Abstract
The nematode Caenorhabditis elegans is one of the major model organisms in cell and developmental biology. This organism is easy to culture in laboratories and suitable for microscopic investigation of the cytoskeleton. Because the worms are small and transparent, the actin cytoskeleton in many tissues and cells can be observed with appropriate visualization techniques without sectioning or dissection. This chapter describes the introduction to representative methods for imaging the actin cytoskeleton in C. elegans and a protocol for staining worms with fluorescent phalloidin.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Brenner S (1974) The genetics of Caenorhabditis elegans. Genetics 77:71–94
White JG, Southgate E, Thomson JN, Brenner S (1986) The structure of the nervous system of the nematode Caenorhabditis elegans. Philos Trans R Soc Lond Ser B Biol Sci 314:1–340
Sulston JE, Schierenberg E, White JG, Thomson JN (1983) The embryonic cell lineage of the nematode Caenorhabditis elegans. Dev Biol 100:64–119
Sherwood DR, Plastino J (2018) Invading, leading and navigating cells in Caenorhabditis elegans: insights into cell movement in vivo. Genetics 208:53–78
Pintard L, Bowerman B (2019) Mitotic cell division in Caenorhabditis elegans. Genetics 211:35–73
Goldstein B, Nance J (2020) Caenorhabditis elegans gastrulation: a model for understanding how cells polarize change shape and journey toward the center of an embryo. Genetics 214:265–277
Ono S (2014) Regulation of structure and function of sarcomeric actin filaments in striated muscle of the nematode Caenorhabditis elegans. Anat Rec 297:1548–1559
Benian GM, Epstein HF (2011) Caenorhabditis elegans muscle: a genetic and molecular model for protein interactions in the heart. Circ Res 109:1082–1095
Chisholm AD, Hutter H, Jin Y, Wadsworth WG (2016) The genetics of axon guidance and axon regeneration in Caenorhabditis elegans. Genetics 204:849–882
Harris H, Tso M, Epstein HF (1977) Actin and myosin-linked calcium regulation in the nematode Caenorhabditis elegans: biochemical and structural properties of native filaments and purified proteins. Biochemistry 16:859–865
Ono S (1999) Purification and biochemical characterization of actin from Caenorhabditis elegans: its difference from rabbit muscle actin in the interaction with nematode ADF/cofilin. Cell Motil Cytoskeleton 43:128–136
Ono S, Pruyne D (2012) Biochemical and cell biological analysis of actin in the nematode Caenorhabditis elegans. Methods 56:11–17
Ono S, Baillie DL, Benian GM (1999) UNC-60B an ADF/cofilin family protein is required for proper assembly of actin into myofibrils in Caenorhabditis elegans body wall muscle. J Cell Biol 145:491–502
Ono K, Parast M, Alberico C, Benian GM, Ono S (2003) Specific requirement for two ADF/cofilin isoforms in distinct actin-dependent processes in Caenorhabditis elegans. J Cell Sci 116:2073–2085
Mohri K, Ono K, Yu R, Yamashiro S, Ono S (2006) Enhancement of actin-depolymerizing factor/cofilin-dependent actin disassembly by actin-interacting protein 1 is required for organized actin filament assembly in the Caenorhabditis elegans body wall muscle. Mol Biol Cell 17:2190–2199
Duerr JS (2006) Immunohistochemistry WormBook ed the C elegans research community WormBook. PubMed PMID: 18050446; PMCID: PMC4780882; https://doi.org/10.1895/wormbook.1.105.1
Finney M, Ruvkun G (1990) The unc-86 gene product couples cell lineage and cell identity in C elegans. Cell 63:895–905
Shakes DC, Miller DM, Nonet ML (2012) Immunofluorescence microscopy. Methods Cell Biol 107:35–66
Wilson KJ, Qadota H, Benian GM (2012) Immunofluorescent localization of proteins in Caenorhabditis elegans muscle. Methods Mol Biol 798:171–181
Riedl J, Crevenna AH, Kessenbrock K, Yu JH, Neukirchen D, Bista M, Bradke F, Jenne D, Holak TA, Werb Z, Sixt M, Wedlich-Soldner R (2008) Lifeact: a versatile marker to visualize F-actin. Nat Methods 5:605–607
Burkel BM, von Dassow G, Bement WM (2007) Versatile fluorescent probes for actin filaments based on the actin-binding domain of utrophin. Cell Motil Cytoskeleton 64:822–832
Edwards KA, Demsky M, Montague RA, Weymouth N, Kiehart DP (1997) GFP-moesin illuminates actin cytoskeleton dynamics in living tissue and demonstrates cell shape changes during morphogenesis in drosophila. Dev Biol 191:103–117
Aizawa H, Sameshima M, Yahara I (1997) A green fluorescent protein-actin fusion protein dominantly inhibits cytokinesis cell spreading and locomotion in Dictyostelium. Cell Struct Funct 22:335–345
Willis JH, Munro E, Lyczak R, Bowerman B (2006) Conditional dominant mutations in the Caenorhabditis elegans gene act-2 identify cytoplasmic and muscle roles for a redundant actin isoform. Mol Biol Cell 17:1051–1064
Stone S, Shaw JE (1993) A Caenorhabditis elegans act-4::lacZ fusion: use as a transformation marker and analysis of tissue-specific expression. Gene 131:167–173
MacQueen AJ, Baggett JJ, Perumov N, Bauer RA, Januszewski T, Schriefer L, Waddle JA (2005) ACT-5 is an essential Caenorhabditis elegans actin required for intestinal microvilli formation. Mol Biol Cell 16:3247–3259
Victor Ambros. (2006) Shaham S (ed) WormBook: Methods in Cell Biology (January 02, 2006), WormBook, ed. The C. elegans Research Community, WormBook, https://doi.org/10.1895/wormbook.1.49.1, http://www.wormbook.org.
