Artigo
Leishmania amazonensis Promastigotes Present Two Distinct Modes of Nucleus and Kinetoplast Segregation during Cell Cycle
Registro en:
Plos One. San Francisco: Public Library Science, v. 8, n. 11, 12 p., 2013.
1932-6203
10.1371/journal.pone.0081397
WOS:000327539800128
WOS000327539800128.pdf
7449821021440644
Autor
Silva, Marcelo Santos da [UNESP]
Monteiro, Jomar Patricio
Nunes, Vinicius Santana [UNESP]
Vasconcelos, Elton Jose
Perez, Arina Marina [UNESP]
Freitas-Junior, Lucio de Holanda
Elias, Maria Carolina
Nogueira Cano, Maria Isabel [UNESP]
Resumen
Here, we show the morphological events associated with organelle segregation and their timing in the cell cycle of a reference strain of Leishmania (L.) amazonensis promastigotes, the main causative agent of Tegumentary leishmaniasis in the Americas. We show evidences that during the cell cycle, L. amazonensis promastigotes present two distinct modes of nucleus and kinetoplast segregation, which occur in different temporal order in different proportions of cells. We used DAPI-staining and EdU-labeling to monitor the segregation of DNA-containing organelles and DNA replication in wild-type parasites. The emergence of a new flagellum was observed using a specific monoclonal antibody. The results show that L. amazonensis cell cycle division is peculiar, with 65% of the dividing cells duplicating the kinetoplast before the nucleus, and the remaining 35% doing the opposite or duplicating both organelles concomitantly. In both cases, the new flagellum appeared during S to G2 phase in 1N1K cells and thus before the segregation of both DNA-containing organelles; however, we could not determine the exact timing of flagellar synthesis. Most of these results were confirmed by the synchronization of parasites using hydroxyurea. Altogether, our data show that during the cell cycle of L. amazonensis promastigotes, similarly to L. donovani, the segregation of nucleus and kinetoplast do not follow a specific order, especially when compared to other trypanosomatids, reinforcing the idea that this characteristic seems to be species-specific and may represent differences in cellular biology among members of the Leishmania genus. Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Univ Estadual Paulista UNESP, Inst Biociencias, Dept Genet, Sao Paulo, Brazil Univ Estadual Campinas UNICAMP, Sao Paulo, Brazil Empresa Brasileira Pesquisa Agr Embrapa Caprinos, Sobral, Ceara, Brazil Seattle Biomed Res Inst, Seattle, WA 98109 USA Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias LNBio, Sao Paulo, Brazil Inst Butantan, Ctr Toxins Immune Response & Cell Signaling CeTIC, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Inst Biociencias, Dept Genet, Sao Paulo, Brazil FAPESP: 12/50263-5 CNPq: 4744252010-0 CNPq: PDE 202223/2012-4