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1.
J Exp Biol ; 227(9)2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38634259

RESUMO

Ex vivo physiological experiments using small insect models such as Drosophila larvae have become increasingly useful to address fundamental biological questions. To perform such experiments, various artificial saline solutions have been developed, but their osmolality varies significantly from one to the next. Such a variation of osmolality stems, in part, from the difficulty of determining the true value of haemolymph osmolality in Drosophila larvae. Thus, there is a pressing need to refine protocols for collecting and measuring the osmolality of the larval haemolymph. Two major obstacles are thought to impede the accurate analysis of haemolymph collected from small insects: melanin formation and gut-derived contamination. Here, we greatly refined existing haemolymph collection methods, evaluated the purity of the collected haemolymph under melanin-free conditions, and concluded that the true value of haemolymph osmolality is close to 306.0 mOsm kg-1 in Drosophila larvae.


Assuntos
Hemolinfa , Larva , Animais , Hemolinfa/química , Hemolinfa/metabolismo , Concentração Osmolar , Larva/crescimento & desenvolvimento , Larva/química , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Melaninas/metabolismo , Melaninas/análise
2.
Elife ; 122023 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-37309249

RESUMO

Appropriate modulation of escape behaviors in response to potentially damaging stimuli is essential for survival. Although nociceptive circuitry has been studied, it is poorly understood how genetic contexts affect relevant escape responses. Using an unbiased genome-wide association analysis, we identified an Ly6/α-neurotoxin family protein, Belly roll (Bero), which negatively regulates Drosophila nociceptive escape behavior. We show that Bero is expressed in abdominal leucokinin-producing neurons (ABLK neurons) and bero knockdown in ABLK neurons resulted in enhanced escape behavior. Furthermore, we demonstrated that ABLK neurons responded to activation of nociceptors and initiated the behavior. Notably, bero knockdown reduced persistent neuronal activity and increased evoked nociceptive responses in ABLK neurons. Our findings reveal that Bero modulates an escape response by regulating distinct neuronal activities in ABLK neurons.


Assuntos
Drosophila melanogaster , Estudo de Associação Genômica Ampla , Animais , Nociceptividade , Interneurônios , Neurônios , Drosophila , Neurotoxinas
3.
Elife ; 122023 01 17.
Artigo em Inglês | MEDLINE | ID: mdl-36647607

RESUMO

Nutrition in early life has profound effects on an organism, altering processes such as organogenesis. However, little is known about how specific nutrients affect neuronal development. Dendrites of class IV dendritic arborization neurons in Drosophila larvae become more complex when the larvae are reared on a low-yeast diet compared to a high-yeast diet. Our systematic search for key nutrients revealed that the neurons increase their dendritic terminal densities in response to a combined deficiency in vitamins, metal ions, and cholesterol. The deficiency of these nutrients upregulates Wingless in a closely located tissue, body wall muscle. Muscle-derived Wingless activates Akt in the neurons through the receptor tyrosine kinase Ror, which promotes the dendrite branching. In larval muscles, the expression of wingless is regulated not only in this key nutrient-dependent manner, but also by the JAK/STAT signaling pathway. Additionally, the low-yeast diet blunts neuronal light responsiveness and light avoidance behavior, which may help larvae optimize their survival strategies under low-nutritional conditions. Together, our studies illustrate how the availability of specific nutrients affects neuronal development through inter-organ signaling.


Assuntos
Dendritos , Proteínas de Drosophila , Animais , Dendritos/fisiologia , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Neurônios/fisiologia , Nutrientes , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais
4.
Genes Cells ; 28(3): 175-187, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36562594

RESUMO

In vivo, cells collectively migrate in a variety of developmental and pathological contexts. Coordinated epithelial rotation represents a unique type of collective cell migrations, which has been modeled in vitro under spatially confined conditions. Although it is known that the coordinated rotation depends on intercellular interactions, the contribution of E-cadherin, a major cell-cell adhesion molecule, has not been directly addressed on two-dimensional (2D) confined substrates. Here, using well-controlled fibronectin-coated surfaces, we tracked and compared the migratory behaviors of MDCK cells expressing or lacking E-cadherin. We observed that wild-type MDCK II cells exhibited persistent and coordinated rotations on discoidal patterns, while E-cadherin knockout cells migrated in a less coordinated manner without large-scale rotation. Our comparison of the collective dynamics between these two cell types revealed a series of changes in migratory behavior caused by the loss of E-cadherin, including a decreased global migration speed, less regularity in quantified coordination, and increased average density of topological defects. Taken together, these data demonstrate that spontaneous initiation of collective epithelial rotations depends on E-cadherin under 2D discoidal confinements.


