We studied the alterations of dying oocytes in 1-28 days old rats using TUNEL method, immunolocalizations of active caspase 3, lamp1, localization of acid phosphatase, and DAPI staining. All procedures were performed in adjacent sections of each oocyte. In most dying oocytes exist simultaneously features of apoptosis as active caspase 3 and DNA breaks, and a large increase of lamp1 and acid phosphatase characteristic of autophagy. Large clumps of compact chromatin and membrane blebbing were absent. Electron microscope observations demonstrated the presence of small clear vesicles and autophagolysosomes. All these features indicate that a large number of oocytes are eliminated by a process sharing features of apoptosis and autophagy. In dying oocytes of new born rats the markers of apoptosis predominate over those of autophagy. However, fragmentation and apoptotic bodies were not found. These features suggest that in different cytophysiological conditions the processes of cell death may be differently modulated.
The synaptonemal complex (SC) is an evolutionarily conserved structure that mediates synapsis of homologous chromosomes during meiotic prophase I. Previous studies have established that the chromatin of homologous chromosomes is organized in loops that are attached to the lateral elements (LEs) of the SC. The characterization of the genomic sequences associated with LEs of the SC represents an important step toward understanding meiotic chromosome organization and function. To isolate these genomic sequences, we performed chromatin immunoprecipitation assays in rat spermatocytes using an antibody against SYCP3, a major structural component of the LEs of the SC. Our results demonstrated the reproducible and exclusive isolation of repeat deoxyribonucleic acid (DNA) sequences, in particular long interspersed elements, short interspersed elements, long terminal direct repeats, satellite, and simple repeats. The association of these repeat sequences to the LEs of the SC was confirmed by in situ hybridization of meiotic nuclei shown by both light and electron microscopy. Signals were also detected over the chromatin surrounding SCs and in small loops protruding from the lateral elements into the SC central region. We propose that genomic repeat DNA sequences play a key role in anchoring the chromosome to the protein scaffold of the SC.
By light and electron microscope cytochemistry we characterized the interphase nucleus of Lacandonia schismatica, the only known species of the new plant family Lacandoniaceae, whose most peculiar feature is the inverted position of the sexual organs, an aspect never found before among flowering plants. Furthermore, we compare it to Triuris alata, a related species, to Voyria aphylla (a dicotyledon), to Gymnosiphon divaricatus (a monocotyledon) and also to saprophytes. The reticulated chromatin of L schismatica and T alata is similar to that of other monocotyledons. In addition, we describe a unique type of RNP granules in the interchromatin space which are about 32 + 3 nm SD in diameter and occur as huge clusters. They are intermediate in size and spatial distribution between inter-and peri-chromatin granules. We term them 'Lacandonia granules'. The granules were also found in Talata. They are 3 ! +_ 2 nm in diameter. No significant differences in size were observed between them (P > 0.05). Synaptonemallike complexes and ring-shaped structures were seen in interphase nuclei of somatic cells of these species. Coiled and nucleolus-associated bodies, as well as centromeres were also found in these two organisms. On the contrary, V aphylla and G divaricatus display a chromocentric nuclear organization. The nuclear similarities between L schismatica and Talata suggest extremely close phylogenetic relationships between them. chromatin / Lacandonia I plant nucleus / ribonucleoproteins
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