Changes in lamina structure are followed by spatial reorganization of heterochromatic regions in caspase-8-activated human mesenchymal stem cells

Abstract

Apoptosis is fundamental to the regulation of homeostasis of stem cells in vivo. Whereas the pathways underlying the molecular and biochemical details of nuclear breakdown that accompanies apoptosis have been elucidated, the precise nature of nuclear reorganization that precedes the demolition phase is not fully understood. Here, we expressed an inducible caspase-8 in human mesenchymal stem cells, and quantitatively followed the early changes in nuclear organization during apoptosis. We found that caspase-8 induces alteration of the nuclear lamina and a subsequent spatial reorganization of both centromeres, which are shifted towards a peripheral localization, and telomeres, which form aggregates. This nuclear reorganization correlates with caspase-3 sensitivity of lamina proteins, because the expression of lamin mutant constructs with caspase-3 hypersensitivity resulted in a caspase-8-independent appearance of lamina intranuclear structures and telomere aggregates, whereas application of a caspase inhibitor restrains these changes in nuclear reorganization. Notably, upon activation of apoptosis, we observed no initial changes in the spatial organization of the promyelocytic leukemia nuclear bodies (PML-NBs). We suggest that during activation of the caspase-8 pathway changes in the lamina structure precede changes in heterochromatin spatial organization, and the subsequent breakdown of lamina and PML-NB.

Caspase‐2L, caspase‐9, and caspase‐3 during in vitro maturation and fragmentation of the mouse oocyte

“…Caspase-3 is also known to cleave Nup153, a protein of the nuclear pore complex, and thus to be involved in nuclear envelope breakdown (Buendia et al, 1999), and caspase-3 was also observed along the nuclear envelope of the oocytes. Moreover, the nuclear reorganization that precedes the demolition phase correlates with caspase-3 sensitivity of lamina proteins (Raz et al, 2006).…”

Modeling Oculopharyngeal Muscular Dystrophy in Myotube Cultures Reveals Reduced Accumulation of Soluble Mutant PABPN1 Protein

“…36 Fluorescence in situ hybridization was performed using biotinylated 2=-O-alkyl polyuridine, as previously described. 10 Images were recorded with a Zeiss Axiovert, Oberkochen, Germany (model 135TV) fluorescence microscope equipped with a 100-W mercury arc lamp and a ϫ40/1.3 NA plan Apo objective.…”

Nuclear deformability and telomere dynamics are regulated by cell geometric constraints

“…Cytoskeletal to nuclear links are also essential to the maintenance of poised euchromatin and more repressive condensed chromatin, i.e., heterochromatin assembly (44). Heterochromatin is stabilized by links between the actin cytoskeleton and the nuclear membrane (45)(46)(47). In addition, telomeres, the ends of chromosomes, are necessary for genome stability because they protect chromosomes from DNA damage response machinery (48).…”