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Cytogenetic instability of chromosomal nucleolar organizer regions (NORs) in cloned mouse L929 fibroblasts

  • Olga V. Zatsepina
Original Article
  • 25 Downloads

Abstract

Ribosomal DNA (rDNA) gene codes for 18S, 5.8S, and 28S rRNA form tandem repetitive clusters, which occupy distinct chromosomal loci called nucleolar organizer regions (NORs). The number and position of NORs on chromosomes are genetic characteristics of the species although within a cell, the NOR sizes can significantly vary due to loss or multiplication of rDNA copies. In the current study, we used mouse L929 fibroblasts, the aneuploid cells which differ in the FISH- and Ag-NOR numbers, to examine whether the parental NOR variability is inherited in clones. By statistical analysis, we showed that the cloned fibroblasts were able to restore the NOR numerical characteristics of the parental cells after long-term culturing. These results support the idea that mammalian cells may have mechanisms which control the number and activity of NORs at the population level. In L929 fibroblasts, we also regularly observed laterally asymmetry of FISH-NORs that evidenced in an unequal distribution of the mother rDNA copies between the daughter cells in mitosis.

Keywords

mouse L929 fibroblasts clones nucleolar organizer regions rDNA-FISH Ag-NOR staining variability 

Abbreviations

NOR

Nucleolar organizer region

rDNA

Ribosomal DNA

rRNA

Ribosomal RNA

5′ETS

5′ External transcribed spacer

ITS1 and ITS2

The first and second internal transcribed spacers

FISH

Fluorescence in situ hybridization

FISH-NOR

NOR hybridized with rDNA-FISH probes

Ag-NOR

Silver-positive NOR

Notes

Acknowledgements

The author is grateful to Prof. Ingrid Grummt (German Cancer Research Center, Heidelberg, Germany) for a kind provision of the rDNA plasmids, and to Drs. Maria Chaplina, Iana Riabukha, and Anastasiya Moraleva (Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia) for a valuable assistance in some experiments (MC) and statistical evaluation of the selected data (IR, AM).

Author contribution

The author designed the study, participated in the majority of experiments, performed a statistical analysis, wrote the paper, and prepared the illustrations.

Funding

The study was supported by the Russian Foundation for Basic Research (grant 16-04-01199).

Supplementary material

10577_2018_9598_MOESM1_ESM.xlsx (141 kb)
ESM 1 (XLSX 141 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Shemyakin–Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussian Federation

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