Assuntos
Caderinas , Células Epiteliais , Animais , Cães , Caderinas/metabolismo , Adesão Celular , Células Madin Darby de Rim Canino , Movimento Celular , Células Epiteliais/metabolismo
5.
Elife ; 62017 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-29035200

RESUMO

In Drosophila larvae, Class IV sensory neurons respond to noxious thermal stimuli and provoke heat avoidance behavior. Previously, we showed that the activated neurons displayed characteristic fluctuations of firing rates, which consisted of repetitive high-frequency spike trains and subsequent pause periods, and we proposed that the firing rate fluctuations enhanced the heat avoidance (Terada et al., 2016). Here, we further substantiate this idea by showing that the pause periods and the frequency of fluctuations are regulated by small conductance Ca2+-activated K+ (SK) channels, and the SK knockdown larvae display faster heat avoidance than control larvae. The regulatory mechanism of the fluctuations in the Class IV neurons resembles that in mammalian Purkinje cells, which display complex spikes. Furthermore, our results suggest that such fluctuation coding in Class IV neurons is required to convert noxious thermal inputs into effective stereotyped behavior as well as general rate coding.


Assuntos
Potenciais de Ação , Drosophila , Nociceptores/fisiologia , Canais de Potássio Ativados por Cálcio de Condutância Baixa/metabolismo , Animais , Técnicas de Silenciamento de Genes , Larva , Canais de Potássio Ativados por Cálcio de Condutância Baixa/genética , Resposta Táctica
6.
J Cell Biol ; 216(3): 815-834, 2017 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-28209644

RESUMO

Mitochondria are key contributors to the etiology of diseases associated with neuromuscular defects or neurodegeneration. How changes in cellular metabolism specifically impact neuronal intracellular processes and cause neuropathological events is still unclear. We here dissect the molecular mechanism by which mitochondrial dysfunction induced by Prel aberrant function mediates selective dendritic loss in Drosophila melanogaster class IV dendritic arborization neurons. Using in vivo ATP imaging, we found that neuronal cellular ATP levels during development are not correlated with the progression of dendritic loss. We searched for mitochondrial stress signaling pathways that induce dendritic loss and found that mitochondrial dysfunction is associated with increased eIF2α phosphorylation, which is sufficient to induce dendritic pathology in class IV arborization neurons. We also observed that eIF2α phosphorylation mediates dendritic loss when mitochondrial dysfunction results from other genetic perturbations. Furthermore, mitochondrial dysfunction induces translation repression in class IV neurons in an eIF2α phosphorylation-dependent manner, suggesting that differential translation attenuation among neuron subtypes is a determinant of preferential vulnerability.


Assuntos
Fator de Iniciação 2 em Eucariotos/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Doenças Mitocondriais/metabolismo , Doenças Mitocondriais/patologia , Fosforilação/fisiologia , Trifosfato de Adenosina/metabolismo , Animais , Dendritos/metabolismo , Dendritos/patologia , Drosophila melanogaster/metabolismo , Drosophila melanogaster/patogenicidade , Neurônios/metabolismo , Neurônios/patologia
7.
Genes Cells ; 22(1): 105-114, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27868313

RESUMO

Suboptimal nutrition imposes developmental constraints on infant animals, which marshal adaptive responses to eventually become mature adults. Such responses are mounted at multiple levels from systemic to cellular. At the cellular level, the underlying mechanisms of cell proliferation control have been intensively studied. However, less is known about how growth of postmitotic and morphologically complex cells, such as neurons, is controlled by nutritional status. We address this question using Class I and Class IV dendritic arborization neurons in Drosophila larvae. Class IV neurons have been shown to sense nociceptive thermal, mechanical and light stimuli, whereas Class I neurons are proprioceptors. We reared larvae on diets with different protein and carbohydrate content throughout larval stages and examined how morphologies of Class I or Class IV neurons were affected. Dendritic arbors of Class IV neurons became more complex when larvae were reared on a low-yeast diet, which contains lower amounts of amino acids and other ingredients, compared to a high-yeast diet. In contrast, such low-yeast-dependent hyperarborization was not seen in Class I neurons. The physiological and metabolic implications of the hyperarborization phenotype are discussed in relation to a recent hypothesis that Class IV neurons sense protein-deficient stress and to our characterization of how the dietary yeast contents impacted larval metabolism.


