Chromosome Research

, Volume 19, Issue 2, pp 165–182 | Cite as

Nuclei of chicken neurons in tissues and three-dimensional cell cultures are organized into distinct radial zones

  • Doris Berchtold
  • Stephanie Fesser
  • Gesine Bachmann
  • Alexander Kaiser
  • John-Christian Eilert
  • Florian Frohns
  • Nicolas Sadoni
  • Joscha Muck
  • Elisabeth Kremmer
  • Dirk Eick
  • Paul G. Layer
  • Daniele Zink
Article

Abstract

We used chicken retinospheroids (RS) to study the nuclear architecture of vertebrate cells in a three-dimensional (3D) cell culture system. The results showed that the different neuronal cell types of RS displayed an extreme form of radial nuclear organization. Chromatin was arranged into distinct radial zones which became already visible after DAPI staining. The distinct zones were enriched in different chromatin modifications and in different types of chromosomes. Active isoforms of RNA polymerase II were depleted in the outermost zone. Also chromocenters and nucleoli were radially aligned in the nuclear interior. The splicing factor SC35 was enriched at the central zone and did not show the typical speckled pattern of distribution. Evaluation of neuronal and non-neuronal chicken tissues showed that the highly ordered form of radial nuclear organization was also present in neuronal chicken tissues. Furthermore, the data revealed that the neuron-specific nuclear organization was remodeled when cells spread on a flat substrate. Monolayer cultures of a chicken cell line did not show this extreme form of radial organization. Rather, such monolayer cultures displayed features of nuclear organization which have been described before for many different types of monolayer cells. The finding that an extreme form radial nuclear organization, which has not been described before, is present in RS and tissues, but not in cells spread on a flat substrate, suggests that it would be important to complement studies on nuclear architecture performed with monolayer cells by studies on 3D cell culture systems and tissues.

Keywords

nuclear architecture radial nuclear organization 3D cell culture retinospheroid chicken neuron 

Abbreviations

DAPI

4′,6′-diamidino-2′-phenylindole

2D

Two-dimensional

3D

Three-dimensional

E6

Day 6 of embryonic development

FA

Formaldehyde

FISH

Fluorescence in situ hybridization

Η4Κ8ac

Histone H4 acetylated at lysine 8

PBS

Phosphate-buffered saline

PI

Propidium iodide

Pol II Ser 2

RNA polymerase II phosphorylated at serine 2

RS

Retinospheroid

TT

0.2% Tween 20 and 0.2% Triton X-100

Supplementary material

10577_2010_9182_MOESM1_ESM.doc (26 kb)
Figure S1Part of a RS (light optical section) stained with an antibody against vimentin (green), which is a prominent component of the cytoskeleton of Müller glia cells. Nuclei were stained with DAPI (blue). Arrows point to elongated Müller glia cells. Scale bar 50 μm (DOC 25 kb)

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Doris Berchtold
    • 1
  • Stephanie Fesser
    • 1
  • Gesine Bachmann
    • 2
  • Alexander Kaiser
    • 1
  • John-Christian Eilert
    • 3
  • Florian Frohns
    • 2
  • Nicolas Sadoni
    • 1
  • Joscha Muck
    • 1
    • 4
  • Elisabeth Kremmer
    • 5
  • Dirk Eick
    • 6
  • Paul G. Layer
    • 2
  • Daniele Zink
    • 1
    • 4
  1. 1.Department Biologie IILudwig-Maximilians-Universität (LMU) München, BiozentrumPlanegg-MartinsriedGermany
  2. 2.Technische Universität Darmstadt, Fachbereich BiologieDarmstadtGermany
  3. 3.BioImaging ZentrumPlanegg-MartinsriedGermany
  4. 4.Institute of Bioengineering and NanotechnologySingaporeSingapore
  5. 5.Helmholtz Zentrum München, Institut für Molekulare ImmunologieMünchenGermany
  6. 6.Helmholtz Zentrum München, Abteilung für Molekulare Epigenetik, Center of Integrated Protein ScienceMünchenGermany

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