A new approach to map transcription sites at the ultrastructural level.
We describe a new ultrastructural method for locating transcription on ultra-thin sections. The use of anti-DNA/RNA hybrid antibodies provides specific labeling on precise structures of the nuclear compartments of several cell types. All mammalian and plant material studied (HeLa cells, lymphocytes, onion root meristematic cells) showed the same pattern of labeling: fibrillar structures in the interchromatin region and discrete regions of the dense fibrillar component at the periphery of the fibrillar centers in the nucleolus. The specificity of the immunogold labeling was tested by RNAse H digestion and by pre-blocking the antibody with synthetic DNA/RNA hybrids; in both cases no gold particles were observed. This method has considerable advantages compared with current techniques, constituting a very useful tool to map transcriptionally active loci in a variety of cells.
Testillano
P S
PS
Laboratory of Nuclear Organization and Plant Development, Centro de Investigaciones Biológicas, CSIC, Madrid, Spain.
Gorab
E
E
Risueño
M C
MC
D000490:Allium / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure*; D000906:Antibodies / Q000032:analysis; D000918:Antibody Specificity; D002466:Cell Nucleolus / Q000737:chemistry / Q000648:ultrastructure; D002843:Chromatin / Q000737:chemistry / Q000648:ultrastructure; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007399:Interphase; D008214:Lymphocytes / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure*; D008854:Microscopy, Electron; D012313:RNA / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure; D014158:Transcription, Genetic / Q000235:genetics*
D000906:Antibodies; D002843:Chromatin; D012313:RNA; D004247:DNA