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1.
J Cell Sci ; 124(Pt 7): 1017-21, 2011 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-21402875

RESUMEN

Collective cell migration is absolutely essential for a wide variety of physiological episodes including the re-epithelialisation component of tissue repair. However, the investigation of such processes has been frustrated by difficulties in quantitatively analysing the behaviours of a large body of cells within a migrating epithelial sheet, which previously required manually tracking a large number of individual cells, or using advanced computational techniques. Here, we describe a novel and simpler image subtraction method with which we can visualise and quantify collective cell mobilisation as a 'white wave' that propagates back from the leading edge of a scratch-wounded monolayer of cultured epithelial cells. Using this technique, we show that actomyosin constriction negatively regulates cell mobilisation and that the advancement of cell sheets and the mobilisation of rows of cells behind their leading edges are independently regulated. We also show that there is a finite limit to the number of rows of cells mobilised after wounding. Moreover, our data suggest that enhancing cell mobilisation, by release from myosin II contractility, accelerates the healing of large wounds in the long term, thus raising the possibility that the cell mobilisation 'wave' we reveal here might be a therapeutic target for improving wound healing.


Asunto(s)
Movimiento Celular , Células Epiteliales/citología , Células Epiteliales/metabolismo , Miosina Tipo II/metabolismo , Cicatrización de Heridas , Animales , Células Cultivadas , Ratones , Miosina Tipo II/genética
2.
Fly (Austin) ; 16(1): 248-274, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-35856387

RESUMEN

Extracellular matrices (ECMs) are essential for the architecture and function of animal tissues. ECMs have been thought to be highly stable structures; however, too much stability of ECMs would hamper tissue remodelling required for organ development and maintenance. Regarding this conundrum, this article reviews multiple lines of evidence that ECMs are in fact rapidly moving and replacing components in diverse organisms including hydra, worms, flies, and vertebrates. Also discussed are how cells behave on/in such dynamic ECMs, how ECM dynamics contributes to embryogenesis and adult tissue homoeostasis, and what molecular mechanisms exist behind the dynamics. In addition, it is highlighted how cutting-edge technologies such as genome engineering, live imaging, and mathematical modelling have contributed to reveal the previously invisible dynamics of ECMs. The idea that ECMs are unchanging is to be changed, and ECM dynamics is emerging as a hitherto unrecognized critical factor for tissue development and maintenance.


Asunto(s)
Matriz Extracelular , Animales , Matriz Extracelular/química
3.
J Neurosci ; 30(8): 3082-92, 2010 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-20181605

RESUMEN

Elucidating neuronal circuits and their plasticity in the cerebral cortex is one of the important questions in neuroscience research. Here we report novel axonal trajectories and their plasticity in the mouse somatosensory barrel cortex. We selectively visualized layer 2/3 neurons using in utero electroporation and examined the axonal trajectories of layer 2/3 neurons. We found that the axons of layer 2/3 neurons preferentially run in the septal regions of layer 4 and named this axonal pattern "barrel nets." The intensity of green fluorescent protein in the septal regions was markedly higher compared with that in barrel hollows. Focal in utero electroporation revealed that the axons in barrel nets were indeed derived from layer 2/3 neurons in the barrel cortex. During development, barrel nets became visible at postnatal day 10, which was well after the initial appearance of barrels. When whisker follicles were cauterized within 3 d after birth, the whisker-related pattern of barrel nets was altered, suggesting that cauterization of whisker follicles results in developmental plasticity of barrel nets. Our results uncover the novel axonal trajectories of layer 2/3 neurons with whisker-related patterns and their developmental plasticity in the mouse somatosensory cortex. Barrel nets should be useful for investigating the pattern formation and axonal reorganization of intracortical neuronal circuits.


