title: Cyclic adenosine monophosphate and brain-derived neurotrophic factor decreased oxidative stress and apoptosis in developing hypothalamic neuronal cells: role of microglia.
abstract: BACKGROUND
  We have previously shown that ethanol (EtOH) increases cellular apoptosis to developing neurons via the effects on oxidative stress of neurons directly and via increasing production of microglia-derived factors. To study further the mechanism of EtOH action on neuronal apoptosis, we determined the effects of 2 well-known PKA activators, dibutyryl cAMP (dbcAMP) and brain-derived neurotrophic factor (BDNF), on EtOH-activated oxidative stress and apoptotic processes in the hypothalamic neurons in the presence and absence of microglial cells' influence.
  
  
  METHODS
  In enriched neuronal cells from fetal rat hypothalami treated with EtOH or with conditioned medium from EtOH-treated microglia, we measured cellular apoptosis by the free nucleosome assay and the levels of cAMP, BDNF, O²⁻, reactive oxygen species (ROS), nitrite, glutathione (GSH), and catalase following treatment with EtOH or EtOH-treated microglial culture conditioned medium. Additionally, we tested the effectiveness of dbcAMP and BDNF in preventing EtOH or EtOH-treated microglial conditioned medium on cellular apoptosis and oxidative stress in enriched hypothalamic neuronal cell in primary cultures.
  
  
  RESULTS
  Neuronal cell cultures following treatment with EtOH or EtOH-activated microglial conditioned medium showed decreased production levels of cAMP and BDNF. EtOH also increased apoptotic death as well as oxidative status, as demonstrated by higher cellular levels of oxidants but lower levels of antioxidants, in neuronal cells. These effects of EtOH on oxidative stress and cell death were enhanced by the presence of microglia. Treatment with BDNF or dbcAMP decreased EtOH or EtOH-activated microglial conditioned medium-induced changes in the levels of intracellular free radicals, ROS and O²⁻, nitrite, GSH, and catalase.
  
  
  CONCLUSIONS
  These data support the possibility that EtOH by acting directly and via increasing the production of microglial-derived factors reduces cellular levels of cAMP and BDNF to increase cellular oxidative status and apoptosis in hypothalamic neuronal cells in primary cultures.
authors.affiliation: Endocrine Program, Department of Animal Sciences, Graduate Program of Neuroscience, Rutgers, The State University of New Jersey, New Brunswick, NJ, USA.
authors.forename: Nadka I
authors.initials: NI
authors.lastname: Boyadjieva
authors.forename: Dipak K
authors.initials: DK
authors.lastname: Sarkar
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis / Q000276:immunology; D019208:Brain-Derived Neurotrophic Factor / Q000502:physiology*; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000009:adverse effects; D000242:Cyclic AMP / Q000502:physiology*; D000431:Ethanol / Q000009:adverse effects / Q000378:metabolism*; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders / Q000209:etiology; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism*; D017628:Microglia / Q000187:drug effects / Q000502:physiology*; D018384:Oxidative Stress / Q000276:immunology; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation / Q000502:physiology
chemical_list: D000975:Antioxidants; D019208:Brain-Derived Neurotrophic Factor; D002492:Central Nervous System Depressants; D000431:Ethanol; D000242:Cyclic AMP
keywords: Apoptosis; BDNF; Microglia; Neurons; Oxidative Stress; cAMP
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