Purpose: No ideal method for subdividing and assessing changes in neurons of the spinal cord during specific conditions has been established. We attempted to develop a method for subdividing spinal neurons using immunohistochemical and fluorescent staining, which is an important key towards understanding the mechanism of reflex voiding. Materials and Methods: Thirty Sprague-Dawley rats, weighting 200-300g, were divided into five groups. A cystometrogram was performed during saline or acetic acid instillation. We identified the neuronal pathway associated with the detrusor by injecting a pseudorabies virus (PRV) into the detrusor muscle and inspecting the changes in relation to different time sequences. An immunohistochemical staining method was used to stain the fos-protein encoded by the c-fos gene. Immunofluorescent staining was performed to evaluate changes in the neurons in relation to the voiding reflex, and the neurons then subdivided. Results: We confirmed pseudorabies virus (PRV) infection of the cells in the sacral parasympathetic nucleus through immunohistochemical staining two days after injection. On detection of an increase in c-fos positive cells after dividing the c-fos positive area of the L6 and S1 spinal cord into 4 sections, significant increases were observed in the sacral parasympathetic nucleus (SPN) and dorsal commissure (DCM). Double staining was performed to detect the neurons associated with the voiding reflex in the SPN and DCM areas showing overexpression of c-fos. Conclusions: The establishment of a method for detecting morphological changes, and subdividing neurons by immunohistochemical and fluorescent staining, may provide an important key towards understanding the mechanism of various neuromodulations of clinically applied treatments. (Korean J Urol 2005;46:487-494)
Acetic Acid ; Genes, fos ; Herpesvirus 1, Suid ; Neurons* ; Proto-Oncogene Proteins c-fos ; Rats, Sprague-Dawley ; Reflex ; Spinal Cord

Some proto-oncogenes in human malignancies can be primary cause of carcinogenesis and can be correlated to histologic aggressiveness or prognosis. In this study, to define the role of c-myc and c-fos protooncogenes in multistage tumorigenesis of thyroid tumor and to correlate the expression of c-myc protooncogene with the degree of histologic aggressiveness and EORTC prognostic index in patients of thyroid carcinoma, 132 fresh specimens obtained at surgery from 71 cases of thyroid tumor were analysed by slot blot hybridization for the expression of c-myc and c-fos proto-oncogene, and 42 formalin-fixed paraffin-embedded tissues were analysed by immunohistochemical staining with monoclonal c-myc antibody and monoclonal Ki-67 antibody, and in 34 paraffin-embedded malignant tissues, the degree of histologic aggressiveness was determined, and in 42 cases of thyroid cancer, EORTC prognostic index were determined. And each item was compared to c-myc mRNA expression level. The results obtained were as follows: 1. The level of c-myc mRNA was significantly higher in malignant thyroid tissues as compared to normal or benign thyroid tissues(p=0.01). 2. The level of c-fos mRNA was similar in normal, benign and well differentiated thyroid carcinoma 3. There was no difference of c-myc mRNA expression levels between central and peripheral tumor tissues. 4. C-myc expression of normal tissues originated from cancer was higher than those from benign tumor(p=0.01) 5. A high level of c-myc mRNA in differentiated thyroid cancer corresponded to a high degree of histologic aggressiveness, although there was no statistically significant correlation between two. 6. High levels of c-myc mRNA were more frequently found in thyroid carcinoma with unfavourable prognosis, although there was no statistically significant correlation between c-myc mRNA expression and EORTC prognostic index or other prognostic factors such as age, sex, tumor size, tumor extent in thyroid carcinoma. 7. C-myc mRNA expression was statistically significantly correlated to the degree of immunohistochemical staining of monoclonal c-myc antibody(p=0.01). 8. C-myc mRNA expression was statistically significantly correlated to Ki-67 cell proliferative index score(p=0.005). On the basis of results, it can be suggested that overexpression of c-myc protooncogene in thyroid carcinoma is secondary phenotype of growth signal rather than primary cause of thyroid carcinogenesis, and expression of c-fos protooncogene was similar in normal, benign and malignant tumor tissues. C-myc may be useful prognostic indicator because there was some relationship between c-myc mRNA expression and EORTC prognostic index score or histologic aggressiveness score. And because c-myc correaltes significantly to Ki-67 cell proliferative index score, c-myc expression will be important growth signal in follicular cell of thyroid and have a role of progression of thyroid carcinoma.
Carcinogenesis ; Genes, fos ; Humans* ; Phenotype ; Prognosis ; Proto-Oncogenes ; RNA, Messenger ; Thyroid Gland* ; Thyroid Neoplasms

OBJECTIVETo explore the mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myocardial ischemia.

