How do caveolae help cells adapt to mechanical stress in regeneration and development?
Ian Swinburne, Professor
Molecular and Cell Biology
Open. Apprentices needed for the fall semester. Enter your application online beginning August 21st. The deadline to apply is Monday, August 31st, 4pm.
Mechanical stress is a fact of life. Fluid pressure is required for the function of the circulatory system, ear, and eye. Tissues stretch and contract during morphogenesis, locomotion, and wound healing. But how do tissues (and the cells that constitute them) adapt and respond to changes in the physical stresses they experience?
One potential mechanism is through caveolae, which are specialized plasma membrane invaginations found in many vertebrate tissues subjected to stress, including the skin, muscle, notochord, and blood vessels. Caveolae have been proposed to serve as mechanosensitive membrane buffers and signaling hubs, as caveolae (typically bulb-shaped) have been observed to “flatten” under mechanical stress. This flattening both increases available membrane while enabling new signaling interactions by releasing caveolar accessory proteins to the cytoplasm and allowing previously sequestered membrane proteins to diffuse. However, much of our knowledge of caveolae comes from studies in cultured cells, leaving questions about how these mechanosensitive proteins operate in tissue-scale dynamics. To enable live, in vivo imaging of caveolar dynamics, our lab has recently endogenously tagged caveolar proteins in zebrafish using photostable fluorescent proteins and is beginning to investigate the roles they play in regulating tissue tension in regeneration and development.
Wounded skin serves as a model for studying caveolae in a tissue that experiences stress through both cell stretching and contraction, which occurs during the healing process. Immediately after wounding, we have already observed dramatic relocalization of membrane protein caveolin-1 in the epidermis using amputation and laser ablation assays. New transgenic lines will enable us to visualize and quantify both caveolar formation and flattening in this process for the first time, and we are beginning to mechanistically investigate the role caveolar proteins play in regeneration through genetic perturbations and precisely generated partial loss-of-function mutants. In development, we are also using the yolk sac as a model epithelium and modulating yolk volume in real time to observe how caveolae adapt to changing tissue tension. Based on a recently hypothesized model that links caveolar function to promoting lipid homeostasis, we are also generating transgenic biosensors to characterize whether local lipid concentrations drive caveolar formation or dissociation in these contexts. Collectively, these investigations will characterize caveolae in novel contexts and deepen our mechanistic understanding of its involvement in mechanoadaptive processes that help protect tissues from catastrophic failure in development and regeneration.
We are looking to recruit up to two URAP students for this project. The project scope will be tailored to each student’s individual interests and will be discussed during the interview stage. Apprentices will join and participate in the Swinburne Lab’s diverse scientific community, with the possibility of being offered extensions to future semesters!
Role: URAP apprentice projects will be tailored to individual students' interests, but generally will involve some combination of:
● Confocal and light-sheet microscopy to characterize the responses of biosensors and endogenously tagged proteins in wound healing and development
● Wound healing/regeneration assays in zebrafish
● Live imaging of zebrafish embryos
● Creation of transgenic zebrafish lines (endogenous tags, biosensor integration, and loss-of-function mutants) by leveraging CRISPR systems
● Embryo microinjections for transgenesis, mRNA expression, and genetic perturbations
● Zebrafish husbandry (crossing, embryo collection, screening, and genotyping)
● 4-D image analysis and data visualization
● Critically reading and analyzing papers from relevant scientific literature
● Scientific communication and presentation skills
Qualifications: ● We are seeking students who are passionate about furthering scientific discovery through research and who have an interest in some combination of developmental and/or regenerative biology, cell biology, and biophysics!
● There is no specific background required for this position, but strong candidates will be able to clearly articulate why they are interested in research and their favorite area(s) of science and should be planning on taking advanced coursework in at least one of the listed areas at some point in their undergraduate career.
● An ideal candidate is detail-oriented, highly motivated, passionate about scientific inquiry, and has a clear vision for how a URAP experience will support them in furthering their career goals.
● Candidates should be proactive problem solvers who are not afraid to take initiative and share scientific ideas, questions, and hypotheses in a collaborative and supportive lab environment as they work towards independence in research.
Day-to-day supervisor for this project: Kyle Vallone, Graduate Student
Hours: 12 or more hours
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Biological & Health Sciences