A laser pointer driven microheater for precise local heating and conditional gene regulation in vivo. Microheater driven gene regulation in zebrafish.
BACKGROUND
Tissue heating has been employed to study a variety of biological processes, including the study of genes that control embryonic development. Conditional regulation of gene expression is a particularly powerful approach for understanding gene function. One popular method for mis-expressing a gene of interest employs heat-inducible heat shock protein (hsp) promoters. Global heat shock of hsp-promoter-containing transgenic animals induces gene expression throughout all tissues, but does not allow for spatial control. Local heating allows for spatial control of hsp-promoter-driven transgenes, but methods for local heating are cumbersome and variably effective.


RESULTS
We describe a simple, highly controllable, and versatile apparatus for heating biological tissue and other materials on the micron-scale. This microheater employs micron-scale fiber optics and uses an inexpensive laser-pointer as a power source. Optical fibers can be pulled on a standard electrode puller to produce tips of varying sizes that can then be used to reliably heat 20-100 mum targets. We demonstrate precise spatiotemporal control of hsp70l:GFP transgene expression in a variety of tissue types in zebrafish embryos and larvae. We also show how this system can be employed as part of a new method for lineage tracing that would greatly facilitate the study of organogenesis and tissue regulation at any time in the life cycle.


CONCLUSIONS
This versatile and simple local heater has broad utility for the study of gene function and for lineage tracing. This system could be used to control hsp-driven gene expression in any organism simply by bringing the fiber optic tip in contact with the tissue of interest. Beyond these uses for the study of gene function, this device has wide-ranging utility in materials science and could easily be adapted for therapeutic purposes in humans.
Placinta
Mike
M
Department of Biology, University of Massachusetts, Amherst, MA 01003, USA. mikeplacinta@gmail.com
Shen
Meng-Chieh
MC
Achermann
Marc
M
Karlstrom
Rolf O
RO
D000818:Animals; D030801:Animals, Genetically Modified; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D050884:HSP72 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response; D006358:Hot Temperature; D007834:Lasers; D015533:Transcriptional Activation; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism
D050884:HSP72 Heat-Shock Proteins; D049452:Green Fluorescent Proteins