1.Expression of c-fos and c-jun proteins in the marginal division of the rat striatum during learning and memory training.
Xin-min BAO ; Si-yun SHU ; Hong WANG
Chinese Medical Journal 2005;118(5):398-403
BACKGROUNDA new brain region, the marginal division (MrD), was discovered at the caudal margin of the neostriatum. The MrD was shown to be involved in learning and memory in the rat. The aim of this study was to investigate the expression of the immediate-early genes c-fos and c-jun in the MrD of the striatum during learning and memory processes in the rat, immunocytochemical and Western blot methods were used to examine Y-maze trained rats.
METHODSThe rats were divided into three groups, namely the training, pseudotraining, and control groups. After Y-maze training, the expression of the immediate-early genes c-fos and c-jun in the MrD of the rats was investigated using immunocytochemical and Western blot methods.
RESULTSAfter one hour of Y-maze training, the expression of c-jun and c-fos proteins was significantly enhanced in the MrD; the c-jun protein, in particular, was more intensely expressed in this region than in other parts of the striatum. The expression of these two proteins in the training group was significantly higher than in the pseudotraining and control groups. In addition, positive expression was also found in the hippocampus, cingulum cortex, thalamus, and in other areas. Western blot disclosed two immunoreactive bands for the anti-c-fos antibody (47 kD and 54 kD) and two immunoreactive bands for the anti-c-jun antibody (39 kD and 54 kD).
CONCLUSIONSThese results indicate that the immediate-early genes c-fos and c-jun participate in signal transduction during the learning and memory processes associated with Y-maze training in rats.
Animals ; Male ; Maze Learning ; Memory ; Neostriatum ; metabolism ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis ; Rats ; Rats, Sprague-Dawley
2.Preparation and application of goat deltafosB gene expression product antibody.
Huiling ZHENG ; Zhenzhen ZHU ; Junhui AN ; Zhenyu YANG ; Ruifang XING ; Linhui YAN
Chinese Journal of Biotechnology 2010;26(12):1704-1709
deltaFosB, a naturally occurring truncated isform of fosB gene, existed in many tissues stably and played an important role in formation and differentiation of adipocyte and osteoblast. deltaFosB may be related to the metabolism of calcium in bone and mammary gland and regulate the signal pathway of calcium transfer from bone to mammary gland. We first sub-cloned deltafosB gene of goat into the vector pET32a to construct prokaryotic expression vector pET32a-deltafosB. Then we induced for deltafosB gene expression efficiently by IPTG. Finally we immunized the adult rabbits with purified recombinant deltaFosB to prepare rabbit anti-goat deltaFosB polyclonal antibody. iELISA analysis showed the antibody with the titer of 1:51 200, and Western blotting result showed that the antibody could specifically detect the deltaFosB protein expressed in prokaryotic cell and HEK-293 cell, respectively. Further Western blotting assay showed that deltaFosB expressed in various tissues of goat in vivo.
Animals
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Antibodies, Monoclonal
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biosynthesis
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Gene Expression
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Goats
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Proto-Oncogene Proteins c-fos
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biosynthesis
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genetics
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immunology
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Rabbits
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
4.Expression of c-fos, OPG, OPGL in rabbit mandibular distraction osteogenesis zone.
Wei-li GE ; Zhi-jian XIE ; Jian-feng HE
Journal of Zhejiang University. Medical sciences 2006;35(5):496-500
OBJECTIVETo evaluate the possible signal transduction mechanism of the mechanical stress induced by the distraction procedure in osteocytes.
METHODSAn animal model of mandibular distraction osteogenesis in rabbits was established. The expressions of c-fos, OPG and OPGL were detected by ultrasensitive S-P immunohistochemical method.
RESULTAt 4 and 8 days after distraction, distraction zone showed strong positive staining of c-fos, which were apparently higher than that in distraction zone of 2, 4 and 6 weeks after consolidation. At 4 and 8 days after distraction and 2 weeks after consolidation, the expression of OPG was strong, and then wore off gradually at 4 and 6 weeks after consolidation. Weak signals of OPGL could be detected at 6 weeks after consolidation only.
