DIC, differential interference contrast EGF, epidermal growth factor. ( A) Image sequence of an MDA-MB-231 breast cancer cell that exhibited multiple NE ruptures while moving through 2- × 5-μm 2 constrictions. S2) and could also be detected by accumulation of the fluorescently labeled DNA-binding proteins’ barrier-to-autointegration factor (BAF) ( 15) and guanosine 3′,5′-monophosphate–adenosine 3′,5′-monophosphate (cyclic GMP-AMP) synthase (cGAS) ( 16) at sites of NE rupture (fig. NE rupture was associated with transient influx of fluorescently labeled cytoplasmic proteins into the nucleus (fig.
Piel de marta animal skin#
Breast cancer, fibrosarcoma, and human skin fibroblast cells displayed transient loss of NE integrity, which coincided with the nucleus passing through the constrictions ( Fig.
We detected NE rupture using previously established fluorescent reporters consisting of green or red fluorescent proteins fused to a nuclear localization sequence (NLS-GFP and NLS-RFP, respectively) that rapidly escape into the cytoplasm when NE integrity is lost ( 12– 14). To model cancer cell invasion with precise control over cell confinement, we designed a microfluidic device containing constrictions with fixed height and varying widths mimicking interstitial pore sizes (fig. Thus, we investigated whether cell migration through confining spaces induces NE rupture and compromises DNA integrity and how cells repair such NE ruptures during interphase. We hypothesized that migration through such tight spaces provides a substantial mechanical challenge to the integrity of the nucleus. Whereas the cytoplasm of migrating cells can penetrate even submicron-sized pores, the deformation of the large and relatively rigid nucleus becomes a rate-limiting factor in migration through pores <25 μm 2 in cross section ( 4, 6– 10). In cancer progression, key steps of tumor cell invasion depend upon deformation of the nucleus into available spaces within the three-dimensional tissue ( 3– 6). Loss of NE integrity and nuclear pore selectivity has been linked to the normal aging process and a variety of human diseases, including cancer ( 2). Therefore it is suggested that fibroblasts could work as host cells for parasite survival and permanency in the infected animals.The nuclear envelope (NE), comprising the inner and outer nuclear membranes, nuclear pore complexes, and the nuclear lamina, presents a physical barrier between the nuclear interior and the cytoplasm that protects the genome from cytoplasmic components and establishes a separate compartment for DNA and RNA synthesis and processing ( 1). On the other hand, presence of amastigote multiplication inside of parasitophorus vacuole, showed by electronic microscopy images, probes a true parasite transformation. These results suggest that the parasite affinity for each animal, as well as any intracellular environment resistance, could involve genetic factors in the parasite multiplication. Parasite development in hamsters and mice fibroblasts was evident but there was not multiplication in rat cells showing that apparently they are refractory to Leishmania infection. In addition multiplication rate differences in the cells from these rodent species were determined, for L. This process was demonstrated either by counting parasites inside the stained cells or by electronic microscopy studies. mexicana inside of skin fibroblasts from hamsters, mice and rats was achieved.
Infection and multiplication of Leishmania infantum and L.
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