Changes of nuclear structure induced by increasing temperatures.
Despite the recent improvement in understanding the higher-order structure of chromatin fibers, the organization of interphase chromosomes in specific nuclear domains emerged only recently and it is still controversial. This study took advantage of an integrated approach using complementary techniques in order to investigate the structure and organization of chromatin in interphase nucleus. Native CHO-K1 cells were progressively heated from 310 K to 410 K and the effects of increasing temperatures on nuclear chromatin were analyzed in situ by means of cytometric and calorimetric techniques. Distribution and organization of chromatin domains were analyzed by Fluorescence microscopy, while the mean condensation of nuclear chromatin was measured by Differential scanning calorimetry. The results show as changes of nuclear structures (envelope and matrix, namely) affect significantly organization and condensation of in situ chromatin. Moreover when volume is modified by an external force (the temperature gradient in our case) we observe significant alterations of chromatin structure. These data are in accordance with the hypothesis of an inverse relationship between nuclear volume and chromatin condensation.
Department of Biophysical Sciences and Technologies M & O, School of Medicine, University of Genova, Italy. vergani@ibf.unige.it
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D000818:Animals; D016466:CHO Cells; D002152:Calorimetry, Differential Scanning; D022003:Cell Nucleus Structures / Q000737:chemistry* / Q000648:ultrastructure; D002843:Chromatin / Q000737:chemistry / Q000648:ultrastructure*; D002942:Circular Dichroism; D006224:Cricetinae; D005456:Fluorescent Dyes / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D007211:Indoles / Q000737:chemistry; D008856:Microscopy, Fluorescence; D013696:Temperature
D002843:Chromatin; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI