnant mice and pups were exposed to control light (12:12 light:dark) or weekly light cycle inversions (circadian disruption [CD]) until weaning, after which male and female offspring were housed in control light and longitudinally measured to evaluate differences in growth (weight), glucose tolerance, visual function (optomotor response), and retinal function (electroretinogram), with and without high fat diet (HFD) challenge. Retinal microglia and macrophages were quantified by positive Iba1 and CD11b immunofluorescence. Results: CD exposure caused impaired visual function and increased retinal immune cell expression in adult offspring. When challenged with HFD, CD offspring also exhibited altered retinal function and sex-specific impairments in glucose tolerance. Conclusions: Overall, these findings suggest that the light environment contributes to developmental programming of the metabolic and visual systems, potentially promoting a pro-inflammatory milieu in the retina and increasing the risk of visual disease later in life.


Assuntos
Ritmo Circadiano/fisiologia , Luz , Prenhez , Retina/crescimento & desenvolvimento , Transtornos da Visão/fisiopatologia , Animais , Animais Recém-Nascidos , Feminino , Masculino , Camundongos Endogâmicos C57BL , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Retina/efeitos da radiação , Transtornos da Visão/etiologia
18.
Development ; 148(9)2021 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-33929509

RESUMO

The vertebrate retina is generated by retinal progenitor cells (RPCs), which produce >100 cell types. Although some RPCs produce many cell types, other RPCs produce restricted types of daughter cells, such as a cone photoreceptor and a horizontal cell (HC). We used genome-wide assays of chromatin structure to compare the profiles of a restricted cone/HC RPC and those of other RPCs in chicks. These data nominated regions of regulatory activity, which were tested in tissue, leading to the identification of many cis-regulatory modules (CRMs) active in cone/HC RPCs and developing cones. Two transcription factors, Otx2 and Oc1, were found to bind to many of these CRMs, including those near genes important for cone development and function, and their binding sites were required for activity. We also found that Otx2 has a predicted autoregulatory CRM. These results suggest that Otx2, Oc1 and possibly other Onecut proteins have a broad role in coordinating cone development and function. The many newly discovered CRMs for cones are potentially useful reagents for gene therapy of cone diseases.


Assuntos
Dissecação , Fator 6 Nuclear de Hepatócito/metabolismo , Fatores de Transcrição Otx/metabolismo , Retina/crescimento & desenvolvimento , Células Fotorreceptoras Retinianas Cones/metabolismo , Fatores de Transcrição/metabolismo , Animais , Sítios de Ligação , Galinhas , Cromatina , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Fator 6 Nuclear de Hepatócito/genética , Fatores de Transcrição Otx/genética , Retina/metabolismo , Células-Tronco
19.
PLoS Genet ; 17(4): e1009479, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33857132

RESUMO

Optogenetics has been harnessed to shed new mechanistic light on current and future therapeutic strategies. This has been to date achieved by the regulation of ion flow and electrical signals in neuronal cells and neural circuits that are known to be affected by disease. In contrast, the optogenetic delivery of trophic biochemical signals, which support cell survival and are implicated in degenerative disorders, has never been demonstrated in an animal model of disease. Here, we reengineered the human and Drosophila melanogaster REarranged during Transfection (hRET and dRET) receptors to be activated by light, creating one-component optogenetic tools termed Opto-hRET and Opto-dRET. Upon blue light stimulation, these receptors robustly induced the MAPK/ERK proliferative signaling pathway in cultured cells. In PINK1B9 flies that exhibit loss of PTEN-induced putative kinase 1 (PINK1), a kinase associated with familial Parkinson's disease (PD), light activation of Opto-dRET suppressed mitochondrial defects, tissue degeneration and behavioral deficits. In human cells with PINK1 loss-of-function, mitochondrial fragmentation was rescued using Opto-dRET via the PI3K/NF-кB pathway. Our results demonstrate that a light-activated receptor can ameliorate disease hallmarks in a genetic model of PD. The optogenetic delivery of trophic signals is cell type-specific and reversible and thus has the potential to inspire novel strategies towards a spatio-temporal regulation of tissue repair.


Assuntos
Proteínas de Drosophila/genética , Mitocôndrias/genética , Neurônios/metabolismo , Doença de Parkinson/genética , Proteínas Serina-Treonina Quinases/genética , Animais , Modelos Animais de Doenças , Drosophila melanogaster/genética , Humanos , Luz , Mutação com Perda de Função/genética , Mitocôndrias/efeitos da radiação , Neurônios/patologia , Neurônios/efeitos da radiação , Optogenética/métodos , Doença de Parkinson/patologia , Fosfatidilinositol 3-Quinases/genética , Retina/crescimento & desenvolvimento , Retina/metabolismo , Transdução de Sinais/genética , Transfecção
20.
Development ; 148(7)2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33722901

RESUMO

How the body and organs balance their relative growth is of key importance for coordinating size and function. This is of particular relevance in organisms, which continue to grow over their entire life span. We addressed this issue in the neuroretina of medaka fish (Oryzias latipes), a well-studied system with which to address vertebrate organ growth. We reveal that a central growth regulator, Igf1 receptor (Igf1r), is necessary and sufficient for proliferation control in the postembryonic retinal stem cell niche: the ciliary marginal zone (CMZ). Targeted activation of Igf1r signaling in the CMZ uncouples neuroretina growth from body size control, and we demonstrate that Igf1r operates on progenitor cells, stimulating their proliferation. Activation of Igf1r signaling increases retinal size while preserving its structural integrity, revealing a modular organization in which progenitor differentiation and neurogenesis are self-organized and highly regulated. Our findings position Igf signaling as a key module for controlling retinal size and composition, with important evolutionary implications.


Assuntos
Fator de Crescimento Insulin-Like I/metabolismo , Oryzias/crescimento & desenvolvimento , Receptor IGF Tipo 1/metabolismo , Retina/crescimento & desenvolvimento , Transdução de Sinais , Células-Tronco/fisiologia , Animais , Animais Geneticamente Modificados , Ciclo Celular , Diferenciação Celular/fisiologia , Divisão Celular/fisiologia , Proliferação de Células , Autorrenovação Celular , Fator de Crescimento Insulin-Like I/genética , Neurogênese , Oryzias/embriologia , Oryzias/genética , Receptor IGF Tipo 1/genética , Retina/citologia , Nicho de Células-Tronco , Células-Tronco/citologia , Vertebrados
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