Asunto(s)
Axones/fisiología , Plasticidad Neuronal/fisiología , Células Receptoras Sensoriales/fisiología , Corteza Somatosensorial/crecimiento & desarrollo , Nervio Trigémino/fisiología , Vibrisas/fisiología , Vías Aferentes/citología , Vías Aferentes/crecimiento & desarrollo , Animales , Axones/ultraestructura , Biomarcadores , Mapeo Encefálico , Electroporación , Proteínas Fluorescentes Verdes , Mecanorreceptores/fisiología , Mecanotransducción Celular/fisiología , Ratones , Ratones Endogámicos ICR , Microscopía Confocal , Neurópilo/citología , Neurópilo/fisiología , Proteínas Recombinantes de Fusión , Células Receptoras Sensoriales/citología , Corteza Somatosensorial/citología , Coloración y Etiquetado , Sinapsis/fisiología , Sinapsis/ultraestructura , Sinaptofisina , Percepción del Tacto/fisiología
4.
Dev Cell ; 56(15): 2137-2139, 2021 08 09.
Artículo en Inglés | MEDLINE | ID: mdl-34375578

RESUMEN

Epithelia have an innate yet mysterious capacity to rapidly sense and respond to tissue damage. In this issue of Developmental Cell, O'Connor et al. exploit the genetics of Drosophila to reveal that protease release as a result of tissue injury activates insect cytokines to initiate immediate epithelial repair responses.


Asunto(s)
Drosophila , Péptido Hidrolasas , Animales , Epitelio
5.
STAR Protoc ; 2(1): 100377, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33786460

RESUMEN

Protein turnover rate is difficult to obtain experimentally. This protocol shows how to mathematically model turnover rates in an intervention-free manner given the ability to quantify mRNA and protein expression from initiation to homeostasis. This approach can be used to calculate production and degradation rates and to infer protein half-life. This model was successfully employed to quantify turnover during Drosophila embryogenesis, and we hypothesize that it will be applicable to diverse in vivo or in vitro systems. For complete details on the use and execution of this protocol, please refer to Matsubayashi et al. (2020).


Asunto(s)
Biología Computacional/métodos , Proteolisis , ARN Mensajero/metabolismo , Animales , Drosophila/metabolismo , Expresión Génica/genética , Homeostasis , Cinética , Modelos Teóricos , Proteínas/metabolismo
6.
Dev Cell ; 54(1): 33-42.e9, 2020 07 06.
Artículo en Inglés | MEDLINE | ID: mdl-32585131

RESUMEN

The extracellular matrix (ECM) is a polymer network hypothesized to form a stable cellular scaffold. While the ECM can undergo acute remodeling during embryogenesis, it is experimentally difficult to determine whether basal turnover is also important. Most studies of homeostatic turnover assume an initial steady-state balance of production and degradation and measure half-life by quantifying the rate of decay after experimental intervention (e.g., pulse labeling). Here, we present an intervention-free approach to mathematically model basal ECM turnover during embryogenesis by exploiting our ability to live image de novo ECM development in Drosophila to quantify production from initiation to homeostasis. This reveals rapid turnover (half-life ∼7-10 h), which we confirmed by in vivo pulse-chase experiments. Moreover, ECM turnover is partially dependent on proteolysis and network interactions, and slowing turnover affects tissue morphogenesis. These data demonstrate that embryonic ECM undergoes constant replacement, which is likely necessary to maintain network plasticity to accommodate growth and morphogenesis.


Asunto(s)
Matriz Extracelular/metabolismo , Homeostasis , Morfogénesis , Animales , Membrana Basal/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Células Epiteliales/citología , Células Epiteliales/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Modelos Teóricos
7.
J Neurosci Methods ; 178(1): 87-98, 2009 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-19100769

RESUMEN

Galactocerebroside (GalC) and its sulfated derivative sulfatide (SUL) are galactosphingolipids abundantly expressed in oligodendrocytes (OLs). Despite their biological importance in OL development and function, attempts to visualize GalC/SUL in tissue sections have met with limited success. This is at least in part because permeabilization of tissue sections with detergents such as Triton X-100 results in significant degradation of GalC/SUL immunoreactivity. Here we establish a novel method that enables visualization of endogenous GalC/SUL in OLs and myelin throughout the entire depth of brain sections. We show that treating brain sections with the cholesterol-specific detergent digitonin instead of Triton X-100 or methanol leads to efficient antibody penetration into tissue sections without disrupting GalC/SUL immunoreactivity. We also determine the optimal concentrations of digitonin using confocal microscopy. With our method, the morphology and the number of GalC/SUL-expressing OLs can be visualized three-dimensionally. Furthermore, our method is applicable to double immunostaining with anti-GalC/SUL antibody and other antibodies which recognize intracellular antigens. Our simple method using digitonin should prove to be useful in enabling detailed examination of GalC/SUL expression in the brain in both physiological and pathological conditions.


