title: Visualization of cleavage furrow proteins in fixed dividing spermatocytes.
abstract: Cytokinesis separates the cytoplasmic organelles and the duplicated genome into two daughter cells at the end of cell division. In animal cell cytokinesis, assembly and constriction of the contractile apparatus must be finely coordinated with plasma membrane remodeling and vesicle trafficking at the cleavage furrow. Accurate control of these events during cell cleavage is a fundamental task in all organisms and is also essential for maintaining ploidy and preventing neoplastic transformation. Drosophila male meiosis provides a well-suited cell system for exploring the molecular mechanisms underlying cytokinesis, combining the powerful tools of Drosophila genetics with unique cytological characteristics. Remarkably the large size of male meiotic cells highly facilitates cytological analysis of cytokinesis. Here we describe the main procedures that we use for fixing and visualizing cleavage furrow proteins in male meiotic cells. Moreover, we detail our protocol to detect protein interactions in fixed dividing spermatocytes by applying in situ proximity ligation assay.
authors.affiliation: Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma Sapienza, Roma, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Frappaolo
authors.affiliation: Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma Sapienza, Roma, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Sechi
authors.affiliation: Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma Sapienza, Roma, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Belloni
authors.affiliation: Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma Sapienza, Roma, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Piergentili
authors.affiliation: Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma Sapienza, Roma, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Giansanti
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000235:genetics / Q000648:ultrastructure; D048749:Cytokinesis / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster; D008297:Male; D008540:Meiosis / Q000235:genetics; D057054:Molecular Imaging / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D013090:Spermatocytes / Q000254:growth & development / Q000648:ultrastructure*; D013091:Spermatogenesis / Q000235:genetics; D008941:Spindle Apparatus / Q000235:genetics / Q000648:ultrastructure
chemical_list: D029721:Drosophila Proteins
keywords: Contractile ring; Cytokinesis; Drosophila; GOLPH3; Immunofluorescence; In situ protein interaction; Male meiosis; Vesicle trafficking