DAPI staining and DNA content estimation of nuclei in uncultivable microbial eukaryotes (Arcellinida and Ciliates).

Ciliophora / genetics DNA Indoles Lobosea Phylogeny Staining and Labeling
Arcellinida Ciliates DAPI staining DNA content Nucleus Protists

Journal

European journal of protistology
ISSN: 1618-0429
Titre abrégé: Eur J Protistol
Pays: Germany
ID NLM: 8917383

Informations de publication

Date de publication:
Oct 2021
Historique:
received: 23 02 2021
revised: 09 08 2021
accepted: 30 08 2021
pubmed: 31 10 2021
medline: 12 11 2021
entrez: 30 10 2021
Statut: ppublish

Résumé

Though representing a major component of eukaryotic biodiversity, many microbial eukaryotes remain poorly studied, including the focus of the present work, testate amoebae of the order Arcellinida (Amoebozoa) and non-model lineages of ciliates (Alveolata). In particular, knowledge of genome structures and changes in genome content over the often-complex life cycles of these lineages remains enigmatic. However, the limited available knowledge suggests that microbial eukaryotes have the potential to challenge our textbook views on eukaryotic genomes and genome evolution. In this study, we developed protocols for DAPI (4',6-diamidino-2-phenylindole) staining of Arcellinida nuclei and adapted protocols for ciliates. In addition, image analysis software was used to estimate the DNA content in the nuclei of Arcellinida and ciliates, and the measurements of target organisms were compared to those  of well-known model organisms.The results demonstrate that the methods we have developed for nuclear staining in these lineages are effective and can be applied to other microbial eukaryotic groups by adjusting certain stages in the protocols.

Identifiants

DOI: 10.1016/j.ejop.2021.125840 PMID: 34717075 PMC: PMC8699166
pubmed: 34717075
pii: S0932-4739(21)00075-4
doi: 10.1016/j.ejop.2021.125840
pmc: PMC8699166
mid: NIHMS1740244
pii:
doi:

Substances chimiques

Indoles 0
DAPI 47165-04-8
DNA 9007-49-2

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

125840

Subventions

Organisme : NHGRI NIH HHS
ID : R15 HG010409
Pays : United States

Informations de copyright

Copyright © 2021 Elsevier GmbH. All rights reserved.

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Auteurs

Ketty Munyenyembe (K)

Smith College, Department of Biological Sciences, Northampton, MA, USA.

Caitlin Timmons (C)

Smith College, Department of Biological Sciences, Northampton, MA, USA.

Agnes K M Weiner (AKM)

Smith College, Department of Biological Sciences, Northampton, MA, USA.

Laura A Katz (LA)

Smith College, Department of Biological Sciences, Northampton, MA, USA; University of Massachusetts Amherst, Program in Organismic and Evolutionary Biology, Amherst, MA, USA. Electronic address: lkatz@smith.edu.

Ying Yan (Y)

Smith College, Department of Biological Sciences, Northampton, MA, USA. Electronic address: yingyan@ouc.edu.cn.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Alveolata › Ciliophora › DAPI staining and DNA content estimation of...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ADN › DAPI staining and DNA content estimation of...
questionsmedicales.fr › Composés hétérocycliques › Composés hétérocycliques à cycles fusionnés › Composés hétérobicycliques › Indoles › DAPI staining and DNA content estimation of...
questionsmedicales.fr › Eucaryotes › Amoebozoa › Lobosea › DAPI staining and DNA content estimation of...
questionsmedicales.fr › Sciences de l'information › Phylogenèse › DAPI staining and DNA content estimation of...
questionsmedicales.fr › Techniques d'investigation › Techniques de laboratoire clinique › Techniques histologiques › Techniques de préparation histocytologique › Coloration et marquage › DAPI staining and DNA content estimation of...