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1

Imaging method of neuronal nuclei in the mouse brain

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Czaban L., Broszkiewicz M., Magalska A., Ruszczycki B., Wilczynski G. M.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr Suppl.1
Little is known about the relationship between neuronal gene expression and the architecture of the neuronal cell nucleus. We wanted to study the cell nuclei in neurons of the mouse cerebral cortex, using confocal microscopic immunocytochemistry. We used a segmentation algorithm, based on continuous boundary tracing, able to reconstruct the nucleus surface and to separate adjacent nuclei (Walczak et al. 2013). The algorithm did not use a rigid threshold what made it robust against variations in image intensity and poor contrast. However, when we analyzed mouse, but not rat, neuronal nuclei there have occurred a considerable problem with an appropriate segmentation. This problem was related to the presence of the discrete chromocenters, which are much more prominent in the mouse that in other species. Therefore, in order to assure the proper segmentation, we used sections co-immunostained for the lamin protein. Our refined program is an efficient segmentation tool for crowded and overlapping objects in 3D space, regardless of the particular species. It allows us to study quantitatively the architecture of the neuronal nucleus using confocal-microscopic approach.
2

Aneuploidy and cancer; Bcr-Abl expression leads to compromised mitotic spindle checkpoint promoting chromosomal instability

86%
Piwocka K., Wolanin K., Magalska A., Vejda S., McKenna S.-L., Sikora E.
Acta Biochimica Polonica
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2007
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tom 54
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nr Supplement 4
3
Content available remote

Ciprofloxacin inhibits proliferation and promotes generation of aneuploidy in Jurkat cells

86%
Koziel R., Szczepanowska J., Magalska A., Piwocka K., Duszynski J., Zablocki K.
Journal of Physiology and Pharmacology
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2010
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tom 61
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nr 2
Ciprofloxacin is widely used in antimicrobial therapy. However it also inhibits mitochondrial topoisomerase II and therefore affects cellular energy metabolism. At a concentration exceeding 80 µg/ml ciprofloxacin induces apoptosis, while at 25 µg/ml it inhibits proliferation of Jurkat cells without any symptoms of cell death. The aim of this study was to explain the mechanisms of ciprofloxacin-evoked perturbations of the cell cycle. Human lymphoidal cells (Jurkat) were exposed to ciprofloxacin (25 µg/ml) for 4-11 days and effects of the drug on cell proliferation (light microscopy), cell cycle (flow cytometry), cell size and morphology (confocal microscopy) as well as number of chromosomes (chromosomal spread analysis) were investigated. Exposition of Jurkat cells to ciprofloxacin inhibited cell proliferation, increased proportion of cells in the G2/M-phase of the cell cycle, compromised formation of the mitotic spindle and induced aneuploidy. These observations indicate that ciprofloxacin applied at concentrations insufficient for induction of apoptosis may stop cell proliferation by inhibition of mitosis. Chromosomal instability of such cells may, at least potentially, increase a risk of cancer development.
4

Irregular nuclear shapes of neural stem cells in adult murine dentate gyrus

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Grymanowska A. W., Martin A., Olszynski K. H., Wilczynski G. M., Magalska A., Filipkowski R. K.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: In adult hippocampal neurogenesis, stem cells and their derivative progenitors are generated. They differentiate into neurons as they migrate from the subgranular zone to the granule cell layer of the dentate gyrus, maintaining homeostatic tissue regeneration and supporting brain plasticity. Depending on the stage of the neurogenesis, different subpopulations of cells of the neurogenic lineage can be distinguished, i.e. neural stem cells (NSC, type 1), intermediate progenitor cells (type 2a and type 2b), neuroblasts (type 3) and granule neurons (type 4). Little is known about the architecture of nuclei in these cells, while the cell nucleus is known to be highly organized with numerous functional and structural domains as well as dynamic organization of chromatin governed by epigenetic mechanisms which were shown to respond to external signals. AIM(S): We aimed to distinguish type 1 through 4 cells and investigate their nuclear shape. METHOD(S): Transgenic Nestin-GFP mice were used. Cell types were identified with immunohistochemistry and morphological features: type 1 (GFP+, one long neural process), type 2a (GFP+), type 2b (GFP+/DCX+), type 3 (DCX+), type 4 (NeuN+). Confocal microscopy was used to collect z‑stack files of the nuclei of different cell types. RESULTS: We observed irregularity in shape of the nuclei in type 1 cells (NSC) with the presence of nuclear envelope invaginations. When selected layers were analyzed, NSC nuclei turned out to have reduced circularity, roundness and solidity when compared with other cell types. CONCLUSIONS: The irregularity observed and nuclear envelope invaginations seem to be characteristics of the “stemness” as the shape of the nucleus becomes more regular with successive stages of neurogenesis. The biological significance of the observed phenomenon is not yet clear and further studies are necessary to better understand the process of adult neurogenesis at the nuclear level. FINANCIAL SUPPORT: The work was supported by National Science Centre, Poland, grant no. 2014/14/M/NZ4/00561.
5

Curcumin induces cell death without oligonucleosomal DNA fragmentation in quiescent and proliferating human CD8+ cells

86%
Magalska A., Brzezinska A., Bielak-Zmijewska A., Piwocka K., Mosieniak G., Sikora E.
Acta Biochimica Polonica
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2006
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tom 53
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nr 3
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