title: Dysregulation of TrkB phosphorylation and proBDNF protein in adenylyl cyclase 1 and 8 knockout mice in a model of fetal alcohol spectrum disorder.
abstract: Brain-derived neurotrophic factor (BDNF) mediates neuron growth and is regulated by adenylyl cyclases (ACs). Mice lacking AC1/8 (DKO) have a basal reduction in the dendritic complexity of medium spiny neurons in the caudate putamen and demonstrate increased neurotoxicity in the striatum following acute neonatal ethanol exposure compared to wild type (WT) controls, suggesting a compromise in BDNF regulation under varying conditions. Although neonatal ethanol exposure can negatively impact BDNF expression, little is known about the effect on BDNF receptor activation and its downstream signaling, including Akt activation, an established neuroprotective pathway. Therefore, here we determined the effects of AC1/8 deletion and neonatal ethanol administration on BDNF and proBDNF protein expression, and activation of tropomyosin-related kinase B (TrkB), Akt, ERK1/2, and PLCγ. WT and DKO mice were treated with a single dose of 2.5 g/kg ethanol or saline at postnatal days 5-7 to model late-gestational alcohol exposure. Striatal and cortical tissues were analyzed using a BDNF enzyme-linked immunosorbent assay or immunoblotting for proBDNF, phosphorylated and total TrkB, Akt, ERK1/2, and PLCɣ1. Neither postnatal ethanol exposure nor AC1/8 deletion affected total BDNF protein expression at any time point in either region examined. Neonatal ethanol increased the expression of proBDNF protein in the striatum of WT mice 6, 24, and 48 h after exposure, with DKO mice demonstrating a reduction in proBDNF expression 6 h after exposure. Six and 24 h after ethanol administration, phosphorylation of full-length TrkB in the striatum was significantly reduced in WT mice, but was significantly increased in DKO mice only at 24 h. Interestingly, 48 h after ethanol, both WT and DKO mice demonstrated a reduction in phosphorylated full-length TrkB. In addition, Akt and PLCɣ1 phosphorylation was also decreased in ethanol-treated DKO mice 48 h after injection. These data demonstrate dysregulation of a potential survival pathway in the AC1/8 knockout mice following early-life ethanol exposure.
authors.affiliation: John D. Dingell VA Medical Center, Wayne State University School of Medicine, Detroit, MI 48201, USA; Department of Neurosurgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Laura L
authors.initials: LL
authors.lastname: Susick
authors.affiliation: John D. Dingell VA Medical Center, Wayne State University School of Medicine, Detroit, MI 48201, USA; Department of Neurosurgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Alexandria C
authors.initials: AC
authors.lastname: Chrumka
authors.affiliation: John D. Dingell VA Medical Center, Wayne State University School of Medicine, Detroit, MI 48201, USA; Department of Neurosurgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Hool
authors.affiliation: John D. Dingell VA Medical Center, Wayne State University School of Medicine, Detroit, MI 48201, USA; Department of Neurosurgery, Wayne State University School of Medicine, Detroit, MI 48201, USA. Electronic address: aconti@med.wayne.edu.
authors.forename: Alana C
authors.initials: AC
authors.lastname: Conti
mesh_terms: D000262:Adenylyl Cyclases / Q000172:deficiency / Q000378:metabolism*; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism*; D000431:Ethanol / Q000633:toxicity; D063647:Fetal Alcohol Spectrum Disorders / Q000209:etiology / Q000235:genetics / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010766:Phosphorylation / Q000187:drug effects / Q000502:physiology; D011498:Protein Precursors / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D020813:Receptor, trkB; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D008562:Membrane Glycoproteins; D011498:Protein Precursors; C426674:brain-derived neurotrophic factor precursor; D000431:Ethanol; D011505:Protein-Tyrosine Kinases; D020813:Receptor, trkB; C000591259:tropomyosin-related kinase-B, human; D000262:Adenylyl Cyclases; C106670:adenylyl cyclase 1; C118292:adenylyl cyclase 8
keywords: Akt; Brain-derived neurotrophic factor; Fetal alcohol spectrum disorder; Striatum; Tropomyosin-related kinase B