Induction Of C-fos Protein Research Articles

C-fos protein like immunoreactivity in non-neuronal cells of rat peripheral nerve after transection

The induction of c-fos protein (Fos) in non-neuronal cells of rat sciatic nerve was studied immunocytochemically after transection. As soon as 1 h after transection a few Fos-positive Schwann cells and fibroblast-like cells were seen around the wound margins. In the epi-perineurial fibroblast-like cells Fos-like immunoreactivity (Fos-LI) was most intense at 3 h postoperation, but no Fos-LI was seen at 6 days postoperation. In Schwann cells Fos-LI was spread throughout the distal nerve segment at 6 days postoperation, when the staining (in these cells) was most intense. Fos-immunoreactive Schwann cells in the distal nerve segment were still detectable at 14 days postoperation. In the proximal nerve segment positively labelled Schwann cells were observed only 1 and 3 h postoperation and were localized near the site of injury. The results show that the loss of nerve fibers induces a long-lasting expression of Fos or Fos-related antigens in a peripheral nerve.

Focal ischemia in rats causes time-dependent expression of c-fos protein immunoreactivity in widespread regions of ipsilateral cortex

c-Fos protein expression was examined in brain by immunohistochemistry following permanent middle cerebral artery (MCA) occlusion above the rhinal fissure and ipsilateral common carotid artery (CCA) occlusion in Long-Evans rats. In sham-operated animals, c-fos protein-like immunoreactivity (CFPLI) was confined to neuronal nuclei of the hypothalamus and was not present in other regions including cerebral cortex. In the core territory of the MCA, CFPLI was not detected when examined at 15 and 30 min, 1, 4 and 8 h and 1, 2, 4 and 7 days after occlusion. Focal ischemia induced two temporal and spatial patterns of CFPLI. At 1 h, c-fos protein was expressed in the nuclei of many neurons in layers II–V of the ipsilateral cortex both immediately adjacent to and remote from the ischemic territory. Within regions outside the MCA territory (e.g. cingulate gyrus and prinform cortices), CFPLI in these neurons peaked at 2–4 and was undetectable after 2 days. Neurons in the zone immediately surrounding the ischemic core within MCA territory also expressed CFPLI, but in contrast, continued to express c-fos up to 4 days after ischemia. Immunoreactivity surrounding the ischemic core was found in neuronal nuclei predominantly, although from 1 to 4 days, CFPLI was found in perikarya and dendrites as well. MK-801 (3 mg/kg, i.p., 30 min prior to occlusion) completely blocked the early c-fos protein induction in all regions but expression within neurons surrounding the ischemic core was present 1 day after a single injection. The initial induction of c-fos protein in cortical neurons distant from the ischemic core lesion and prolonged c-fos expression surrounding the infarct suggests that c-fos expression is linked to both early onset-short lasting stimulation and more persistent excitatory or plasticity-related phenomena associated with focal ischemic brain injury.

Global ischemia induces NMDA receptor-mediated c-fos expression in neurons resistant to injury in gerbil hippocampus

The induction of c-fos protein-like immunoreactivity (CFPLI) was examined in the hippocampus of gerbils at several time points after transient global ischemia. c-Fos protein induction was largely confined to the dentate gyrus, CA3 and CA4 regions from 2 to 8 h after transient bilateral carotid occlusion. Little CFPLI was seen in the CA1 subfield, which is disproportionately sensitive to injury after global ischemia. c-Fos induction was completely blocked by pretreatment with MK-801 (3 mg/kg). Our results show that c-fos expression after global ischemia is NMDA receptor mediated, and mainly found in hippocampal neurons resistant to ischemic injury.

