Differences in the regulation of BDNF and NGF synthesis in cultured neonatal rat astrocytes.
Using a new brain-derived neurotrophic factor (BDNF) specific enzyme-immunoassay, we determined the basal cellular content of BDNF protein in neonatal rat astrocytes in primary culture, thus confirming the ability of astrocytes to synthesize BDNF in addition to nerve growth factor (NGF). We subsequently monitored the influence of different pharmacological agents: neurotransmitter receptor agonists, cytokines, and second messenger up-regulators, on the synthesis of BDNF and NGF. Marked differences in the regulation of their synthesis by the above pharmacological agents were observed in our study. The basal cellular levels of BDNF protein in cultured neonatal rat cortical and cerebellar astrocytes were 15.9 +/- 0.3 and 18.7 +/- 0.4 pg BDNF/mg cell protein, respectively, and differ significantly between astrocytes from different brain regions, whereas NGF levels were the same (16.1 +/- 0.3 and 16.2 +/- 0.7 pg NGF/mg cell protein, respectively). Screening different neurotransmitter systems for their influence on BDNF and NGF synthesis in cortical astrocytes revealed that dopamine (0.15 mM) is a potent up-regulator of BDNF protein synthesis in astrocytes, while kainic acid (50 microM) and histamine (1 microM) did not raise the cellular level of BDNF protein. Dopamine had no influence on NGF synthesis, while kainic acid caused minor, and histamine marked, elevation of NGF cellular content. Tumor necrosis factor-alpha (30 ng/ml) and interleukin-1beta (10 U/ml) treatments did not influence BDNF synthesis, whereas they markedly increased NGF protein cellular level. We also confirmed (using forskolin (20 microM) and phorbol 12-myristate 13-acetate (TPA) (100 nM)) that adenylate cyclase and protein kinase C participate in the downstream signaling responsible for the stimulation of BDNF synthesis, whereas in the regulation of NGF synthesis only the participation of protein kinase C was confirmed. Our results indicate that astrocyte-derived neurotrophins could play a role in distinct brain functions under physiological conditions and in the pathogenesis as well as possible treatment of different neurodegenerative disorders.
Miklic
Spela
S
Department of Pharmacology and Experimental Toxicology, Faculty of Medicine, University of Ljubljana, Korytkova 2, SI-1000 Ljubljana, Slovenia.
Juric
Damijana Mojca
DM
Carman-Krzan
Marija
M
Caman-Krzan
Marija
M
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism*; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects / Q000378:metabolism*; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D004298:Dopamine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006706:Homeostasis / Q000187:drug effects / Q000502:physiology; D007608:Kainic Acid / Q000494:pharmacology; D020932:Nerve Growth Factor / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014018:Tissue Distribution; D010959:Tissue Plasminogen Activator / Q000494:pharmacology
D019208:Brain-Derived Neurotrophic Factor; D016207:Cytokines; D005576:Colforsin; D020932:Nerve Growth Factor; D010959:Tissue Plasminogen Activator; D007608:Kainic Acid; D004298:Dopamine