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Characterization of axolotl lampbrush chromosomes by fluorescence in situ hybridization and immunostaining.

Author
Abstract
:

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.

Year of Publication
:
2021
Journal
:
Experimental cell research
Volume
:
401
Issue
:
2
Number of Pages
:
112523
Date Published
:
2021
ISSN Number
:
0014-4827
URL
:
https://linkinghub.elsevier.com/retrieve/pii/S0014-4827(21)00054-9
DOI
:
10.1016/j.yexcr.2021.112523
Short Title
:
Exp Cell Res
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