Immobilization stress and osmotic stimulation induce c-fos protein in the paraventricular and supraoptic nuclei

Immobilization stress and osmotic stimulation induce c-fos protein in the paraventricular and supraoptic nuclei

Induction of c-fos mRNA and protein in neurons and glia after traumatic brain injury: Pharmacological characterization

Focal brain injury in mice induced c-fos mRNA and protein in neurons throughout the damaged neocortex, including the piriform and the entorhinal cortices, as well as in nonneural brain cells (e.g., glia, pia, ependyma). The pattern of c-fos induction after injury suggested that injury led to spreading depression which then led to c-fos induction in neurons. Human neurons in the temporal cortex and hippocampus also showed c-fos protein induction after neurosurgical trauma. The c-fos mRNA and protein induction in mouse neurons was prevented by the noncompetitive NMDA antagonist ketamine but only partially inhibited by the voltage-dependent calcium channel antagonist nifedipine and the calmodulin antagonist trifluoperazine. The c-fos protein induction in nonneural brain cells after injury was not affected by these drugs. Thus, induction of c-fos in neocortical neurons after focal brain injury is partly NMDA receptor mediated.

MK-801 induces c-fos protein in thalamic and neocortical neurons of rat brain

MK-801, a non-competitive N- methyl- d-aspartate (NMDA) receptor antagonist, leads to a dramatic induction of c-fos-like protein in neurons in deep layers of the neocortex, in dorsal and ventral midline thalamic nuclei and in neurons in the central grey of rat brain. This induction of c-fos by MK-801 is dose- and time-dependent occurring within 2 h and dissipating by 24 h after injection (0.5–8.0 mg/kg, i.p.). The mechanism of this paradoxical induction of c-fos by MK-801 is unclear; however, the pattern of induction appears to follow the distribution of the antagonist-preferring NMDA receptor site.

Induction of c-fos protein-like immunoreactivity in the rat and hamster pineal gland after the onset of darkness

Induction of c-fos protein (FOS) after the onset of darkness was studied immunocytochemically in the rat and hamster pineal gland. The animals were kept on a 12:12 h light-dark cycle. Before the dark period no FOS staining was seen in either rat or hamster pineal cells. Five hours after the onset of darkness 342 +/- 18 pinealocytes/0.2 mm2 (mean +/- SD) displayed FOS-like immunoreactivity in the hamster pineal gland; in the rat pineal gland only 5 +/- 2 pinealocytes/0.2 mm2 showed a faint staining. Two hours later the density of FOS positive cells was decreased to 60 +/- 11/0.2 mm2 in the hamster but increased to 519 +/- 103/0.2 mm2 in the rat pineal gland. Three hours before the beginning of the light period no FOS positive cells were detected in either animal. Both the rat and hamster pineal gland showed a transient and temporally defined expression of c-fos protein in the middle of the dark period. This may be related to a more active functional state of pinealocytes, which is reflected in a peak of melatonin synthesis during the darkness.

D 2 dopamine receptor antagonists induce fos and related proteins in rat striatal neurons

Rats injected with haloperidol, which binds to both D 2 dopamine and sigma receptors or the specific D 2 dopamine receptor antagonist YM 09151-2, but not the specific D 1 dopamine receptor antagonist SCH 23390, showed induction of c-fos protein and c-fos-related antigens in striatal neurons. This effect of haloperidol and YM 09151-2 was inhibited by the N-methyl- d-aspartate antagonist MK801 but was not affected by 1,3-di- O-tolylguanidine, a selective sigma receptor ligand. Two different antisera were used to detect c-fos protein: one was specific for c-fos protein itself while the other recognized c-fos protein as well as c-fos protein-related antigens. In time-course immunocytochemical studies, the c-fos protein was induced maximally by 1 h and had returned to baseline by 24 h. However, c-fos protein-related antigens were induced maximally after 2 h and remained elevated for at least three days after haloperidol injection. Furthermore, the c-fos protein-specific antiserum detected two to three times fewer immunopositive striatal cells than the antiserum which detected both c-fos protein-related antigens and c-fos protein in haloperidol-treated rats. This result suggests that some striatal neurons express c-fos protein-related antigens but not c-fos protein after haloperidol injection. In some striatal sections from haloperidol-injected rats immunostained with the antiserum which recognizes both c-fos protein and c-fos protein-related antigens, there were large areas of immunopositive neurons interspersed with “areas” of striatum devoid of immunostaining. The implications of these results for theories concerning the biochemical mechanism of action of haloperidol are discussed.

