[Show abstract][Hide abstract] ABSTRACT: Fibroblast growth factor-2 (FGF-2) is predominantly synthesized and secreted by astrocytes in adult brain. Our previous study showed that activation of classical dopamine receptor D(1) or D(2) elicits FGF-2 biosynthesis and secretion in astrocytes. Here, we report that astrocytic FGF-2 expression is also regulated by phosphatidylinositol (PI)-linked D(1)-like receptor. SKF83959, a selective PI-linked D(1)-like receptor agonist, upregulates the levels of FGF-2 protein in striatal astrocyte cultures in classical dopamine D(1) and D(2) receptor-independent manner. The conditional medium derived from SKF83959-activated astrocytes promoted the number of TH(+) neurons in vitro. Treatment of astrocytes with SKF83959 increased intracellular calcium in two phases. Inhibition of intracellular calcium oscillation by inositol 1,4,5-triphosphate (IP3) inhibitors blocked the SKF83959-induced increase in FGF-2 expression. Moreover, intraperitoneal administration of SKF83959 reversed l-methyl-4-phenyl-l,2,3,6-tetrahydropypridine (MPTP)-induced reduction in FGF-2 expression in both the striatum and ventral midbrain and resulted in marked protection of dopaminergic neurons from MPTP-induced neurotoxicity. These results indicate that IP3/Ca(2+)/calmodulin-dependent protein kinase is an uncharted intracellular signaling pathway that is crucial for the regulation of FGF-2 synthesis in astrocytes. PI-linked D(1)-like receptor plays an important role in the regulation of astrocytic FGF-2 expression and neuroprotection which may provide a potential target for the drug discovery in Parkinson's disease.
Journal of Neuroscience 07/2009; 29(24):7766-75. · 6.91 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Apomorphine (APO) is an anti-parkinsonian drug currently in use, which provides relief of Parkinson's symptoms. However, the utility of APO is greatly hampered by its poor bioavailability and rapid metabolism. In the present study, O,O'-diacetyl-apomorphine, a prodrug of apomorphine, was synthesized and its biological activity was examined. The prodrug induced fibroblast growth factor-2 production in astrocytic cultures similarly to apomorphine. However, its duration of action was significantly prolonged, and its resistance to oxidation was markedly enhanced compared to APO. O,O'-Diacetyl-apomorphine also induced MEK/MAPK signaling. These results suggest that O,O'-diacetyl-apomorphine can efficiently counteract oxidation and thereby enhance FGF-2 production in astrocytes.
Biochemical and Biophysical Research Communications 06/2008; 369(3):824-9. · 2.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Apomorphine (APO), a potent D1/D2 dopamine receptor agonist, is currently used as an antiparkinsonian drug. We have shown previously that APO stimulates synthesis and release of multiple trophic factors, such as brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF), in both mesencephalic and striatal neurons, thereby effectively preventing dopaminergic neuron loss in vitro. The present study was designed to investigate the effects of APO on fibroblast growth factor-2 (FGF-2) expression and regulation in astrocytes, and furthermore, to identify signaling mechanisms underlying these effects. Here, we show that FGF-2 expression is robustly induced in cultured astrocytes in response to APO. FGF-2 expression was proportional to APO concentration and time-dependent. Conversely, treatment with S-APO, a derivative of R-APO lacking DA receptor agonist activity, did not alter FGF-2 levels. APO treatment resulted in enhanced cytosol FGF-2 immunoreactivity, export of high MW forms of FGF-2 to the cytoplasm from the nucleus and increased extracellular release of FGF-2. Interestingly, both high and low MW forms of FGF-2 were detectable in conditioned medium of APO-treated cultures. This APO-induced effect was correlated with activation of D1 and D2 receptors, as it could be either mimicked by dopamine receptor agonists (SKF38393, quinpirole) or partially blocked by antagonists (SCH23390, SKF83566, haloperidol). Activation of the D1 receptor preferentially increased PKA activity, whereas activation of the D2 receptor only promoted phosphorylation of MAPK. Importantly, APO-modulated FGF-2 expression was independent of Akt/phosphoinositide 3-kinase signaling. These data suggest that APO can enhance biosynthesis and release of FGF-2 through activation of dopamine receptors in striatal astrocytes. Both cAMP/PKA and MEK/MAPK signaling cascades are major steps mediating this process.