METHODSThe rats were randomly divided into a sham operation group, a myocardial ischemia model group and a myocardial ischemia model plus electroacupuncture group. The acute myocardial ischemia model was developed byligation of the descending anterior branch of the coronary artery, and electroacupuncture was given at bilateral "Neiguan" (PC 6). Serum myocardial enzymes was determined by biochemical method and the expression of c-fos mRNA in myocardium was detected by using reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe activities of serum myocardial enzymes and the expression of c-fos mRNA in ischemic myocardium were significantly increased as compared with those in the sham operation group (P < 0.05), and after electroacupuncture they were significantly decreased (P < 0.05).

CONCLUSIONThe mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myochadial ischemia is possibly related with down-regulation of expression of c-fos mRNA in myocardium.

Acupuncture Points ; Animals ; Electroacupuncture ; Genes, fos ; Humans ; Myocardial Ischemia ; genetics ; Myocardium ; metabolism ; Rats

OBJECTIVETo investigate the relationship between c-fos gene and filamentous actin (F-actin) in MG-63 osteoblasts under cyclic tensile stress.

METHODSMG-63 osteoblasts were subjected to cyclic tensile stress (0.5 Hz, 2 000 microstrain) for 3, 6, and 12 h. The changes of c-fos gene were investigated by fluorescent quantitation polymerase chain reaction. Then the best loading time group was screened as the experimental group compared with 0 h group. The changes of F-actin and c-fos were investigated with or without cytochalasin D treatment.

RESULTSCyclic tensile stress induced high expression of c-fos mRNA, and peaked at 3 h. After loading, F-actin had a structure reorganization, but had no change in expression. After cytochalasin D treatment, the formation of stress fibers and the fluorescence intensity of F-actin cytoskeleton significantly reduced, meanwhile the c-fos mRNA expression was inhibited.

CONCLUSIONAfter loading, there is only structure reorganization for F-actin, and the expression has not any change. That means the remodeling F-actin is the existing one. F-actin reorganization is an important part in c-fos gene expression induced by stress.

Actin Cytoskeleton ; Actins ; Cytochalasin D ; Cytoskeleton ; Genes, fos ; Humans ; Microtubules ; Osteoblasts ; RNA, Messenger ; Stress, Mechanical

BACKGROUND AND OBJECTIVES: Fos, the protein product of c-fos gene, has been known to be rapidly expressed in neurons following various external and internal stimuli and this protein expression has been used as a neural activation marker in many fields. This experiment was performed to examine the afferent pathway of the lateral semicircular canals following sinusoidal acceleration. MATERIALS & METHODS: To stimulate the lateral semicircular canals, animals received rotary stimulation for 90 minutes with 2.0Hz sinusoidal acceleration. Thirty minutes after stimulation, the subjects were sacrificed and their brainstems were processed for immunohistochemistry to detect Fos expression. RESULTS: Fos proteins were strongly expressed in the superior, dorsal medial vestibular and inferior vestibular nuclei. However, there was no expression in the lateral and ventral portion of medial vestibular nuclei. CONCLUSION: This finding suggested that the afferent pathway from peripheral vestibular end-organ can be successfully mapped by detection of Fos expression and Fos is an useful neural activity marker in the vestibular system.
Acceleration* ; Afferent Pathways ; Animals ; Brain Stem* ; Genes, fos ; Immunohistochemistry ; Neurons ; Semicircular Canals ; Vestibular Nuclei*

Fos, the protein product of c-fos gene, has been known to be rapidly expressed in neurons following various external and internal stimuli. This experiment was performed to determine whether off-axis centripetal acceleration cause a proper stimulation to the otolith organ and the afferent neural pathway from the otolith organ can be mapped by detection of expressed Fos protein. Therefore we performed on-axis and off-axis centripetal acceleration stimulation to experimental animals and investigated Fos expression immunohistochemically. In results, there were no expression of Fos protein following on-axis centripetal acceleration, whereas in the medial, descending vestibular nuclei and y-group of the off-axially stimulated animal, we could find strong expression of Fos protein.
Acceleration* ; Animals ; Brain Stem* ; Genes, fos ; Neural Pathways ; Neurons ; Otolithic Membrane ; Rats* ; Vestibular Nuclei*

AIMTo investigate the effects of anoxia/reoxygenation on Fos and Jun expression and apoptosis in cultured rat hippocampal neurons.

METHODSThe hippocampal neurons cultured for 12 d were exposed to anoxia environment (90% N2 + 10% CO2) for 4 h and then reoxygenated for 24 h and 72 h. The neurons were immunocytochemically stained using the antiserum against Fos and Jun, and the apoptosis were detected by using the terminal deoxynucleotidyl transferase mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL) method and flow cytometric analysis.