CONCLUSIONc-fos, OPG and OPGL are important regulators in distraction osteogenesis. c-fos is interrelated with the mechanical stress induced by the distraction procedure closely, OPG promotes new bone formation, while OPGL plays a more active role in bone remodeling.
Animals ; Mandible ; cytology ; metabolism ; Osteocytes ; metabolism ; Osteogenesis, Distraction ; Osteoprotegerin ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RANK Ligand ; biosynthesis ; genetics ; Rabbits ; Random Allocation
5.The mechanotransduction mechanism of how osteoblasts respond to mechanical stimulation.
Yuanliang WANG ; Liling TANG ; Jianhua WANG ; Shaoxi CAI
Journal of Biomedical Engineering 2005;22(2):400-402
The stress environment regulates the factors of growth, resorption and remolding in bone tissue. Mechanical stimulation at cell physical level affects the physiological activity of osteoblasts, including proliferation, ALP activity and osteocalcin production. Mechanotransduction is a procedure which transduces the biophysical force into biochemical responses. It is also the basis of many physiological functions. The early response genes (c-fos, c-jun), the second message systems (Ca2+, NO, cAMP) and the mechano-sensitive cation channel are involved in the mechanotransduction course when osteoblasts respond to the mechanical stimulation.
Biomechanical Phenomena
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Calcium
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physiology
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Humans
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Mechanotransduction, Cellular
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Osteoblasts
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physiology
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Osteocalcin
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biosynthesis
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Proto-Oncogene Proteins c-fos
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biosynthesis
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genetics
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Proto-Oncogene Proteins c-jun
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biosynthesis
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genetics
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Signal Transduction
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Stress, Mechanical
6.Effects of microcystins on cell cycle and expressions of c-fos and c-jun.
Chinese Journal of Preventive Medicine 2003;37(1):23-25
OBJECTIVESTo investigate the effects of microcystins on cell cycle and expressions of c-fos and c-jun, and explore the potential carcinogenic mechanisms of Microcystins.
METHODSMicrocystic cyanobacteria extraction (MCE) purified by Sep-Pak C(18) cartridge was added into the media and co-incubated with SHE cell for various periods. Immunohistochemistry assay was applied to detect the expressions of c-fos and c-jun at 1, 3, 6 hr time point, and cell cycle at 6, 12, 24 hr point were analyzed by flow cytometry respectively.
RESULTSSustained up-regulated expression of c-fos and c-jun were induced by MCE during the experimental period, and 5 - 6 folds increased expression were observed at 6 hour point after treatment. As much as 44.8 per cent of cells were induced to entry S-phase from resting G(0)/G(1).
CONCLUSIONUp-regulating the expression of transcript factor such as c-fos and c-jun thus to induce cell abnormal proliferation may be the potential carcinogenic mechanisms of microcystins.
Animals ; Carcinogens ; toxicity ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Cricetinae ; Gene Expression ; drug effects ; Microcystins ; Peptides, Cyclic ; toxicity ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis
7.DNA damage and oncogenic protein expression induced by cadmium.
Feng CAO ; Tong ZHOU ; Tai-yi JIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(10):587-590
OBJECTIVETo explore the DNA damage and the expression of oncogenic protein induced by cadmium in vitro (human cells) and in vivo in rats.
METHODSThe colony formation assay and the MTT assay were employed to determine the cytotoxicity of cadmium. The DNA damage and the cell cycle were measured by the comet assay and the flow cytometry, respectively. The western bolt and the X-Gal staining were also used to determine the change of oncogenic protein and the senescent marker.