Unilateral induction of c-fos protein in cortex following cortical devascularization

The proto-oncogene c-fos is rapidly induced by a variety of stimuli including wounding of the cortex. In an attempt to define the conditions necessary for c-fos activation, we have studied the effects of cerebral devascularization (CD; unilateral pial disruption), a procedure which results in ischaemia without producing direct physical damage to the tissues. Cerebral devascularization produces c-fos activation throughout the ipsilateral cortex; this effect lasts for about 4 h and is not confined to the area subjacent to pial disruption. The activation of c-fos is not 3revented by even high doses of benzodiazepines but is effectively reduced by the N- methyl- d - aspartate (glutamate) receptor antagonist ketamine.

Induction of c-fos protein by activation of vasopressin receptors in smooth muscle cells

Stimulation of vasopressin (V 1) receptors of rat aortic smooth muscle cells (A-10, ATCC CRL 1476) results in the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP 2) to inositol 1,4,5-trisphosphate (IP 3) and diacylglycerol (DAG) with the mobilization of intracellular calcium. When A-10 cells are exposed to arginine vasopressin (AVP), there is an increase in the level of c-fos oncoprotein. The extent of induction of c-fos oncoprotein depends on both the time of exposure of the cells to AVP, reaching a maximum at 60 min after which there is a slow decline, and the concentration of AVP used, with an approximate EC 50 of 1 nM which corresponds well with the K d of vasopressin binding to these receptors. This vasopressin-mediated increase in c-fos protein level is inhibited by a V 1/V 2 antagonist (SKF 101498) suggesting that this is a receptor-mediated event. In addition dDAVP, a V 2 selective agonist, is much less effective than AVP in inducing c-fos protein suggesting that AVP mediates its effect via V 1 receptors. Desensitization of vasopressin receptors by prolonged exposure to AVP resulted in no additional induction of c-fos protein level in response to second challenge of AVP. In addition to AVP, phorbol dibutyrate (PDBu), an activator of protein kinase C (PKC), also stimulates the accumulation of c-fos protein although to a lesser extent than AVP. The above data suggest that c-fos protein levels in smooth muscle cells are regulated by AVP and the hormonal effect may be mediated through PI turnover and DAG, IP 3 and Ca 2+ signals.

High-frequency discharge of dentate granule cells, but not long-term potentiation, induces c-fos protein

‘Competence’ genes, such as c-fos, may play key roles in information storage in the nervous system by linking relatively brief extracellular signals to long-term changes in the neuron. In support of this idea we, and others, have shown that the c-fos protein occurs in adult mammalian neurons and that higher levels of the protein are induced in certain brain regions after kindled or metrazol-induced seizures in mice and rats, sensory stimulation and mechanical damage in spinal cord neurons, and after depolarization in PC12 cells. Here we report that a massive induction of c-fos protein is observed in dentate granule cells in four conditions that result in repetitive firing: localized seizure discharges; high frequency antidromic activation; orthodromic activation in the presence of iontophoresed bicuculline; and frequency potentiation. However, stimulation of the perforant path with high frequency trains that produced long-term potentiation at the perforant path-granule cell synapse did not reliably induce c-fos in the dentate gyrus. These findings suggest that c-fos induction can follow repetitive neuronal discharge but is not involved in long-term potentiation.

Brain injury induces c-fos protein(s) in nerve and glial-like cells in adult mammalian brain

Recent studies have shown that the c-fos protein(s) exists in low basal levels in adult mammalian neurons, but not in glial cells. Here we report that c-fos-protein-like immunoreactivity is induced in glial cells following cortical injury. Glial cells in white matter regions around the wound margin express the c-fos protein maximally at 12–24 h post-insult. Injury is also associated with a massive induction of c-fos proteins in nerve cells in the damaged cerebral cortex. Injections of [ 3H]thymidine into mice and autoradiographic analysis of [ 3H]thymidine incorporation showed that the increase in c-fos preceded glial cell division following injury. These results show that c-fos protein is induced in glial and nerve cells after injury.