Asunto(s)
Encéfalo/citología , Detergentes/farmacología , Digitonina/farmacología , Galactosilceramidas/metabolismo , Oligodendroglía/efectos de los fármacos , Oligodendroglía/metabolismo , Animales , Animales Recién Nacidos , Proteínas Relacionadas con la Autofagia , Relación Dosis-Respuesta a Droga , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Proteína Básica de Mielina/metabolismo , Antígenos O/metabolismo , Octoxinol/farmacología , Fosfopiruvato Hidratasa/metabolismo , Sulfoglicoesfingolípidos/metabolismo
8.
Mol Cell Neurosci ; 39(1): 40-9, 2008 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-18588982

RESUMEN

Termination of developmental plasticity occurs at specific points in development, and the mechanisms responsible for it are not well understood. One hypothesis that has been proposed is that oligodendrocytes (OLs) play an important role. Consistent with this, we found that OLs appeared in the mouse somatosensory cortex at the end of the critical period for whisker lesion-induced barrel structural plasticity. To test this hypothesis, we used two mouse lines with defective OL differentiation: Olig1-deficient and jimpy. In Olig1-deficient mice, although OLs were totally absent, the termination of lesion-induced plasticity was not delayed. The timing was normal even when the cytoarchitectonic barrel formation was temporarily blocked by pharmacological treatment in Olig1-deficient mice. Furthermore, the termination was not delayed in jimpy mice. These results demonstrate that, even though OLs appear at the end of the critical period, OLs are not intrinsically necessary for the termination of lesion-induced plasticity. Our findings underscore a mechanistic distinction between the termination of thalamocortical axonal plasticity in the barrel cortex and that in the visual cortex, in which OL-derived Nogo-A/B was recently suggested to be essential.


Asunto(s)
Plasticidad Neuronal/fisiología , Oligodendroglía/metabolismo , Corteza Somatosensorial , Vibrisas/patología , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diferenciación Celular/fisiología , Clorgilina/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Jimpy , Ratones Noqueados , Inhibidores de la Monoaminooxidasa/metabolismo , Oligodendroglía/citología , Corteza Somatosensorial/citología , Corteza Somatosensorial/patología , Corteza Somatosensorial/fisiología
9.
J Neurosci Methods ; 174(1): 71-81, 2008 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-18674563

RESUMEN

The fluorescent carbocyanine dye DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) has been widely used for tracing of neuronal pathways. To examine identities of the DiI-labeled neuronal pathways, it is desirable to combine DiI labeling with immunofluorescent staining. However, DiI labeling and immunofluorescent staining are not well compatible, mainly because treatment of DiI-labeled neurons with detergents, which are commonly used for immunohistochemistry, results in high levels of diffusion of the DiI label. In this study, we searched for detergents that are compatible with DiI labeling, and found that a cholesterol-specific detergent digitonin is useful for fluorescent double-labeling with DiI tracing and immunohistochemistry. We show that digitonin treatment, in contrast to Triton X-100, methanol and Nonidet P-40 treatment, preserves DiI labeling, even at higher concentrations. We also show that digitonin also preserves the signal of a DiI derivative CM-DiI. Moreover, we demonstrate that digitonin efficiently increases antibody penetration into brain sections. As a result, immunohistochemical images obtained with digitonin treatment are as good as those obtained with Triton X-100 treatment. In addition, we also try another cholesterol-specific detergent quillaja saponin, but find that it degrades the DiI label. Our simple double-labeling protocol using digitonin should prove useful in enabling detailed examination of the neuronal circuitry of the nervous system.