Assuntos
Dendritos/genética , Drosophila melanogaster/genética , Larva/genética , Neurônios/metabolismo , Animais , Carboidratos/administração & dosagem , Proliferação de Células/genética , Dendritos/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Plasticidade Neuronal , Neurônios/classificação , Estado Nutricional/genética , Proteínas/administração & dosagem , Células Receptoras Sensoriais/metabolismo
8.
Elife ; 52016 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-26880554

RESUMO

Adequate responses to noxious stimuli causing tissue damages are essential for organismal survival. Class IV neurons in Drosophila larvae are polymodal nociceptors responsible for thermal, mechanical, and light sensation. Importantly, activation of Class IV provoked distinct avoidance behaviors, depending on the inputs. We found that noxious thermal stimuli, but not blue light stimulation, caused a unique pattern of Class IV, which were composed of pauses after high-frequency spike trains and a large Ca(2+) rise in the dendrite (the Ca(2+) transient). Both these responses depended on two TRPA channels and the L-type voltage-gated calcium channel (L-VGCC), showing that the thermosensation provokes Ca(2+) influx. The precipitous fluctuation of firing rate in Class IV neurons enhanced the robust heat avoidance. We hypothesize that the Ca(2+) influx can be a key signal encoding a specific modality.


Assuntos
Cálcio/metabolismo , Dendritos/metabolismo , Dendritos/efeitos da radiação , Drosophila/efeitos da radiação , Temperatura Alta , Nociceptores/efeitos da radiação , Potenciais de Ação , Animais , Canais de Cálcio/metabolismo , Cátions Bivalentes/metabolismo , Drosophila/fisiologia , Nociceptores/fisiologia , Canais de Potencial de Receptor Transitório/metabolismo
9.
Sci Rep ; 4: 4415, 2014 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-24643112

RESUMO

Most organs scale proportionally with body size through regulation of individual cell size and/or cell number. Here we addressed how postmitotic and morphologically complex cells such as neurons scale with the body size by using the dendritic arbor of one Drosophila sensory neuron as an assay system. In small adults eclosed under a limited-nutrition condition, the wild-type neuron preserved the branching complexity of the arbor, but scaled down the entire arbor, making a "miniature". In contrast, mutant neurons for the Insulin/IGF signaling (IIS) or TORC1 pathway exhibited "undergrowth", which was characterized by decreases in both the branching complexity and the arbor size, despite a normal diet. These contrasting phenotypes hinted that a novel regulatory mechanism contributes to the dendritic scaling in wild-type neurons. Indeed, we isolated a mutation in the gene CHORD/morgana that uncoupled the neuron size and the body size: CHORD mutant neurons generated miniature dendritic arbors regardless of the body size. CHORD encodes an evolutionarily conserved co-chaperone of HSP90. Our results support the notion that dendritic growth and branching are controlled by partly separate mechanisms. The IIS/TORC1 pathways control both growth and branching to avert underdevelopment, whereas CHORD together with TORC2 realizes proportional scaling of the entire arbor.


Assuntos
Proteínas de Transporte/metabolismo , Sequência Conservada , Dendritos/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Chaperonas Moleculares/metabolismo , Células Receptoras Sensoriais/metabolismo , Sequência de Aminoácidos , Animais , Tamanho Corporal , Proteínas de Transporte/genética , Tamanho Celular , Dendritos/ultraestrutura , Proteínas de Drosophila/genética , Drosophila melanogaster/anatomia & histologia , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Evolução Molecular , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Insulina/genética , Insulina/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina , Chaperonas Moleculares/genética , Dados de Sequência Molecular , Complexos Multiproteicos/genética , Complexos Multiproteicos/metabolismo , Células Receptoras Sensoriais/ultraestrutura , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
10.
Dev Cell ; 27(5): 530-44, 2013 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-24290980