Localization and induction of c-fos protein-like immunoreactive material in the nuclei of adult mammalian neurons

The distribution of the proto-oncogene product, c-fos protein, has been studied in tissue from adult rat brain using immunocytochemical techniques. Sections of rat brain were incubated with a polyclonal antibody to a synthetic fragment of the c-fos protein (M-peptide) and visualized for immunoreactivity using the avidin-biotin technique. Low levels of c-fos protein-like immunoreactivity were concentrated in the nucleus. Dark nuclear staining of cells was observed in the cerebral cortex, in hippocampal pyramidal neurons, in granule cells of the dentate gyrus and other regions. Neurons in the cat visual cortex also showed low levels of c-fos protein-like immunoreactivity. Pre-absorbing the antibody with the M-peptide abolished the immunostaining. Generalized seizures evoked by injection of pentylenetetrazol produced a massive induction of fos protein(s) in the piriform and cingulate cortices as well as the dentate gyrus of rats. These results demonstrate that c-fos protein-like immunoreactive materials is found within the nuclei of fully differentiated adult mammalian neurons at low basal levels and that activation of nerve cells leads to an induction of c-fos proteins.

Posttranscriptional regulation of c-fos mRNA expression.

The transient induction of c-fos mRNA and protein suggests that regulation occurs not only by transcriptional activation but also at the level of turnover of the gene product. Here we present evidence for the rapid turnover of c-fos mRNA and some of the requirements for its specific degradation. The half life of induced mature cytoplasmic c-fos mRNA is 9 min in both serum-starved and growing primary human fibroblasts and in NIH 3T3 cells. A structure present at the 3' end of the c-fos mRNA molecule is involved in its low stability since the substitution or the removal of the untranslated 3' portion prolongues the RNA life time. The rapid turnover of fos mRNA requires, in addition, continued protein synthesis. Treatment of cells with cycloheximide stabilizes c-fos mRNA. Washing out cycloheximide reestablishes the rapid turnover. Both changes occur with lag periods of less than 17 minutes.

Microinjection of transforming ras protein induces c-fos expression.

Microinjection of p21ras induced c-fos protein accumulation in three types of 3T3 cells. The induction was rapid and efficient and persisted for many hours. In addition, anti-ras antibody dramatically reduced c-fos accumulation after serum stimulation of injected cells. However, cells which expressed p21ras continuously did not maintain a high level of c-fos expression.

Microinjection of transforming ras protein induces c-fos expression.

Microinjection of p21ras induced c-fos protein accumulation in three types of 3T3 cells. The induction was rapid and efficient and persisted for many hours. In addition, anti-ras antibody dramatically reduced c-fos accumulation after serum stimulation of injected cells. However, cells which expressed p21ras continuously did not maintain a high level of c-fos expression.

Oncogenes, cancer and analytical cytology.

Oncogenes, cancer and analytical cytology.

Induction of c-fos during myelomonocytic differentiation and macrophage proliferation.

Previous studies have suggested a role for c-fos in cellular differentiation in fetal membranes, haematopoietic cells and teratocarcinoma stem cells. In other cell types, such as fibroblasts, c-fos expression is normally very low, but is rapidly induced by peptide growth factors, implicating c-fos in growth control mechanisms. Here, we show that the TPA (12-O-tetradecanoylphorbol-13-acetate)-induced macrophage-like differentiation of HL60 human promyelocytic precursor cells is accompanied by the induction of both c-fos mRNA and protein within 15 min after treatment, suggesting a functional role for c-fos in this differentiation system. In quiescent terminally differentiated macrophages, expression of c-fos is inducible by the macrophage-specific growth factor colony-stimulating factor-1 (CSF-1). The kinetics of c-fos induction, however, are entirely different from those in growth factor-stimulated fibroblasts, supporting the view that the c-fos gene product may serve different functions in different cell types.

Induction of c-fos gene and protein by growth factors precedes activation of c-myc.

Stimulation of fibroblasts with serum or purified growth factors leads to a dramatic induction of expression of both c-fos mRNA and protein within a few minutes, followed by activation of c-myc. This suggests that c-fos induction is a primary event and the earliest known effect on gene expression by growth factors.

Potassium outflux from rabbit cortex during spreading depression.