Asunto(s)
Carbocianinas/química , Digitonina/química , Colorantes Fluorescentes/química , Inmunohistoquímica/métodos , Neuronas/citología , Coloración y Etiquetado/métodos , Animales , Anticuerpos/química , Anticuerpos/farmacología , Mapeo Encefálico/instrumentación , Mapeo Encefálico/métodos , Membrana Celular/química , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Colesterol/química , Colesterol/metabolismo , Detergentes/química , Difusión , Indicadores y Reactivos/química , Ratones , Ratones Endogámicos ICR , Microscopía Fluorescente/métodos , Neuroanatomía/instrumentación , Neuroanatomía/métodos , Neuronas/metabolismo , Fijación del Tejido/métodos
10.
G3 (Bethesda) ; 8(3): 845-857, 2018 03 02.
Artículo en Inglés | MEDLINE | ID: mdl-29321168

RESUMEN

Drosophila melanogaster plasmatocytes, the phagocytic cells among hemocytes, are essential for immune responses, but also play key roles from early development to death through their interactions with other cell types. They regulate homeostasis and signaling during development, stem cell proliferation, metabolism, cancer, wound responses, and aging, displaying intriguing molecular and functional conservation with vertebrate macrophages. Given the relative ease of genetics in Drosophila compared to vertebrates, tools permitting visualization and genetic manipulation of plasmatocytes and surrounding tissues independently at all stages would greatly aid a fuller understanding of these processes, but are lacking. Here, we describe a comprehensive set of transgenic lines that allow this. These include extremely brightly fluorescing mCherry-based lines that allow GAL4-independent visualization of plasmatocyte nuclei, the cytoplasm, or the actin cytoskeleton from embryonic stage 8 through adulthood in both live and fixed samples even as heterozygotes, greatly facilitating screening. These lines allow live visualization and tracking of embryonic plasmatocytes, as well as larval plasmatocytes residing at the body wall or flowing with the surrounding hemolymph. With confocal imaging, interactions of plasmatocytes and inner tissues can be seen in live or fixed embryos, larvae, and adults. They permit efficient GAL4-independent Fluorescence-Activated Cell Sorting (FACS) analysis/sorting of plasmatocytes throughout life. To facilitate genetic studies of reciprocal signaling, we have also made a plasmatocyte-expressing QF2 line that, in combination with extant GAL4 drivers, allows independent genetic manipulation of both plasmatocytes and surrounding tissues, and GAL80 lines that block GAL4 drivers from affecting plasmatocytes, all of which function from the early embryo to the adult.

11.
EMBO Mol Med ; 10(10)2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-30171089

RESUMEN

Argonaute 2 bound mature microRNA (Ago2-miRNA) complexes are key regulators of the wound inflammatory response and function in the translational processing of target mRNAs. In this study, we identified four wound inflammation-related Ago2-miRNAs (miR-139-5p, miR-142-3p, miR-142-5p, and miR-223) and show that miR-223 is critical for infection control. miR-223Y/- mice exhibited delayed sterile healing with prolonged neutrophil activation and interleukin-6 expression, and markedly improved repair of Staphylococcus aureus-infected wounds. We also showed that the expression of miR-223 was regulated by CCAAT/enhancer binding protein alpha in human neutrophils after exposure to S. aureus peptides. Treatment with miR-223Y/--derived neutrophils, or miR-223 antisense oligodeoxynucleotides in S. aureus-infected wild-type wounds markedly improved the healing of these otherwise chronic, slow healing wounds. This study reveals how miR-223 regulates the bactericidal capacity of neutrophils at wound sites and indicates that targeting miR-223 might be of therapeutic benefit for infected wounds in the clinic.


Asunto(s)
Inflamación/fisiopatología , MicroARNs/metabolismo , Neutrófilos/inmunología , Infecciones Estafilocócicas/fisiopatología , Staphylococcus aureus/inmunología , Infección de Heridas/fisiopatología , Animales , Células Cultivadas , Humanos , Ratones , Ratones Noqueados , MicroARNs/genética
12.
Curr Biol ; 14(8): 731-5, 2004 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-15084290

