Characterization of axolotl lampbrush chromosomes by fluorescence in situ hybridization and immunostaining.
Ambystoma mexicanum
Axolotl
BAC FISH
Centromere
Lampbrush chromosomes
Journal
Experimental cell research
ISSN: 1090-2422
Titre abrégé: Exp Cell Res
Pays: United States
ID NLM: 0373226
Informations de publication
Date de publication:
15 04 2021
15 04 2021
Historique:
received:
12
12
2020
revised:
08
02
2021
accepted:
15
02
2021
pubmed:
7
3
2021
medline:
11
9
2021
entrez:
6
3
2021
Statut:
ppublish
Résumé
The lampbrush chromosomes (LBCs) in oocytes of the Mexican axolotl (Ambystoma mexicanum) were identified some time ago by their relative lengths and predicted centromeres, but they have never been associated completely with the mitotic karyotype, linkage maps or genome assembly. We identified 9 of the axolotl LBCs using RNAseq to identify actively transcribed genes and 13 BAC (bacterial artificial clone) probes containing pieces of active genes. Using read coverage analysis to find candidate centromere sequences, we developed a centromere probe that localizes to all 14 centromeres. Measurements of relative LBC arm lengths and polymerase III localization patterns enabled us to identify all LBCs. This study presents a relatively simple and reliable way to identify each axolotl LBC cytologically and to anchor chromosome-length sequences (from the axolotl genome assembly) to the physical LBCs by immunostaining and fluorescence in situ hybridization. Our data will facilitate a more detailed transcription analysis of individual LBC loops.
Identifiants
pubmed: 33675804
pii: S0014-4827(21)00054-9
doi: 10.1016/j.yexcr.2021.112523
pmc: PMC8123938
mid: NIHMS1687729
pii:
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
112523Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM033397
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
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