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XB-ART-57955
Exp Cell Res March 3, 2021; 401 (2): 112523.
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Characterization of axolotl lampbrush chromosomes by fluorescence in situ hybridization and immunostaining.

Keinath MC , Davidian A , Timoshevskiy V , Timoshevskaya N , Gall JG .


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.

PubMed ID: 33675804
PMC ID: PMC8123938
Article link: Exp Cell Res


Genes referenced: akap13 chd4 cldn7 dicer1 etv4 fgf9 jarid2 klf4 nipbl nisch tex2.1 tmx2 trmt5


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References [+] :
Altschul, Basic local alignment search tool. 1990, Pubmed