Nuclear Fos domains in transcriptionally activated supraoptic nucleus neurons.
This study has analysed by light and electron microscopy immunolocalization the nuclear pattern of distribution of Fos-related proteins in supraotic neurons. Two experimental models of transcriptional activation have been used: sustained, global transcriptional activation, at relatively near physiological conditions, by six days of chronic intermittent salt loading; and superinduction of c-fos gene by this salt loading regime plus cycloheximide treatment for 4 h. In the first condition, the ultrastructural analysis showed a distribution of Fos-like immunoreactivity on the reticular network of dispersed chromatin that extends between the nucleolar surface and the nuclear envelope, whereas the Fos-negative adjacent interchromatin spaces appeared rich in interchromatin granules by using a cytochemical staining for ribonucleoproteins. The nucleolus associated heterochromatin, fibrillar centers of the nucleolus and coiled bodies were free of immunoreactivity. This immunoelectron pattern seems to indicate that active genes containing activator protein-1 and cyclic AMP response element recognition sites are extensively distributed in euchromatin regions and suggests that the Fos-positive nuclear domains correspond to the actively transcribing chromatin regions, at least in supraoptic neurons. It also suggests that these Fos-positive transcription domains are complementary to adjacent ribonucleoprotein-rich interchromatin spaces which are involved in the processing and splicing of pre-messenger RNA. Moreover, the absence of immunoreactivity on the fibrillar centers, the sites of pre-ribosomal RNA synthesis, suggests that the Fos protein complexes are not involved in regulating the expression of ribosomal RNA genes. Following superinduction of c-fos gene by osmotic stimulation plus cycloheximide treatment, a conspicuous Fos-like immunoreactivity was detected in dispersed chromatin regions, whereas the heterochromatin masses, nucleoli and coiled bodies showed no immunoreaction. Moreover, this treatment induced the formation of nuclear "dense bodies" of a fibrillar nature which were free of immunolabelling. Since Fos proteins are known to be short-lived, the expression of these nuclear constituents, under conditions of protein synthesis inhibition induced by the cycloheximide, suggests the stabilization of chromatin-bound Fos complexes or, alternatively, a preferential synthesis of Fos proteins.
Lafarga
M
M
Departamento de Anatomía y Biología Celular, Facultad de Medicina, Universidad de Cantabria, Santander, Spain.
Martinez-Guijarro
F J
FJ
Berciano
M T
MT
Blasco-Ibañez
J M
JM
Andres
M A
MA
Mellström
B
B
Lopez-Garcia
C
C
Naranjo
J R
JR
D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002843:Chromatin / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D022041:Euchromatin; D016762:Genes, fos; D016758:Genes, jun; D007150:Immunohistochemistry; D008297:Male; D008854:Microscopy, Electron; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013495:Supraoptic Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D014158:Transcription, Genetic / Q000187:drug effects
D002843:Chromatin; D022041:Euchromatin; D016760:Proto-Oncogene Proteins c-fos; D003513:Cycloheximide