ABSTRACT
Plasma constructions including genes encoding C. trachomatis inclusion membrane protein as composite proteins with reporter green fluorescent protein were obtained. After transfection of HeLa cell culture with the resultant plasmid constructions the location of inclusion membrane proteins in transfected cell was for the first time detected by confocal microscopy.
Subject(s)
Bacterial Proteins , Chlamydia trachomatis , Membrane Proteins , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Chlamydia trachomatis/genetics , Chlamydia trachomatis/metabolism , Gene Expression , HeLa Cells , Humans , Membrane Proteins/genetics , Membrane Proteins/metabolism , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolismABSTRACT
In mouse zygotes, ribosomal genes (rDNA) are transcriptionally silent and so-called "nucleolar precursor bodies" are present instead of typical nucleoli. However, the functional significance of these structures remains obscure. Specifically, it remains unknown whether structural association between the nucleolar precursor bodies and rDNA are maintained when rDNA synthesis is switched off. Here, we studied for the first time the rDNA topology in one-cell mouse embryos and MII oocytes using fluorescence in situ hybridization and mouse rDNA probes. Our data suggest that in the pronuclei of one-cell embryos, rDNAs form rather compact clusters, whose number does not exceed that of nucleolus organizing chromosomes characteristic for the haploid set of mouse chromosomes. In zygotic pronuclei, not all nucleolar precursor bodies are associated with rDNA and not all rDNA repeats are attached to the nucleolar precursor bodies. Altogether, these data favor the idea that spatial interactions of nucleolus organizing chromosomes and nucleolar precursor bodies are not obligatory. We assume that associations between nucleolar precursor bodies and nucleolus organizing chromosomal regions are mediated by centromeric heterochromatin. The total numbers of silver stained nucleolus organizing chromosomes in CBA and C57BL mice are different. rDNA genes are unequally distributed among nucleolus organizing chromosomes and nucleolus organizing regions of sister chromatids.
Subject(s)
Chromosomes, Mammalian/ultrastructure , DNA, Ribosomal/metabolism , Embryo, Mammalian/ultrastructure , Nucleolus Organizer Region/genetics , Oocytes/ultrastructure , Animals , Cell Nucleolus/genetics , Cell Nucleolus/metabolism , Chromosomes, Mammalian/genetics , Embryo, Mammalian/metabolism , Female , In Situ Hybridization, Fluorescence , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Nucleolus Organizer Region/metabolism , Oocytes/metabolismABSTRACT
Peripheral blood platelets of 25 normal subjects were examined by optic microscopy and flow cytometry. Blood was collected routinely into tubes for Cobas Micros flow scintillator with EDTA anticoagulant. Smears were made as a monolayer and stained by May-Grünwald's method in our modification. Morphologically, 5 classes of platelets were distinguished under optic microscope: young, mature, old, macroplatelets, and irritated forms. The percentage of each class was estimated (i.e., normal platelet formula was determined). Comparison of platelet parameters obtained on a flow cytometer and optic microscopy data for subjects with high percentage of large platelets showed increased mean volume of platelets (MPV) and width of platelet volume (PDW). The curve reflecting platelet distribution by volume in these donors was characterized by strong inclination to the right and a flat plateau.