RESULTSThe cadmium inhibited the proliferation of cells and induced DNA damage significantly not only in human cultured cells but also in vivo animal cells. The comet rate increased from 6.1% in the control group to 23.2% in 200 microM cadmium treatment group (P < 0.01). The comet rate increased in all organs of male rats with the increase of dosage, and there were significant difference between treatment the groups and the control group in kidney and ventral prostate (P < 0.05). Furthermore, the cadmium blocked the cell cycle progression. At the same time, the expression of c-myc, c-Jun and beta-Gal were increased by cadmium.
CONCLUSIONThe cadmium could induce DNA damage and block the cell cycle, and further cause senescence, and death. However, the cadmium induced the DNA damage and the oncogenic protein expression may be the important factors to cause cancer.
Animals ; Cadmium ; toxicity ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Comet Assay ; DNA ; drug effects ; DNA Damage ; Dose-Response Relationship, Drug ; Male ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis ; Rats ; Rats, Wistar
9.The effects of genistein on epidermal growth factor receptor mediated signal transduction pathway in human ovarian carcinoma cells lines SKOV3 and its xenograft in nude mice.
Yu LI ; Can MI ; Yong-Zhong WU ; Su-Fen YANG ; Zheng-Qin YANG
Chinese Journal of Pathology 2004;33(6):546-549
OBJECTIVEIt was reported previously that genistein could inhibit proliferation of human ovarian carcinoma cell line SKOV(3), but mechanism was not clear. There is a close relationship between EGFR mediated signal transduction pathway and the development of ovarian tumor. This study aimed to investigate the effects of genistein on the EGFR mediated signal transduction pathway and to clarify its mechanisms of proliferation inhibition on human ovarian carcinoma cell line SKOV(3) and its xenograft in nude mice.
METHODSThe expression of c-erbB-2 protein was determined using immunocytochemistry. The expressions of c-jun and c-fos protein were determined using Western blotting. The expression of c-erbB-2, c-raf-1, c-jun and c-fos mRNA were tested by reverse transcription-polymerase chain reaction (RT-PCR).
RESULTSThe expression of c-erbB-2, c-raf-1 and its downstream gene c-jun and c-fos were decreased at mRNA level in the 20 micromol/L genistein group. The expression of c-erbB-2 protein were decreased, its average absorbency (A) were decreased after treatment of SKOV(3) with 20 micromol/L genistein for 48 h, reached at 0.42 +/- 0.02 (P < 0.05). Western blotting demonstrated that the expressions of c-jun and c-fos protein were decreased gradually after being treated with 20 micromol/L genistein for 12 - 72 h.
CONCLUSIONSGenistein could down-regulate the expression of two key genes, c-erbB-2 and c-raf-1 at mRNA and protein level in the EGFR mediated signal transduction pathway, and down-regulate the expression of its downstream nuclear transcription factors c-jun and c-fos at mRNA and protein level. It is suggested that interfering the expressions of some key signal molecules in EGFR mediated signal transduction system by genistein may account for its molecular foundation of proliferation inhibition in ovarian carcinoma.
Animals ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Genes, fos ; Genes, jun ; Genistein ; pharmacology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Ovarian Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-raf ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Signal Transduction ; drug effects
10.Influence of stretch strain and pressure on expression of osteoblasts' FOS protein.
Xizheng ZHANG ; Yong GUO ; Reixin LI ; Yunshan ZHAO ; Jimin WU ; Jing GUAN
Journal of Biomedical Engineering 2006;23(2):326-328
Uniaxial stretch strain and compressive pressure of 2 atm were applied to rat's osteoblasts, and then immunohistochemistrical staining was used to detect the expression of osteoblasts' c-fos gene. The experiment result indicated the osteoblasts' FOS proteins increased prominently, and the FOS proteins concentrated in nucleolus after having endured two different kinds of loadings. It is very important to prompt stress and strain in promoting osteoblast proliferation.
Animals
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Animals, Newborn
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Cell Proliferation
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Cells, Cultured
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Osteoblasts
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cytology
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metabolism
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Proto-Oncogene Proteins c-fos
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biosynthesis
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genetics
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Rats
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Rats, Wistar
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Skull
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cytology
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Tensile Strength