The FASEB Journal 07/2006; 20(8):1263-5. · 5.70 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Bystin has been identified as a protein which mediates cellular interactions between trophoblastic and endometrial epithelial cells by forming complexes with two partners, trophinin and tastin, during embryo implantation. However, the presence of bystin in the central nervous system has not been demonstrated. Here, we report the cloning of the full-length cDNA of the rat bystin gene from adult brain. Immunohistochemical and RT-PCR analysis showed that the levels of bystin expression were markedly up-regulated in the both 6-hydrodopamine-lesioned rat nigrostriatum and stab-lesioned cerebral cortex in adult rats. Double immunofluorescence staining revealed that most bystin-expressing glial cells were astrocytes (immature or mature). To determine the mechanisms for the up-regulation of bystin expression in glial cells, primary cultures of postnatal cortical astrocytes were employed. Western blot analysis showed that the expression of bystin was elevated by treatment with pro-inflammatory mediators lipopolysaccharide and interleukin-1 beta. Nerve growth factor known to be released after brain injury also induced bystin expression in the cultures. Exposure of astrocyte cultures to the differentiating agent forskolin resulted in up-regulation of bystin followed by a pronounced astrocytic stellation. The results suggest that the injury in the adult brain induces spatiotemporal up-regulation of bystin and it could be influenced, at least in part, by elevation of intracellular cAMP level. Bystin expressed by reactive astrocytes may be involved in their differentiation during the inflammatory processes following brain injury. The reappearance of bystin may also indicate that some reactive astrocytes have the capacity to recapitulate early developmental stages.
European Journal of Neuroscience 09/2004; 20(4):873-84. · 3.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated
with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubule-associated
proteins (MAPs) and functionally link to microtubule, we first successfully identified JWA from the physically purified MAPs
complex of rat brain tissues. The results of co-immunoprecipitation, gene transfection and immunofluorescence microscopy assays
from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and β-tubulin. In general, JWA is stably binding
to β-tubulin whenever microtubule is polymerized or not, and it may be critical to the mitosis process. In addition, by use
of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellular amino acids
in PC 12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory amino acid, and glutamate,
an excitatory amino acid. In conclusion, JWA is not only structurally associated, but also a novel functional MAP.
KeywordsJWA-β-tubulin-microtubule-associated proteins (MAPs)-amino acid transportation
Chinese Science Bulletin 09/2003; 48(17):1828-1834. · 1.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Our previous study elucidated that JWA protein was a newly identified microtubule-associated protein (MAP), which combined
to and co-localized with β-tubulin. In the present study, we designed a series of experiments to explore if any interactions
between JWA protein and α-tubulin existed and how JWA protein would functionally link to α-tubulin, especially in cell mitosis.
Results of coimmunoprecipitation, gene transfection and immunofluorescence microscopy from PC12 and HEK293 cells provided
strong evidence for a linkage between JWA protein and α-tubulin. Our data showed that JWA protein bound to α-tubulin stably
no matter whether α-tubulin was polymerized or not. In addition, by using antisense oligonucleotides, cell cycle blocking
agents and hypothermia disposal techniques, we also found the interaction between JWA protein and α-tubulin. The further analysis
using flow cytometry and confocal microscopy showed that both proteins co-existed in PC12 cells and were independent on the
cell cycle. In conclusion, JWA protein is a newly identified microtubuleassociated protein, binds to α-tubulin, and probably
plays an important role in regulation of microtubular stability.
Chinese Science Bulletin 49(5):467-471. · 1.37 Impact Factor