Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior

Samuel J. Belfer, Han-Sheng Chuang, Benjamin L. Freedman, Jinzhou Yuan, Michael Norton, Haim H. Bau, David M. Raizen

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Study Objectives: To develop a method, called Caenorhabditis-in-Drop (CiD), encapsulating single worms in aqueous drops, for parallel analysis of behavioral quiescence in C. elegans nematodes. Design: We designed, constructed, and tested a device that houses an array of aqueous droplets laden with individual worms. The droplets are separated and covered by immiscible, biocompatible oil. We modeled gas exchange across the aqueous/oil interface and tested the viability of the encapsulated animals. We studied the behavior of wild-type animals; of animals with a loss of function mutation in the cGMP-dependent protein kinase gene egl-4; of animals with a loss of function mutation in the gene kin-2, which encodes a cAMP-dependent protein kinase A regulatory subunit; of animals with a gain-of-function mutation in the gene acy-1, which encodes an adenylate cyclase; and of animals that express high levels of the EGF protein encoded by lin-3. Measurements and Results: We used CiD to simultaneously monitor the behavior of 24 worms, a nearly 5-fold improvement over the prior best methodology. In support of our gas exchange models, we found that worms remain viable on the chip for 4 days, past the 12-h period needed for observation, but show reduced longevity to that measured on an agar surface. Measurements of duration of lethargus quiescence and total lethargus quiescence showed reduced amounts as well as reduced variability relative to prior methods. There was reduced lethargus quiescence in animals that were mutant for kin-2 and for acy-1, supporting a wake-promoting effect of PKA in C. elegans, but no change in lethargus quiescence in egl-4 mutants. There was increased quiescence in animals that expressed kin-2 in the nervous system or over-expressed EGF. Conclusions: CiD is useful for the analysis of behavioral quiescence during lethargus as well as during the adult stage C. elegans. The method is expandable to parallel simultaneous monitoring of hundreds of animals and for other studies of long-term behavior. Using this method, we were successful in measuring, for the first time, quiescence in kin-2(ce179) and in acy-2(ce2) mutants, which are hyperactive. Our observations also highlight the impact of environmental conditions on quiescent behavior and show that longevity is reduced in CiD in comparison to agar surfaces.

Original languageEnglish
Pages (from-to)689-698
Number of pages10
JournalSleep
Volume36
Issue number5
DOIs
Publication statusPublished - 2013 May 1

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Caenorhabditis
Cyclic AMP-Dependent Protein Kinases
Epidermal Growth Factor
Mutation
Agar
Oils
Gases
Genes
Cyclic GMP-Dependent Protein Kinases
Wild Animals
Adenylyl Cyclases
Nervous System
Observation
Equipment and Supplies

All Science Journal Classification (ASJC) codes

  • Clinical Neurology
  • Physiology (medical)

Cite this

Belfer, S. J., Chuang, H-S., Freedman, B. L., Yuan, J., Norton, M., Bau, H. H., & Raizen, D. M. (2013). Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior. Sleep, 36(5), 689-698. https://doi.org/10.5665/sleep.2628
Belfer, Samuel J. ; Chuang, Han-Sheng ; Freedman, Benjamin L. ; Yuan, Jinzhou ; Norton, Michael ; Bau, Haim H. ; Raizen, David M. / Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior. In: Sleep. 2013 ; Vol. 36, No. 5. pp. 689-698.
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abstract = "Study Objectives: To develop a method, called Caenorhabditis-in-Drop (CiD), encapsulating single worms in aqueous drops, for parallel analysis of behavioral quiescence in C. elegans nematodes. Design: We designed, constructed, and tested a device that houses an array of aqueous droplets laden with individual worms. The droplets are separated and covered by immiscible, biocompatible oil. We modeled gas exchange across the aqueous/oil interface and tested the viability of the encapsulated animals. We studied the behavior of wild-type animals; of animals with a loss of function mutation in the cGMP-dependent protein kinase gene egl-4; of animals with a loss of function mutation in the gene kin-2, which encodes a cAMP-dependent protein kinase A regulatory subunit; of animals with a gain-of-function mutation in the gene acy-1, which encodes an adenylate cyclase; and of animals that express high levels of the EGF protein encoded by lin-3. Measurements and Results: We used CiD to simultaneously monitor the behavior of 24 worms, a nearly 5-fold improvement over the prior best methodology. In support of our gas exchange models, we found that worms remain viable on the chip for 4 days, past the 12-h period needed for observation, but show reduced longevity to that measured on an agar surface. Measurements of duration of lethargus quiescence and total lethargus quiescence showed reduced amounts as well as reduced variability relative to prior methods. There was reduced lethargus quiescence in animals that were mutant for kin-2 and for acy-1, supporting a wake-promoting effect of PKA in C. elegans, but no change in lethargus quiescence in egl-4 mutants. There was increased quiescence in animals that expressed kin-2 in the nervous system or over-expressed EGF. Conclusions: CiD is useful for the analysis of behavioral quiescence during lethargus as well as during the adult stage C. elegans. The method is expandable to parallel simultaneous monitoring of hundreds of animals and for other studies of long-term behavior. Using this method, we were successful in measuring, for the first time, quiescence in kin-2(ce179) and in acy-2(ce2) mutants, which are hyperactive. Our observations also highlight the impact of environmental conditions on quiescent behavior and show that longevity is reduced in CiD in comparison to agar surfaces.",
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Belfer, SJ, Chuang, H-S, Freedman, BL, Yuan, J, Norton, M, Bau, HH & Raizen, DM 2013, 'Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior', Sleep, vol. 36, no. 5, pp. 689-698. https://doi.org/10.5665/sleep.2628

Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior. / Belfer, Samuel J.; Chuang, Han-Sheng; Freedman, Benjamin L.; Yuan, Jinzhou; Norton, Michael; Bau, Haim H.; Raizen, David M.

In: Sleep, Vol. 36, No. 5, 01.05.2013, p. 689-698.

Research output: Contribution to journalArticle

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AU - Belfer, Samuel J.

AU - Chuang, Han-Sheng

AU - Freedman, Benjamin L.

AU - Yuan, Jinzhou

AU - Norton, Michael

AU - Bau, Haim H.

AU - Raizen, David M.

PY - 2013/5/1

Y1 - 2013/5/1

N2 - Study Objectives: To develop a method, called Caenorhabditis-in-Drop (CiD), encapsulating single worms in aqueous drops, for parallel analysis of behavioral quiescence in C. elegans nematodes. Design: We designed, constructed, and tested a device that houses an array of aqueous droplets laden with individual worms. The droplets are separated and covered by immiscible, biocompatible oil. We modeled gas exchange across the aqueous/oil interface and tested the viability of the encapsulated animals. We studied the behavior of wild-type animals; of animals with a loss of function mutation in the cGMP-dependent protein kinase gene egl-4; of animals with a loss of function mutation in the gene kin-2, which encodes a cAMP-dependent protein kinase A regulatory subunit; of animals with a gain-of-function mutation in the gene acy-1, which encodes an adenylate cyclase; and of animals that express high levels of the EGF protein encoded by lin-3. Measurements and Results: We used CiD to simultaneously monitor the behavior of 24 worms, a nearly 5-fold improvement over the prior best methodology. In support of our gas exchange models, we found that worms remain viable on the chip for 4 days, past the 12-h period needed for observation, but show reduced longevity to that measured on an agar surface. Measurements of duration of lethargus quiescence and total lethargus quiescence showed reduced amounts as well as reduced variability relative to prior methods. There was reduced lethargus quiescence in animals that were mutant for kin-2 and for acy-1, supporting a wake-promoting effect of PKA in C. elegans, but no change in lethargus quiescence in egl-4 mutants. There was increased quiescence in animals that expressed kin-2 in the nervous system or over-expressed EGF. Conclusions: CiD is useful for the analysis of behavioral quiescence during lethargus as well as during the adult stage C. elegans. The method is expandable to parallel simultaneous monitoring of hundreds of animals and for other studies of long-term behavior. Using this method, we were successful in measuring, for the first time, quiescence in kin-2(ce179) and in acy-2(ce2) mutants, which are hyperactive. Our observations also highlight the impact of environmental conditions on quiescent behavior and show that longevity is reduced in CiD in comparison to agar surfaces.

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Belfer SJ, Chuang H-S, Freedman BL, Yuan J, Norton M, Bau HH et al. Caenorhabditis-in-drop array for monitoring C. elegans quiescent behavior. Sleep. 2013 May 1;36(5):689-698. https://doi.org/10.5665/sleep.2628