Microscopy methods for the study of centriole biogenesis and function in Drosophila.
Centrosomes regulate cell motility, adhesion, and polarity in interphase and participate in spindle formation in mitosis. They are composed of two centrioles, which are microtubule-based structures, and a proteinaceous matrix recruited by those, called pericentriolar material. Centrioles are also necessary for the nucleation of the axoneme, the microtubule inner structure of cilia and flagella. The fruit fly, Drosophila melanogaster, has played an important role in the study of cell biology processes and their contextualization in a variety of developmental phenomena. In this chapter, we describe immunofluorescence and electron microscopy methods used to study Drosophila early embryogenesis and spermatogenesis. These methods have been widely used to study centriole assembly and its function as a centrosome organizer during mitotic and meiotic cell divisions and as an axoneme nucleator in the formation of flagella.
Martins
Ana Rodrigues
AR
Instituto Gulbenkian de Ciência, Rua da Quinta Grande, P-2780-156 Oeiras, Portugal.
Machado
Pedro
P
Callaini
Giuliano
G
Bettencourt-Dias
Monica
M
D000818:Animals; D002502:Centrioles / Q000502:physiology*; D004331:Drosophila melanogaster / Q000196:embryology* / Q000502:physiology* / Q000648:ultrastructure; D004625:Embryo, Nonmammalian; D008297:Male; D008853:Microscopy / Q000379:methods*; D008954:Models, Biological; D013091:Spermatogenesis / Q000502:physiology