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1.
Methods Mol Biol ; 2626: 399-444, 2023.
Article in English | MEDLINE | ID: mdl-36715918

ABSTRACT

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.


Subject(s)
Drosophila , Public Health , Animals , Research
2.
STAR Protoc ; 2(2): 100592, 2021 06 18.
Article in English | MEDLINE | ID: mdl-34169286

ABSTRACT

We have outlined the approach of visualizing autophagy specifically in the epithelial follicle stem cells of the Drosophila ovary using the LysoTracker dye. The advantage of using this protocol is that it details several techniques, including ovary dissection, immunofluorescence, and western blotting, that positively identify autophagy changes in a very small population of cells. One of the limitations of this protocol is that it needs to be combined with other genetic manipulations and positive markers of the autophagy pathway. For complete details on the use and execution of this protocol, please refer to Singh et al., (2018).


Subject(s)
Autophagy , Cell Tracking , Ovarian Follicle/cytology , Stem Cells/cytology , Animals , Drosophila , Epithelial Cells/cytology , Female , Microscopy, Confocal
3.
Dev Cell ; 46(6): 720-734.e6, 2018 09 24.
Article in English | MEDLINE | ID: mdl-30197240

ABSTRACT

Egg production declines with age in many species, a process linked with stem cell loss. Diet-dependent signaling has emerged as critical for stem cell maintenance during aging. Follicle stem cells (FSCs) in the Drosophila ovary are exquisitely responsive to diet-induced signals including Hedgehog (Hh) and insulin-IGF signaling (IIS), entering quiescence in the absence of nutrients and initiating proliferation rapidly upon feeding. Although highly proliferative FSCs generally exhibit an extended lifespan, we find that constitutive Hh signaling drives FSC loss and premature sterility despite high proliferative rates. This occurs due to Hh-mediated induction of autophagy in FSCs via a Ptc-dependent, Smo-independent mechanism. Hh-dependent autophagy increases during aging, triggering FSC loss and consequent reproductive arrest. IIS is necessary and sufficient to suppress Hh-induced autophagy, promoting a stable proliferative state. These results suggest that opposing action of diet-responsive IIS and Hh signals determine reproductive lifespan by modulating the proliferation-autophagy balance in FSCs during aging.


Subject(s)
Autophagy , Cell Proliferation , Drosophila Proteins/metabolism , Drosophila melanogaster/physiology , Hedgehog Proteins/metabolism , Insulin/pharmacology , Ovarian Follicle/cytology , Stem Cells/cytology , Animals , Drosophila Proteins/genetics , Drosophila melanogaster/drug effects , Female , Hedgehog Proteins/genetics , Hypoglycemic Agents/pharmacology , Insulin-Like Growth Factor I/genetics , Insulin-Like Growth Factor I/metabolism , Male , Ovarian Follicle/drug effects , Ovarian Follicle/metabolism , Receptors, Cell Surface/genetics , Receptors, Cell Surface/metabolism , Signal Transduction/drug effects , Smoothened Receptor/genetics , Smoothened Receptor/metabolism , Stem Cell Niche/drug effects , Stem Cells/drug effects , Stem Cells/metabolism
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