RESUMO

The transcription factors Abrupt (Ab) and Knot (Kn) act as selectors of distinct dendritic arbor morphologies in two classes of Drosophila sensory neurons, termed class I and class IV, respectively. We performed binding-site mapping and transcriptional profiling of these isolated neurons. Their profiles were similarly enriched in cell-type-specific enhancers of genes implicated in neural development. We identified a total of 429 target genes, of which 56 were common to Ab and Kn; these targets included genes necessary to shape dendritic arbors in either or both of the two sensory subtypes. Furthermore, a common target gene, encoding the cell adhesion molecule Ten-m, was expressed more strongly in class I than class IV, and this differential was critical to the class-selective directional control of dendritic branch sprouting or extension. Our analyses illustrate how differentiating neurons employ distinct and shared repertoires of gene expression to produce class-selective morphological traits.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/fisiologia , Morfogênese/genética , Proteínas Nucleares/genética , Células Receptoras Sensoriais/fisiologia , Fatores de Transcrição/genética , Transcrição Gênica/fisiologia , Animais , Cromatina/genética , Dendritos/fisiologia , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Estudo de Associação Genômica Ampla , Larva/citologia , Larva/fisiologia , Proteínas Nucleares/metabolismo , Células Receptoras Sensoriais/ultraestrutura , Tenascina/genética , Tenascina/metabolismo , Fatores de Transcrição/metabolismo , Transcriptoma
11.
J Cell Sci ; 125(Pt 8): 1980-90, 2012 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-22328496

RESUMO

Septate junctions (SJs) are the membrane specializations observed between epithelial cells in invertebrates. SJs play a crucial role in epithelial barrier function by restricting the free diffusion of solutes through the intercellular space. In arthropod species, two morphologically different types of SJs have been described: pleated septate junctions (pSJs) and smooth septate junctions (sSJs), which are specific to ectodermal and endodermal epithelia, respectively. In contrast to the recent identification of pSJ-related proteins, the molecular constituents of sSJs are mostly unknown. Here, we report the discovery of a new sSJ-specific membrane protein, designated 'Snakeskin' (Ssk). Ssk is highly concentrated in sSJs in the Drosophila midgut and Malpighian tubules. Lack of Ssk expression is embryonically lethal in Drosophila and results in defective sSJ formation accompanied by abnormal morphology of midgut epithelial cells. We also show that the barrier function of the midgut to a fluorescent tracer is impaired in ssk-knockdown larvae. These results suggest that Ssk is required for the intestinal barrier function in Drosophila.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila/metabolismo , Intestinos/citologia , Proteínas de Membrana/metabolismo , Junções Íntimas/metabolismo , Sequência de Aminoácidos , Animais , Drosophila/química , Drosophila/embriologia , Drosophila/genética , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Ectoderma/embriologia , Ectoderma/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/embriologia , Proteínas de Membrana/química , Proteínas de Membrana/genética , Dados de Sequência Molecular , Alinhamento de Sequência , Junções Íntimas/química , Junções Íntimas/genética
12.
Genes Dev ; 25(18): 1982-96, 2011 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-21937715

RESUMO

Members of the Flamingo cadherin family are required in a number of different in vivo contexts of neural development. Even so, molecular identities downstream from the family have been poorly understood. Here we show that a LIM domain protein, Espinas (Esn), binds to an intracellular juxtamembrane domain of Flamingo (Fmi), and that this Fmi-Esn interplay elicits repulsion between dendritic branches of Drosophila sensory neurons. In wild-type larvae, branches of the same class IV dendritic arborization neuron achieve efficient coverage of its two-dimensional receptive field with minimum overlap with each other. However, this self-avoidance was disrupted in a fmi hypomorphic mutant, in an esn knockout homozygote, and in the fmi/esn trans-heterozygote. A functional fusion protein, Fmi:3eGFP, was localized at most of the branch tips, and in a heterologous system, assembly of Esn at cell contact sites required its LIM domain and Fmi. We further show that genes controlling epithelial planar cell polarity (PCP), such as Van Gogh (Vang) and RhoA, are also necessary for the self-avoidance, and that fmi genetically interacts with these loci. On the basis of these and other results, we propose that the Fmi-Esn complex, together with the PCP regulators and the Tricornered (Trc) signaling pathway, executes the repulsive interaction between isoneuronal dendritic branches.