RESUMEN

In epithelial cell movements, which occur during wound healing or embryonic morphogenesis, sheets of cells move together as a unit. Molecular mechanisms that regulate this sheet movement have been largely unknown, although cell locomotion or movement mechanisms for individual cells, such as for fibroblastic cells, have been extensively studied. Here, we show that, during wound healing, sheets of MDCK epithelial cells migrate coordinately as a unit, and wound-induced activation of ERK MAP kinase (ERK1/2) propagates in cell sheets in accordance with the cell sheet movement. Inhibition of ERK1/2 activation by specific MEK inhibitors or by expressing dominant-negative ERK2 results in marked inhibition of the sheet movement during wound healing, and inhibition of the cell sheet movement by disrupting actin cytoskeleton suppresses propagation of ERK1/2 activation. These results indicate that cell movement and ERK1/2 activation form a positive feedback loop, which facilitates cell sheet migration. Moreover, we find that Src family kinase inhibitors suppress both cell migration and propagation of ERK1/2 activation, suggesting that Src family kinase may participate in this feedback loop. Interestingly, neither cell sheet migration as a unit nor migration-dependent propagation of ERK1/2 activation occurs during wound healing in fibroblastic 3Y1 cells. Thus, our results identify specific requirements of ERK1/2 MAP kinase for epithelial cell sheet movement.


Asunto(s)
Movimiento Celular/fisiología , Citoesqueleto/fisiología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/fisiología , Cicatrización de Heridas/fisiología , Animales , Anticuerpos Monoclonales , Butadienos/metabolismo , Células Cultivadas , Perros , Activación Enzimática , Epitelio/metabolismo , Retroalimentación Fisiológica , Fibroblastos , Flavonoides/metabolismo , Proteínas Fluorescentes Verdes , Proteínas Luminiscentes/metabolismo , Nitrilos/metabolismo , Ratas , Transfección , Familia-src Quinasas/metabolismo
13.
Curr Biol ; 27(22): 3526-3534.e4, 2017 Nov 20.
Artículo en Inglés | MEDLINE | ID: mdl-29129537

RESUMEN

The basement membrane (BM) is a thin layer of extracellular matrix (ECM) beneath nearly all epithelial cell types that is critical for cellular and tissue function. It is composed of numerous components conserved among all bilaterians [1]; however, it is unknown how all of these components are generated and subsequently constructed to form a fully mature BM in the living animal. Although BM formation is thought to simply involve a process of self-assembly [2], this concept suffers from a number of logistical issues when considering its construction in vivo. First, incorporation of BM components appears to be hierarchical [3-5], yet it is unclear whether their production during embryogenesis must also be regulated in a temporal fashion. Second, many BM proteins are produced not only by the cells residing on the BM but also by surrounding cell types [6-9], and it is unclear how large, possibly insoluble protein complexes [10] are delivered into the matrix. Here we exploit our ability to live image and genetically dissect de novo BM formation during Drosophila development. This reveals that there is a temporal hierarchy of BM protein production that is essential for proper component incorporation. Furthermore, we show that BM components require secretion by migrating macrophages (hemocytes) during their developmental dispersal, which is critical for embryogenesis. Indeed, hemocyte migration is essential to deliver a subset of ECM components evenly throughout the embryo. This reveals that de novo BM construction requires a combination of both production and distribution logistics allowing for the timely delivery of core components.


Asunto(s)
Membrana Basal/fisiología , Matriz Extracelular/metabolismo , Animales , Membrana Basal/metabolismo , Movimiento Celular/fisiología , Colágeno/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Células Epiteliales/metabolismo , Matriz Extracelular/fisiología , Macrófagos/metabolismo
14.
Adv Wound Care (New Rochelle) ; 5(6): 243-250, 2016 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-27274434

RESUMEN

Significance: The epidermis provides the main barrier function of skin, and therefore its repair following wounding is an essential component of wound healing. Repair of the epidermis, also known as reepithelialization, occurs by collective migration of epithelial cells from around the wound edge across the wound until the advancing edges meet and fuse. Therapeutic manipulation of this process could potentially be used to accelerate wound healing. Recent Advances: It is difficult to analyze the cellular and molecular mechanisms of reepithelialization in human tissue, so a variety of model organisms have been used to improve our understanding of the process. One model system that has been especially useful is the embryo of the fruit fly Drosophila, which provides a simple, accessible model of the epidermis and can be manipulated genetically, allowing detailed analysis of reepithelialization at the molecular level. This review will highlight the key insights that have been gained from studying reepithelialization in Drosophila embryos. Critical Issues: Slow reepithelialization increases the risk of wounds becoming infected and ulcerous; therefore, the development of therapies to accelerate or enhance the process would be a great clinical advance. Improving our understanding of the molecular mechanisms that underlie reepithelialization will help in the development of such therapies. Future Directions: Research in Drosophila embryos has identified a variety of genes and proteins involved in triggering and driving reepithelialization, many of which are conserved in humans. These novel reepithelialization proteins are potential therapeutic targets and therefore findings obtained in Drosophila may ultimately lead to significant clinical advances.

