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1.
Methods Mol Biol ; 2626: 399-444, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36715918

RESUMEN

Citizen science is a productive approach to include non-scientists in research efforts that impact particular issues or communities. In most cases, scientists at advanced career stages design high-quality, exciting projects that enable citizen contribution, a crowdsourcing process that drives discovery forward and engages communities. The challenges of having citizens design their own research with no or limited training and providing access to laboratory tools, reagents, and supplies have limited citizen science efforts. This leaves the incredible life experiences and immersion of citizens in communities that experience health disparities out of the research equation, thus hampering efforts to address community health needs with a full picture of the challenges that must be addressed. Here, we present a robust and reproducible approach that engages participants from Grade 5 through adult in research focused on defining how diet impacts disease signaling. We leverage the powerful genetics, cell biology, and biochemistry of Drosophila oogenesis to define how nutrients impact phenotypes associated with genetic mutants that are implicated in cancer and diabetes. Participants lead the project design and execution, flipping the top-down hierarchy of the prevailing scientific culture to co-create research projects and infuse the research with cultural and community relevance.


Asunto(s)
Drosophila , Salud Pública , Animales , Investigación
2.
Cell Signal ; 27(6): 1225-36, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25748048

RESUMEN

Direct interaction of α9ß1 integrin with nerve growth factor (NGF) has been previously reported to induce pro-proliferative and pro-survival activities of non-neuronal cells. We investigated participation of p75(NTR) in α9ß1 integrin-dependent cellular response to NGF stimulation. Using selective transfection of glioma cell lines with these receptors, we showed a strong, cation-independent association of α9 integrin subunit with p75(NTR) on the cellular membrane by selective immunoprecipitation experiments. The presence of the α9/p75(NTR) complex increases NGF-dependent cell adhesion, proliferation and migration. Other integrin subunits including ß1 were not found in complex with p75(NTR). FRET analysis indicated that p75(NTR) and α9 integrin subunit are not closely associated through their cytoplasmic domains, most probably because of the molecular interference with other cytoplasmic proteins such as paxillin. Interaction of α9ß1 integrin with another ligand, VCAM-1 was not modulated by the p75(NTR). α9/p75(NTR) complex elevated NGF-dependent activation of MAPK Erk1/2 arty for integrin that may create active complexes with other types of receptors belonging to the TNF superfamily.


Asunto(s)
Proliferación Celular/efectos de los fármacos , Integrinas/metabolismo , Factor de Crecimiento Nervioso/farmacología , Proteínas del Tejido Nervioso/metabolismo , Receptores de Factor de Crecimiento Nervioso/metabolismo , Animales , Adhesión Celular/efectos de los fármacos , Línea Celular Tumoral , Transferencia Resonante de Energía de Fluorescencia , Células HEK293 , Humanos , Inmunohistoquímica , Integrinas/química , Integrinas/genética , Ratones , Microscopía Confocal , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Factor de Crecimiento Nervioso/aislamiento & purificación , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/genética , Paxillin/metabolismo , Unión Proteica , Receptor trkA/metabolismo , Receptores de Factor de Crecimiento Nervioso/química , Receptores de Factor de Crecimiento Nervioso/genética , Molécula 1 de Adhesión Celular Vascular/metabolismo
3.
Genetics ; 199(4): 935-57, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25680813

RESUMEN

In many tissues, the presence of stem cells is inferred by the capacity of the tissue to maintain homeostasis and undergo repair after injury. Isolation of self-renewing cells with the ability to generate the full array of cells within a given tissue strongly supports this idea, but the identification and genetic manipulation of individual stem cells within their niche remain a challenge. Here we present novel methods for marking and genetically altering epithelial follicle stem cells (FSCs) within the Drosophila ovary. Using these new tools, we define a sequential multistep process that comprises transitioning of FSCs from quiescence to proliferation. We further demonstrate that integrins are cell-autonomously required within FSCs to provide directional signals that are necessary at each step of this process. These methods may be used to define precise roles for specific genes in the sequential events that occur during FSC division after a period of quiescence.


Asunto(s)
Drosophila melanogaster/genética , Perfilación de la Expresión Génica/métodos , Genoma de los Insectos , Integrinas/metabolismo , Folículo Ovárico/citología , Células Madre/metabolismo , Animales , Proliferación Celular , Drosophila melanogaster/citología , Femenino , Integrinas/genética , Masculino , Células Madre/citología , Células Madre/fisiología
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