Quantum dot assisted tracking of the intracellular protein Cyclin E in Xenopus laevis embryos.
BACKGROUND
Luminescent semiconductor nanocrystals, also known as quantum dots (QD), possess highly desirable optical properties that account for development of a variety of exciting biomedical techniques. These properties include long-term stability, brightness, narrow emission spectra, size tunable properties and resistance to photobleaching. QD have many promising applications in biology and the list is constantly growing. These applications include DNA or protein tagging for in vitro assays, deep-tissue imaging, fluorescence resonance energy transfer (FRET), and studying dynamics of cell surface receptors, among others. Here we explored the potential of QD-mediated labeling for the purpose of tracking an intracellular protein inside live cells.


RESULTS
We manufactured dihydrolipoic acid (DHLA)-capped CdSe-ZnS core-shell QD, not available commercially, and coupled them to the cell cycle regulatory protein Cyclin E. We then utilized the QD fluorescence capabilities for visualization of Cyclin E trafficking within cells of Xenopus laevis embryos in real time.


CONCLUSIONS
These studies provide "proof-of-concept" for this approach by tracking QD-tagged Cyclin E within cells of developing embryos, before and during an important developmental period, the midblastula transition. Importantly, we show that the attachment of QD to Cyclin E did not disrupt its proper intracellular distribution prior to and during the midblastula transition. The fate of the QD after cyclin E degradation following the midblastula transition remains unknown.
Department of Cell Biology and Physiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, 87131-0001, USA. ybrandt@salud.unm.edu.
Yekaterina I
YI
Brandt
Department of Cell Biology and Physiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, 87131-0001, USA. mitchelltnt@gmail.com.
Therese
T
Mitchell
Center for High Technology Materials, University of New Mexico, 1313 Goddard SE, Albuquerque, New Mexico, 87106-4343, USA. gen@chtm.unm.edu.
Gennady A
GA
Smolyakov
Center for High Technology Materials, University of New Mexico, 1313 Goddard SE, Albuquerque, New Mexico, 87106-4343, USA. osinski@chtm.unm.edu.
Marek
M
Osiński
Department of Cell Biology and Physiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, 87131-0001, USA. rhartley@salud.unm.edu.
Rebecca S
RS
Hartley
D000818:Animals; D019927:Cyclin E / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods; D045663:Quantum Dots / Q000032:analysis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013440:Sulfides; D008063:Thioctic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D014982:Xenopus laevis / Q000196:embryology*; D017967:Zinc Compounds
D019927:Cyclin E; D011994:Recombinant Proteins; D013440:Sulfides; D017967:Zinc Compounds; D049452:Green Fluorescent Proteins; C033223:polyhistidine; D006639:Histidine; D008063:Thioctic Acid; C015144:dihydrolipoic acid; C031238:zinc sulfide