Assuntos
Caderinas/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Drosophila melanogaster/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Animais , Dendritos/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Inativação de Genes , Proteínas do Tecido Nervoso/genética , Sistema Nervoso/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Células Receptoras Sensoriais/citologia , Células Receptoras Sensoriais/metabolismo
13.
Nat Neurosci ; 14(3): 314-23, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21317905

RESUMO

Neuronal connections are often organized in layers that contain synapses between neurons that have similar functions. In Drosophila, R7 and R8 photoreceptors, which detect different wavelengths, form synapses in distinct medulla layers. The mechanisms underlying the specificity of synaptic-layer selection remain unclear. We found that Golden Goal (Gogo) and Flamingo (Fmi), two cell-surface proteins involved in photoreceptor targeting, functionally interact in R8 photoreceptor axons. Our results indicate that Gogo promotes R8 photoreceptor axon adhesion to the temporary layer M1, whereas Gogo and Fmi collaborate to mediate axon targeting to the final layer M3. Structure-function analysis suggested that Gogo and Fmi interact with intracellular components through the Gogo cytoplasmic domain. Moreover, Fmi was also required in target cells for R8 photoreceptor axon targeting. We propose that Gogo acts as a functional partner of Fmi for R8 photoreceptor axon targeting and that the dynamic regulation of their interaction specifies synaptic-layer selection of photoreceptors.


Assuntos
Caderinas/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/anatomia & histologia , Drosophila melanogaster/fisiologia , Proteínas do Olho/metabolismo , Células Fotorreceptoras de Invertebrados/fisiologia , Receptores de Superfície Celular/metabolismo , Sinapses/metabolismo , Animais , Animais Geneticamente Modificados , Axônios/metabolismo , Axônios/ultraestrutura , Caderinas/genética , Proteínas de Drosophila/genética , Proteínas do Olho/genética , Vias Neurais/anatomia & histologia , Vias Neurais/fisiologia , Fenótipo , Células Fotorreceptoras de Invertebrados/ultraestrutura , Receptores de Superfície Celular/genética
14.
Neural Dev ; 4: 37, 2009 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-19799768

RESUMO

BACKGROUND: For the establishment of functional neural circuits that support a wide range of animal behaviors, initial circuits formed in early development have to be reorganized. One way to achieve this is local remodeling of the circuitry hardwiring. To genetically investigate the underlying mechanisms of this remodeling, one model system employs a major group of Drosophila multidendritic sensory neurons - the dendritic arborization (da) neurons - which exhibit dramatic dendritic pruning and subsequent growth during metamorphosis. The 15 da neurons are identified in each larval abdominal hemisegment and are classified into four categories - classes I to IV - in order of increasing size of their receptive fields and/or arbor complexity at the mature larval stage. Our knowledge regarding the anatomy and developmental basis of adult da neurons is still fragmentary. RESULTS: We identified multidendritic neurons in the adult Drosophila abdomen, visualized the dendritic arbors of the individual neurons, and traced the origins of those cells back to the larval stage. There were six da neurons in abdominal hemisegment 3 or 4 (A3/4) of the pharate adult and the adult just after eclosion, five of which were persistent larval da neurons. We quantitatively analyzed dendritic arbors of three of the six adult neurons and examined expression in the pharate adult of key transcription factors that result in the larval class-selective dendritic morphologies. The 'baseline design' of A3/4 in the adult was further modified in a segment-dependent and age-dependent manner. One of our notable findings is that a larval class I neuron, ddaE, completed dendritic remodeling in A2 to A4 and then underwent caspase-dependent cell death within 1 week after eclosion, while homologous neurons in A5 and in more posterior segments degenerated at pupal stages. Another finding is that the dendritic arbor of a class IV neuron, v'ada, was immediately reshaped during post-eclosion growth. It exhibited prominent radial-to-lattice transformation in 1-day-old adults, and the resultant lattice-shaped arbor persisted throughout adult life. CONCLUSION: Our study provides the basis on which we can investigate the genetic programs controlling dendritic remodeling and programmed cell death of adult neurons, and the life-long maintenance of dendritic arbors.


