TY - JOUR
T1 - Quantitative imaging of sleep behavior in Caenorhabditis elegans and larval Drosophila melanogaster
AU - Churgin, Matthew A.
AU - Szuperak, Milan
AU - Davis, Kristen C.
AU - Raizen, David M.
AU - Fang-Yen, Christopher
AU - Kayser, Matthew S.
N1 - Funding Information:
We thank P. McClanahan for assistance with the method for fabrication of flat WorMotel bases and for recording the video of pharyngeal pumping in the WorMotel. This work was supported by NIH grants K08NS090461 (M.S.K.), R01NS088432 (D.M.R. and C.F.-Y.), and R01NS084835 (C.F.-Y.); the Ellison Medical Foundation (C.F.-Y.); the European Commission Horizon 2020 program (C.F.-Y.); a Burroughs Wellcome Career Award for Medical Scientists (M.S.K.); a March of Dimes Basil O’Connor Scholar Award (M.S.K.); and a Sloan Research Fellowship (M.S.K.). K. Davis is a trainee in the NIH Translational Research Training Program (T32 ES019851, PI: T. Penning, Penn CEET).
Publisher Copyright:
© 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2019/5/1
Y1 - 2019/5/1
N2 - Sleep is nearly universal among animals, yet remains poorly understood. Recent work has leveraged simple model organisms, such as Caenorhabditis elegans and Drosophila melanogaster larvae, to investigate the genetic and neural bases of sleep. However, manual methods of recording sleep behavior in these systems are labor intensive and low in throughput. To address these limitations, we developed methods for quantitative imaging of individual animals cultivated in custom microfabricated multiwell substrates, and used them to elucidate molecular mechanisms underlying sleep. Here, we describe the steps necessary to design, produce, and image these plates, as well as analyze the resulting behavioral data. We also describe approaches for experimentally manipulating sleep. Following these procedures, after ~2 h of experimental preparation, we are able to simultaneously image 24 C. elegans from the second larval stage to adult stages or 20 Drosophila larvae during the second instar life stage at a spatial resolution of 10 or 27 µm, respectively. Although this system has been optimized to measure activity and quiescence in Caenorhabditis larvae and adults and in Drosophila larvae, it can also be used to assess other behaviors over short or long periods. Moreover, with minor modifications, it can be adapted for the behavioral monitoring of a wide range of small animals.
AB - Sleep is nearly universal among animals, yet remains poorly understood. Recent work has leveraged simple model organisms, such as Caenorhabditis elegans and Drosophila melanogaster larvae, to investigate the genetic and neural bases of sleep. However, manual methods of recording sleep behavior in these systems are labor intensive and low in throughput. To address these limitations, we developed methods for quantitative imaging of individual animals cultivated in custom microfabricated multiwell substrates, and used them to elucidate molecular mechanisms underlying sleep. Here, we describe the steps necessary to design, produce, and image these plates, as well as analyze the resulting behavioral data. We also describe approaches for experimentally manipulating sleep. Following these procedures, after ~2 h of experimental preparation, we are able to simultaneously image 24 C. elegans from the second larval stage to adult stages or 20 Drosophila larvae during the second instar life stage at a spatial resolution of 10 or 27 µm, respectively. Although this system has been optimized to measure activity and quiescence in Caenorhabditis larvae and adults and in Drosophila larvae, it can also be used to assess other behaviors over short or long periods. Moreover, with minor modifications, it can be adapted for the behavioral monitoring of a wide range of small animals.
UR - http://www.scopus.com/inward/record.url?scp=85064002413&partnerID=8YFLogxK
U2 - 10.1038/s41596-019-0146-6
DO - 10.1038/s41596-019-0146-6
M3 - Article
C2 - 30953041
AN - SCOPUS:85064002413
VL - 14
SP - 1455
EP - 1488
JO - Nature Protocols
JF - Nature Protocols
SN - 1754-2189
IS - 5
ER -