title: Breaking Caenorhabditis elegans the easy way using the Balch homogenizer: an old tool for a new application.
abstract: The nematode Caenorhabditis elegans is a model organism best known for its powerful genetics. There is an increasing need in the worm community to couple genetics with biochemistry. Isolation of functionally active proteins or nucleic acids without the use of strong oxidizing denaturants or of subcellular compartments from C. elegans has, however, been challenging because of the worms' thick surrounding cuticle. The Balch homogenizer is a tool that has found much use in mammalian cell culture biology. The interchangeable single ball-bearing design of this instrument permits rapid permeabilization, or homogenization, of cells. Here we demonstrate the utility of the Balch homogenizer for studies with C. elegans. We describe procedures for the efficient breakage and homogenization of every larval stage, including dauers, and show that the Balch homogenizer can be used to extract functionally active proteins. Enzymatic assays for catalase and dihydrolipoamide dehydrogenase show that sample preparation using the Balch homogenizer equals or outperforms conventional methods employing boiling, sonication, or Dounce homogenization. We also describe phenol-free techniques for isolation of genomic DNA and RNA. Finally, we used the tool to isolate coupled mitochondria and polysomes. The reusable Balch homogenizer represents a quick and convenient solution for undertaking biochemical studies on C. elegans.
authors.affiliation: Barshop Institute for Longevity and Aging Studies and Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78240, USA.
authors.forename: Shylesh
authors.initials: S
authors.lastname: Bhaskaran
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Butler
authors.forename: Sandra
authors.initials: S
authors.lastname: Becerra
authors.forename: Veronica
authors.initials: V
authors.lastname: Fassio
authors.forename: Milena
authors.initials: M
authors.lastname: Girotti
authors.forename: Shane L
authors.initials: SL
authors.lastname: Rea
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000302:isolation & purification; D002374:Catalase / Q000737:chemistry / Q000302:isolation & purification; D018509:DNA, Helminth / Q000737:chemistry / Q000302:isolation & purification; D008058:Dihydrolipoamide Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification; D004734:Energy Metabolism; D057075:Enzyme Assays; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000378:metabolism; D010063:Ovum / Q000737:chemistry; D010100:Oxygen / Q000378:metabolism; D011132:Polyribosomes / Q000737:chemistry / Q000378:metabolism; D018510:RNA, Helminth / Q000737:chemistry / Q000302:isolation & purification; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods*
chemical_list: D029742:Caenorhabditis elegans Proteins; D018509:DNA, Helminth; D018510:RNA, Helminth; D002374:Catalase; D008058:Dihydrolipoamide Dehydrogenase; D010100:Oxygen