Assuntos
Dendritos/fisiologia , Células Receptoras Sensoriais/fisiologia , Abdome/crescimento & desenvolvimento , Abdome/inervação , Abdome/fisiologia , Envelhecimento/fisiologia , Animais , Animais Geneticamente Modificados , Apoptose/fisiologia , Caspases/metabolismo , Morte Celular/fisiologia , Drosophila , Feminino , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Imuno-Histoquímica , Larva/fisiologia , Masculino , Pupa/fisiologia , Células Receptoras Sensoriais/citologia , Fatores de Tempo
15.
Kansenshogaku Zasshi ; 82(5): 434-40, 2008 Sep.
Artigo em Japonês | MEDLINE | ID: mdl-18975586

RESUMO

We studied 96 mass outbreaks of infectious gastroenteritis due to Norovirus in winter 2006-2007. Of these, 56 occurred in welfare institutions for aging adults 31 in hospitals, and 9 in other facilities such as kinder gardens. Affected staff accounted for 25.9% and users (inpatients, etc.) for 74.1%. The shortest outbreak lasted 10 days and the longest 67 days. We found a positive corelation between periods from the beginning of an outbreak to warnings by public health centers and periods from the beginning to the end of outbreaks. The sooner advice was aired by public health centers, the sooner outbreaks ended. Dementia among users and insufficient knowledge and skills of staff were high risk factors in outbreaks. All 74 of specimens which we examined showed the GII4 genotype. We observed reoutbreaks at three institutions. We compared first and second specimens from the same institution. Two specimens from the second outbreak belonged to the same cluster as the first outbreak. We analyzed 310 bases of RT-PCR products in Capsid regions in both specimens, finding three point mutations accompanied by amino acids changes. This may change the antigenicity of Capsid protein, and may be why reoutbreaks occurred so quickly.


Assuntos
Infecções por Caliciviridae/epidemiologia , Surtos de Doenças , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Idoso , Idoso de 80 Anos ou mais , Infecções por Caliciviridae/virologia , Gastroenterite/virologia , Genótipo , Instituição de Longa Permanência para Idosos/estatística & dados numéricos , Humanos , Japão/epidemiologia , Pessoa de Meia-Idade , Norovirus/genética , Mutação Puntual , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo
17.
J Cell Sci ; 119(Pt 6): 1118-29, 2006 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-16507587

RESUMO

Seven-pass transmembrane cadherins (7-TM cadherins) play pleiotropic roles in epithelial planar cell polarity, shaping dendritic arbors and in axonal outgrowth. In contrast to their role in planar polarity, how 7-TM cadherins control dendritic and axonal outgrowth at the molecular level is largely unknown. Therefore, we performed extensive structure-function analysis of the Drosophila 7-TM cadherin Flamingo (Fmi) and investigated the activities of individual mutant forms mostly in dendritogenesis of dendritic arborization (da) neurons. One of the fmi-mutant phenotypes was overgrowth of branches in the early stage of dendrite development. In da neurons but not in their adjacent non-neuronal cells, expression of a truncated form (deltaN) that lacks the entire cadherin repeat sequence, rescues flies--at least partially--from this phenotype. Another phenotype is observed at a later stage, when dendritic terminals outgrowing from the contralateral sides meet and then avoid each other. In the fmi mutant, by contrast, those branches overlapped. Overexpression of the deltaN form on the wild-type background phenocopied the overlap phenotype in the mutant, and analysis in heterologous systems supported the possibility that this effect might be because the Fmi-Fmi homophilic interaction is inhibited by deltaN. We propose that a dual molecular function of Fmi play pivotal roles in dendrite morphogenesis. In the initial growing phase, Fmi might function as a receptor for a sofar-unidentified ligand and this hypothetical heterophilic interaction would be responsible for limiting branch elongation. At a later stage, homophilic Fmi-binding at dendro-dendritic interfaces would elicit avoidance between dendritic terminals.