RESULTSThe percentage of Fos and Jun positive neurons and apoptosis neurons in cultured hippocampal neurons after anoxia/reoxygenation increased than those in control.

CONCLUSIONThe occurrence of neurons apoptosis is related to the increase in Fos and Jun expression in cultured hippocampal neurons after anoxia/reoxygenation.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Genes, fos ; Genes, jun ; Hippocampus ; metabolism ; Neurons ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of transforming growth factor beta (TGF ) on c-fos gene expression in hepatic stellate cells.

METHODSHepatic stellate cells (HSC-T6) were cultured in the medium containing different concentrations of TGF (0.2, 1, and 5 ng/ml), and cells were collected at different time points of incubation (8, 24, 48, and 72 h). The total RNA of the HSCs was isolated and c-fos gene expression level were measured by reverse transcription polymerase chain reaction.

RESULTSc-fos gene expression levels of HSCs cultured in the presence of low (0.2 ng/ml), moderate (1 ng/ml) and high (5 ng/ml) concentrations of TGF for 8, 24, 48 and 72 h were significantly greater than those of control group. The c-fos gene expression levels of HSCs increased gradually with the increment of TGF concentration, and significant differences in c-fos gene expression were found between the 3TGF groups.

CONCLUSIONTGF strongly up-regulates c-fos gene expression in hepatic stellate cells.

Animals ; Cells, Cultured ; Gene Expression ; drug effects ; Genes, fos ; Hepatic Stellate Cells ; drug effects ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Rats ; Transforming Growth Factor beta ; pharmacology

The epidemiology of cancer has long been suggested that cancer is multistep disease. We suspect some of these steps might be lated with activation of oncogenes and loss of tumor suppressor genes in primary brain tumors. Moreover, recent reports suggest that astrocytomas have shown alterations in chromosome 17p, and this chromosomal location that encodes the p53 protein, as well as c-fos gene may take an important role in the carcinogenesis of human primary brain tumors. Expression of p53 protein was detected in 12 of 17 cases(70.6%) of glioblastoma multiforme, 4 of 6 cases(66.6%) of anaplastic astrocytoma with positive nuclear p53 staining. All low grade astrocytomas and normal brain tissue failed to express p53. Correlation of p53 protein levels with mRNA alterations or genomic DNA alterations may help to guide future therapy or diagnosis of brain tumors. On the other hand, the level of c-fos oncoprotein expression may be correlated with the degree of cell differentiation and proliferation. The presence of these expression in low-grade astrocytoma suggest that activation of the c-fos gene is an early step in tumor development.
Astrocytoma ; Brain Neoplasms ; Brain* ; Carcinogenesis ; Cell Differentiation ; Diagnosis ; DNA ; Epidemiology ; Genes, fos ; Genes, Tumor Suppressor ; Glioblastoma ; Glioma* ; Hand ; Humans ; Immunohistochemistry ; Oncogenes ; RNA, Messenger

OBJECTIVETo investigate the expression sequence and distribution characteristics of the protooncogenes c-fos, c-myc and endogenous basic fibroblast growth factor (bFGF) genes in burned tissues, and to explore the possible effects of changes in these genes' functions on wound healing.

METHODSPartial-thickness burns of 30% TBSA were established on backs of Wistar rats. In situ hybridization and histological methods were used to detect expression of c-fos, c-myc and bFGF genes in normal and burned tissue at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d postburn.

RESULTSAlthough expression of c-fos and c-myc genes and bFGF gene could be found in normal skin, the expression of all three were markedly induced by burn wounds and the expression models in sequence and distribution were quite different. Expression of c-fos gene increased and peaked at 6 h. Signals were mainly localized in both nuclei of dermal fibroblasts and monocytes. The expression of bFGF gene increased at 6 h and peaked at 1 d postburn, and was distributed in the cytoplasm of fibroblasts. C-myc gene peaked 3 d postburn and was also distributed in the cytoplasm of fibroblasts.

CONCLUSIONSThese results indicated that thermal injury could induce the expression of c-fos, c-myc and bFGF at gene level, showing phasic control and regional distribution. The phasic expression of these genes suggests that there is an interaction between protooncogenes and bFGF, which may play an important role in wound healing. The different expressions of c-fos and c-myc play an inducing role in regulating bFGF, and in turn affect wound healing.

Animals ; Burns ; metabolism ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation ; Genes, fos ; Genes, myc ; In Situ Hybridization ; Male ; Rats ; Rats, Wistar ; Time Factors