McCarter J, Bartlett B, Dang T, Schedl T (1997) Soma-germ cell interactions in Caenorhabditis elegans: multiple events of hermaphrodite germline development require the somatic sheath and spermathecal lineages. Dev Biol 181:121–143
Dong L, Cornaglia M, Krishnamani G, Zhang J, Mouchiroud L, Lehnert T, Auwerx J, Gijs MAM (2018) Reversible and long-term immobilization in a hydrogel-microbead matrix for high-resolution imaging of Caenorhabditis elegans and other small organisms. PLoS One 13:e0193989
Hwang H, Barnes E, Matsunaga Y, Benian GM, Ono S, Lu H (2016) Muscle contraction phenotypic analysis enabled by optogenetics reveals functional relationships of sarcomere components in Caenorhabditis elegans. Sci Rep 6:19900
Krajniak J, Lu H (2010) Long-term high-resolution imaging and culture of C elegans in chip-gel hybrid microfluidic device for developmental studies. Lab Chip 10:1862–1868
Lengsfeld AM, Low I, Wieland T, Dancker P, Hasselbach W (1974) Interaction of phalloidin with actin. Proc Natl Acad Sci U S A 71:2803–2807
Wulf E, Deboben A, Bautz FA, Faulstich H, Wieland T (1979) Fluorescent phallotoxin a tool for the visualization of cellular actin. Proc Natl Acad Sci U S A 76:4498–4502
Strome S (1986) Fluorescence visualization of the distribution of microfilaments in gonads and early embryos of the nematode Caenorhabditis elegans. J Cell Biol 103:2241–2252
Waterston RH, Hirsh D, Lane TR (1984) Dominant mutations affecting muscle structure in Caenorhabditis elegans that map near the actin gene cluster. J Mol Biol 180:473–496
Ono S (2001) The Caenorhabditis elegans unc-78 gene encodes a homologue of actin-interacting protein 1 required for organized assembly of muscle actin filaments. J Cell Biol 152:1313–1319
Hayashi Y, Ono K, Ono S (2019) Mutations in Caenorhabditis elegans actin which are equivalent to human cardiomyopathy mutations cause abnormal actin aggregation in nematode striated muscle. F1000Res 8:279
Barnes DE, Hwang H, Ono K, Lu H, Ono S (2016) Molecular evolution of troponin I and a role of its N-terminal extension in nematode locomotion. Cytoskeleton 73:117–130
Stiernagle T (2006) Maintenance of C elegans. WormBook ed The C elegans Research Community, PubMed PMID: 18050451; PMCID: PMC4781397 https://doi.org/10.1895/wormbook.1.101.1
Hall DH, Altun ZF (2008) C elegans atlas. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Lessard JL (1988) Two monoclonal antibodies to actin: one muscle selective and one generally reactive. Cell Motil Cytoskeleton 10:349–362
Porta-de-la-Riva M, Fontrodona L, Villanueva A, Ceron J (2012) Basic Caenorhabditis elegans methods: synchronization and observation. J Vis Exp 10:e4019
Tse YC, Werner M, Longhini KM, Labbe JC, Goldstein B, Glotzer M (2012) RhoA activation during polarization and cytokinesis of the early Caenorhabditis elegans embryo is differentially dependent on NOP-1 and CYK-4. Mol Biol Cell 23:4020–4031
Zilberman Y, Abrams J, Anderson DC, Nance J (2017) Cdc42 regulates junctional actin but not cell polarization in the Caenorhabditis elegans epidermis. J Cell Biol 216:3729–3744
Higuchi-Sanabria R, Paul JW 3rd, Durieux J, Benitez C, Frankino PA, Tronnes SU, Garcia G, Daniele JR, Monshietehadi S, Dillin A (2018) Spatial regulation of the actin cytoskeleton by HSF-1 during aging. Mol Biol Cell 29:2522–2527
Xu S, Chisholm AD (2011) A Gαq-Ca2+ signaling pathway promotes actin-mediated epidermal wound closure in C elegans. Curr Biol 21:1960–1967
Shivas JM, Skop AR (2012) Arp2/3 mediates early endosome dynamics necessary for the maintenance of PAR asymmetry in Caenorhabditis elegans. Mol Biol Cell 23:1917–1927
Schindler AJ, Sherwood DR (2011) The transcription factor HLH-2/E/daughterless regulates anchor cell invasion across basement membrane in C elegans. Dev Biol 357:380–391
Wirshing ACE, Cram EJ (2017) Myosin activity drives actomyosin bundle formation and organization in contractile cells of the Caenorhabditis elegans spermatheca. Mol Biol Cell 28:1937–1949
Szumowski SC, Estes KA, Popovich JJ, Botts MR, Sek G, Troemel ER (2016) Small GTPases promote actin coat formation on microsporidian pathogens traversing the apical membrane of Caenorhabditis elegans intestinal cells. Cell Microbiol 18:30–45
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Ono, S. (2022). Imaging of Actin Cytoskeleton in the Nematode Caenorhabditis elegans. In: Gavin, R.H. (eds) Cytoskeleton . Methods in Molecular Biology, vol 2364. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1661-1_7
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1661-1_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1660-4
Online ISBN: 978-1-0716-1661-1
eBook Packages: Springer Protocols