title: Chronic ethanol treatment and withdrawal alter ACPD-evoked calcium signals in developing Purkinje neurons.
abstract: BACKGROUND
  Alcohol exposure during human fetal development can result in fetal alcohol syndrome, a condition characterized by central nervous system dysfunction. Detailed studies in animal models of fetal alcohol syndrome show that the cerebellar region is particularly sensitive to alcohol exposure during early development; however, the cellular mechanisms underlying the alcohol sensitivity of the immature cerebellum are poorly understood.
  
  
  METHODS
  Primary neuronal cultures of cerebellar cells were prepared from embryonic day 20 rat pups. Cultures were exposed to ethanol (33 mM; 150 mg/100 ml) during the main period of morphological development of the Purkinje neurons, from 6 to 17 days in vitro. After the ethanol treatment, the response of Purkinje neurons to the selective metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD; 300 microM) was examined in parallel fura-2 Ca2+ imaging and current-clamp experiments. In an additional set of experiments, ethanol-treated cultures were allowed to withdraw from ethanol overnight before recordings were performed.
  
  
  RESULTS
  In Ca2+ imaging studies, the mean peak amplitude of ACPD-evoked Ca2+ signals was depressed in the dendritic region of chronic ethanol-treated Purkinje neurons compared with control neurons (p < 0.05, unpaired t test), whereas there was no apparent difference in the somatic region. In contrast, peak ACPD-evoked Ca2+ signals were enhanced in both the somatic and dendritic regions of withdrawn Purkinje neurons compared with control neurons. Parallel current-clamp studies showed no consistent effect of chronic ethanol treatment or ethanol withdrawal on the membrane response to ACPD.
  
  
  CONCLUSIONS
  These results show that prolonged ethanol exposure and early withdrawal lead to alterations in mGluR-evoked Ca2+ signaling in cerebellar Purkinje neurons. Metabotropic GluRs in the Purkinje neuron play important roles in cerebellar development and function, suggesting that alterations of mGluR signaling pathways by ethanol may play a key role in the actions of ethanol on the developing cerebellum.
authors.affiliation: Department of Neuropharmacology, Alcohol Research Center, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, CA 92037, USA.
authors.forename: Jeffrey G
authors.initials: JG
authors.lastname: Netzeband
authors.forename: Jaimes R
authors.initials: JR
authors.lastname: Schneeloch
authors.forename: Carol
authors.initials: C
authors.lastname: Trotter
authors.forename: Jodilyn N
authors.initials: JN
authors.lastname: Caguioa-Aquino
authors.forename: Donna L
authors.initials: DL
authors.lastname: Gruol
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects* / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000008:administration & dosage / Q000009:adverse effects*; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000196:embryology; D003515:Cycloleucine / Q000031:analogs & derivatives / Q000494:pharmacology*; D004622:Embryo, Mammalian; D000431:Ethanol / Q000008:administration & dosage / Q000009:adverse effects*; D011689:Purkinje Cells / Q000166:cytology / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018094:Receptors, Metabotropic Glutamate / Q000819:agonists / Q000502:physiology; D013375:Substance Withdrawal Syndrome / Q000378:metabolism*
chemical_list: D002492:Central Nervous System Depressants; D018094:Receptors, Metabotropic Glutamate; D003515:Cycloleucine; C036361:1-amino-1,3-dicarboxycyclopentane; D000431:Ethanol