Utilização de abelhas adultas em análises de citometria de fluxo

O primeiro trabalho de citometria de fluxo utilizando abelhas da tribo Meliponini, quantificou o genoma de três espécies, as quais apresentaram conteúdo de DNA de 0,43 pg em Scaptotrigona xantotricha, 0,93 pg em Melipona rufiventris e 0,95 pg em Melipona mondury. Posteriormente à determinação do seu...

Nível de Acesso:openAccess
Publication Date:2018
Main Author: Faustino, Alessandra de Oliveira lattes
Orientador/a: Salomão, Tânia Maria Fernandes
Format: Dissertação
Published: Universidade Federal de Viçosa
Áreas de Conhecimento:
Online Access:http://www.locus.ufv.br/handle/123456789/21596
Citação:FAUSTINO, Alessandra de Oliveira. Utilização de abelhas adultas em análises de citometria de fluxo. 2018. 23 f. Dissertação (Mestrado em Biologia Celular e Estrutural) - Universidade Federal de Viçosa, Viçosa. 2018.
Resumo inglês:The first work of flow cytometry using bees from Meliponini tribe quantified the genome of three species, to which they presented DNA content of 0.43 pg in Scaptotrigona xantotricha, 0.93 pg in Melipona rufiventris and 0.95 pg in Melipona mondury. After the determination of its genomic content, S. xantotricha became to be was used as the internal standard in all the studies of meliponine cytometry. Currently, values of just 63 species of bees are available, showing a scarcity of data in this group. One of the reasons for this data shortage may be due to the difficulty of access the larvae and pupae, which were the only ones used, until the moment, in the studies of quantifying the genome of bees. Thus, the search for the use of more accessible material may be a strategy for more species to be analyzed. Therefore, the present study aimed to test different buffers for the quantification of nuclear DNA content in adult bees. For the preparation of the nuclear suspensions, to be analyzed in the flow cytometer, adult bees of the species S. xantotricha and the internal standard Drosophila melanogaster, along with LB01, Galbraith, MgSO4 and TRIS buffers. The histograms obtained had good resolution levels and coefficient of variation of satisfactory, with percentages lower than 5%, ranging from 2.24% with Galbraith buffer to 3.34% with LBO1 buffer. The size of the genome obtained with the use of adults of S. xantotricha values very close or identical to the available data in the literature for the larvae and pupae of the same species. Similarly, the experimental coefficient did not show great variations in relation to the theoretical one (2.39), and the MgSO4 buffer with a ratio of 2.34 was the most distant. In relation to the percentage of debri cell, the samples using the Galbraith, MgSO4 and TRIS buffers were cleaner compared to those using the LB01 buffer. Finally, the results of analyzed parameter in the same day of extraction and 24 hours after extraction were very similar. In front of this fact, we can conclude that the utilization of adult bee for the determination of DNA content is so effective as to use of larvae and pupae, since its result not showed significant statistical differences when compared with the literature data.