Drosophila S2 Cells as a Model System to Investigate Mitotic Spindle Dynamics, Architecture, and Function

• Tipo: Outra Publicação em Revista Científica Internacional
• Data: 2010

Autores

Moutinho Pereira, S (Autor) Outra; Matos, I (Autor) Outra; Helder Maiato (Autor) FMUP

Revista

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Título: METHODS IN CELL BIOLOGY, VOL 66

Vol. 51

Páginas: 243-257

ISSN: 0091-679X

Editora: ACADEMIC PRESS INC

Indexação

ISI Web of Knowledge - 10 Citações

Scopus - 11 Citações

Outras Informações

ID Authenticus: P-003-C0G

DOI: 10.1016/s0091-679x(10)97014-3

Abstract (EN): In order to perpetuate their genetic content, eukaryotic cells have developed a microtubule-based machine known as the mitotic spindle. Independently of the system studied, mitotic spindles share at least one common characteristic-the dynamic nature of microtubules. This property allows the constant plasticity needed to assemble a bipolar structure, make proper kinetochore-microtubule attachments, segregate chromosomes, and finally disassemble the spindle and reform an interphase microtubule array. Here, we describe a variety of experimental approaches currently used in our laboratory to study microtubule dynamics during mitosis using Drosophila melanogaster S2 cells as a model. By using quantitative live cell imaging microscopy in combination with an advantageous labeling background, we illustrate how several cooperative pathways are used to build functional mitotic spindles. We illustrate different ways of perturbing spindle microtubule dynamics, including pharmacological inhibition and RNA interference of proteins that directly or indirectly impair microtubule dynamics. Additionally, we demonstrate the advantage of using fluorescent speckle microscopy to investigate an intrinsic property of spindle microtubules known as poleward flux. Finally, we developed a set of laser microsurgery-based experiments that allow, with unique spatiotemporal resolution, the study of specific spindle structures (e.g., centrosomes, microtubules, and kinetochores) and their respective roles during mitosis.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 15