Assuntos
Caderinas , Dendritos/química , Proteínas de Drosophila , Neurônios/química , Animais , Caderinas/química , Caderinas/metabolismo , Polaridade Celular , Dendritos/metabolismo , Dendritos/ultraestrutura , Drosophila/embriologia , Proteínas de Drosophila/química , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Modelos Químicos , Morfogênese , Mutação , Neurônios/citologia , Neurônios/fisiologia , Relação Estrutura-Atividade
19.
Curr Biol ; 13(10): 828-32, 2003 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-12747830

RESUMO

Photoreceptors (R cells) in the Drosophila retina connect to targets in three distinct layers of the optic lobe of the brain: R1-R6 connect to the lamina, and R7 and R8 connect to distinct layers in the medulla. In each of these layers, R axon termini are arranged in evenly spaced topographic arrays. In a genetic screen for mutants with abnormal R cell connectivity, we recovered mutations in flamingo (fmi). fmi encodes a seven-transmembrane cadherin, previously shown to function in planar cell polarity and in dendritic patterning. Here, we show that fmi has two specific functions in R8 axon targeting: it facilitates competitive interactions between adjacent R8 axons to ensure their correct spacing, and it promotes the formation of stable connections between R8 axons and their target cells in the medulla. The former suggests a general role for Fmi in establishing nonoverlapping dendritic and axonal target fields. The latter, together with the finding that N-Cadherin has an analogous role in R7 axon-target interactions, points to a cadherin-based system for target layer specificity in the Drosophila visual system.


Assuntos
Axônios/fisiologia , Caderinas/metabolismo , Drosophila/fisiologia , Células Fotorreceptoras de Invertebrados/metabolismo , Vias Visuais/metabolismo , Alelos , Animais , Axônios/ultraestrutura , Encéfalo/metabolismo , Encéfalo/ultraestrutura , Caderinas/genética , Drosophila/metabolismo , Drosophila/ultraestrutura , Proteínas de Drosophila , Regulação da Expressão Gênica/fisiologia , Cones de Crescimento/metabolismo , Cones de Crescimento/ultraestrutura , Heterozigoto , Larva/crescimento & desenvolvimento , Mosaicismo , Mutação , Células Fotorreceptoras de Invertebrados/crescimento & desenvolvimento , Células Fotorreceptoras de Invertebrados/ultraestrutura , Pupa/crescimento & desenvolvimento , Retina/fisiologia , Retina/ultraestrutura , Vias Visuais/crescimento & desenvolvimento , Vias Visuais/ultraestrutura
20.
Development ; 130(6): 1203-13, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12571111

RESUMO

Neurons undergo extensive morphogenesis during development. To systematically identify genes important for different aspects of neuronal morphogenesis, we performed a genetic screen using the MARCM system in the mushroom body (MB) neurons of the Drosophila brain. Mutations on the right arm of chromosome 2 (which contains approximately 20% of the Drosophila genome) were made homozygous in a small subset of uniquely labeled MB neurons. Independently mutagenized chromosomes (4600) were screened, yielding defects in neuroblast proliferation, cell size, membrane trafficking, and axon and dendrite morphogenesis. We report mutations that affect these different aspects of morphogenesis and phenotypically characterize a subset. We found that roadblock, which encodes a dynein light chain, exhibits reduced cell number in neuroblast clones, reduced dendritic complexity and defective axonal transport. These phenotypes are nearly identical to mutations in dynein heavy chain Dhc64 and in Lis1, the Drosophila homolog of human lissencephaly 1, reinforcing the role of the dynein complex in cell proliferation, dendritic morphogenesis and axonal transport. Phenotypic analysis of short stop/kakapo, which encodes a large cytoskeletal linker protein, reveals a novel function in regulating microtubule polarity in neurons. MB neurons mutant for flamingo, which encodes a seven transmembrane cadherin, extend processes beyond their wild-type dendritic territories. Overexpression of Flamingo results in axon retraction. Our results suggest that most genes involved in neuronal morphogenesis play multiple roles in different aspects of neural development, rather than performing a dedicated function limited to a specific process.


Assuntos
Drosophila/embriologia , Drosophila/genética , Corpos Pedunculados/embriologia , Neurônios/metabolismo , Animais , Axônios/metabolismo , Caderinas/genética , Caderinas/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Dendritos/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Mutação
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