15.
J Cell Biol ; 210(3): 419-33, 2015 Aug 03.
Artículo en Inglés | MEDLINE | ID: mdl-26216900

RESUMEN

The ability to heal wounds efficiently is essential for life. After wounding of an epithelium, the cells bordering the wound form dynamic actin protrusions and/or a contractile actomyosin cable, and these actin structures drive wound closure. Despite their importance in wound healing, the molecular mechanisms that regulate the assembly of these actin structures at wound edges are not well understood. In this paper, using Drosophila melanogaster embryos, we demonstrate that Diaphanous, SCAR, and WASp play distinct but overlapping roles in regulating actin assembly during wound healing. Moreover, we show that endocytosis is essential for wound edge actin assembly and wound closure. We identify adherens junctions (AJs) as a key target of endocytosis during wound healing and propose that endocytic remodeling of AJs is required to form "signaling centers" along the wound edge that control actin assembly. We conclude that coordination of actin assembly, AJ remodeling, and membrane traffic is required for the construction of a motile leading edge during wound healing.


Asunto(s)
Actinas/metabolismo , Uniones Adherentes/metabolismo , Endocitosis/fisiología , Cicatrización de Heridas/fisiología , Animales , Cadherinas/genética , Proteínas Portadoras/metabolismo , Clatrina/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Epitelio/metabolismo , Forminas , Proteínas de Microfilamentos/metabolismo , Transporte de Proteínas/fisiología , Transducción de Señal/fisiología , Proteína del Síndrome de Wiskott-Aldrich/metabolismo
18.
Genes Cells ; 9(2): 83-93, 2004 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-15009089

RESUMEN

Microtubules are involved in many cellular events during the cell cycle and also in a variety of early embryonic developmental processes. Their architecture and properties change dramatically during the cell cycle and are properly regulated. However, these regulatory mechanisms have not been fully elucidated. C05D11.3 gene of Caenorhabditis elegans encodes a low molecular weight protein that is evolutionarily conserved from yeasts to mammals. A mouse homolog of the C05D11.3 product, APACD (ATP binding protein associated with cell differentiation), contains a thioredoxin-like domain and P-loop, and is present in both the nucleus and the cytoplasm, showing often localization to centrosomes and midbody. In C. elegans, C05D11.3 is expressed throughout development with higher levels of expression in most cells of the nervous system and in vulva. C05D11.3 RNAi-treated embryos show apparent defects in pronuclear migration or nuclear-centrosome rotation, and exhibit little astral microtubules and defective small spindles. These results indicate that C05D11.3, an evolutionarily conserved gene, is essential for proper microtubule organization and function in C. elegans. This gene family may be a conserved regulator of microtubule dynamics and function.


Asunto(s)
Proteínas de Caenorhabditis elegans/fisiología , Caenorhabditis elegans/embriología , Evolución Molecular , Genes de Helminto/genética , Microtúbulos/ultraestructura , Tubulina (Proteína)/fisiología , Secuencia de Aminoácidos , Animales , Caenorhabditis elegans/genética , Caenorhabditis elegans/ultraestructura , Proteínas de Caenorhabditis elegans/genética , División Celular/efectos de los fármacos , División Celular/genética , Núcleo Celular/química , Núcleo Celular/ultraestructura , Centrosoma/ultraestructura , Secuencia Conservada/genética , Técnica del Anticuerpo Fluorescente , Expresión Génica , Proteínas Fluorescentes Verdes , Células HeLa , Humanos , Estadios del Ciclo de Vida/genética , Proteínas Luminiscentes/análisis , Proteínas Luminiscentes/genética , Ratones , Microtúbulos/efectos de los fármacos , Microtúbulos/metabolismo , Datos de Secuencia Molecular , Proteínas Nucleares/genética , ARN Interferente Pequeño/farmacología , Ratas , Alineación de Secuencia , Tubulina (Proteína)/genética
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