ArticlesPOTASSIUM OUTFLUX FROM RABBIT CORTEX DURING SPREADING DEPRESSIONF. J. Brinley JR., E. R. Kandel, and Wade H. MarshallF. J. Brinley JR., E. R. Kandel, and Wade H. MarshallPublished Online:01 May 1960https://doi.org/10.1152/jn.1960.23.3.246MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformationCited ByControl of Spreading Depression with Electrical Fields8 June 2018 | Scientific Reports, Vol. 8, No. 1Inadequate brain glycogen or sleep increases spreading depression susceptibility24 January 2018 | Annals of Neurology, Vol. 83, No. 1The CGRP receptor antagonist BIBN4096 inhibits prolonged meningeal afferent activation evoked by brief local K+ stimulation but not cortical spreading depression-induced afferent sensitizationPAIN ReportsSpreading Depression, Spreading Depolarizations, and the Cerebral VasculatureCenk Ayata and Martin Lauritzen1 July 2015 | Physiological Reviews, Vol. 95, No. 3Mechanisms of Spreading Depression and Hypoxic Spreading Depression-Like DepolarizationGeorge G. Somjen1 July 2001 | Physiological Reviews, Vol. 81, No. 3Unilateral induction of c-fos protein in cortex following cortical devascularizationBrain Research, Vol. 503, No. 2Extracellular K+ accumulation in the central nervous systemProgress in Biophysics and Molecular Biology, Vol. 42Functional interactions between neurons and astrocytes. II. Potassium homeostasis at the cellular levelProgress in Neurobiology, Vol. 20, No. 1-2Functional interactions between neurons and astrocytes I. Turnover and metabolism of putative amino acid transmittersProgress in Neurobiology, Vol. 13, No. 3Extracellular ionic variations during spreading depressionNeuroscience, Vol. 3, No. 11Potassium activity in rabbit cortexBrain Research, Vol. 75, No. 1Reactivity of penicillin-induced epileptogenic foci to selective blockade of membrane sodium-potassium ATPase via ouabainExperimental Neurology, Vol. 42, No. 3A comparative study of membrane potential changes in neurons and neuroglial cells during spreading depression in the rabbitBrain Research, Vol. 65, No. 3Measurement of extracellular potassium activity in cat cortexBrain Research, Vol. 50, No. 2Potassium-selective microelectrodes used for measuring the extracellular brain potassium during spreading depression and anoxic depolarization in ratsBrain Research, Vol. 39, No. 1Potassium accumulation in interstitial space during epileptiform seizuresExperimental Neurology, Vol. 26, No. 3Spreading depression in the striatum and optic lobes of the chickenBrain Research, Vol. 13, No. 1Hippocampal epileptic activity induced by localized ventricular perfusion with high-potassium cerebrospinal fluidExperimental Neurology, Vol. 20, No. 1Blockade of thalamocortical and pyramidal pathways by striatal spreading depression in ratsExperimental Neurology, Vol. 18, No. 4Spreading depression in subcortical structures in rabbitBrain Research, Vol. 2, No. 1Intracellular potentials from “idle” cells in cerebral cortex of catElectroencephalography and Clinical Neurophysiology, Vol. 20, No. 6Thalamic spreading depression in the ratElectroencephalography and Clinical Neurophysiology, Vol. 20, No. 1Reversible deafferentation of cerebral cortex by thalamic spreading depressionExperimental Neurology, Vol. 12, No. 1Specific impedance of cerebral cortex during spreading depression, and an analysis of neuronal, neuroglial, and interstitial contributionsExperimental Neurology, Vol. 9, No. 1Influence of transcortical d-c currents on cortical neuronal activityExperimental Neurology, Vol. 5, No. 6Inhibitory action of carbon dioxide and acetazoleamide in seizure activityElectroencephalography and Clinical Neurophysiology, Vol. 13, No. 5 More from this issue > Volume 23Issue 3May 1960Pages 246-256 https://doi.org/10.1152/jn.1960.23.3.246PubMed13804482History Published online 1 May 1960 Published in print 1 May 1960 Metrics