Charles R.Goodlett, Principal Investigator/Program Director
Testing FASD Therapeutic Agents: Neonatal Rodents Models
Feng C. Zhou, Investigator
Testing FASD Therapeutic Agents in C57/BL Mouse Model
Testing FASD Therapeutic Agents: Neonatal Rodents Models
The long-term goals of this component are to use rodent models of binge alcohol exposure during the 3rd trimester equivalent to screen and identify molecular agents that may be effective in preventing prenatal alcohol-induced brain damage and neurodevelopmental disorders. These studies will use two neonatal rodent models. The first is the well-studied model involving binge alcohol exposure on postnatal days (PD) 4-9 in outbred rats, which is known to produce structural and functional damage to cerebellar systems. The second is a model involving a single binge alcohol exposure on PD7 in the C57BL/6 inbred mouse strain (the focus of Zhou's mouse component), known to induce extensive forebrain neuronal cell death. These studies will assess the effectiveness of candidate therapeutic agents by evaluating alcohol-induced damage to cerebellar (rat) and hippocampal (mouse) circuits that are known to mediate specific variants of Pavlovian eyeblink classical conditioning (ECC). In keeping with the emphasis of the Core, L-NAP, a 9-amino acid derivative of activity-dependent neuroprotective protein, will be the first candidate agent tested, because of its demonstrated effectiveness in several models of cell death and in a mouse model alcohol induced teratogenesis. If L-NAP is found to be effective, additional studies of two structural derivatives of L-NAP for which the capacity to protect against neuronal death vs. ethanol teratogenesis has been dissociated, will also be evaluated. Specific Aim 1 tests the hypothesis that the candidate agent will protect against alcohol-induced caspase-3 activation on PD 4 in Purkinje cells, an indicator of acute Purkinje cell death. Specific Aim 2 tests the hypothesis that the candidate agent will protect against functional and structural damage to cerebellar systems mediating eyeblink conditioning induced by binge alcohol exposure on PD 4-9. Specific Aim 3 tests the hypothesis that binge alcohol exposure on PD 7 in C57BL/6 mice will damage hippocampal circuits necessary for eyeblink discrimination reversal learning, and that the candidate agent will protect against this hippocampal damage. Because the neural substrates and procedures for ECC are similar across species, the outcomes from these animal models including those of the sheep model component of this Consortium can be translated directly to humans to guide and inform future studies of therapeutic prevention of FASD. A key advantage of integrated approaches across the Consortium is that as promising candidate molecular agents emerge, these animal models can provide in vivo tests of their therapeutic effectiveness.
Testing FASD Therapeutic Agents in C57/BL Mouse Model
Maternal alcohol consumption is the leading known cause of mental retardation in the Western world. The fetal alcohol spectrum disorder (FASD) including fetal alcohol syndrome and alcohol-related neurodevelopmental disorder causes a life-time disability to the victim. The goal of this project is to test therapeutic agents that can protect against FASD, particularly the central nervous system. The activity dependent trophic factor and their associated peptides has been known to be extremely potent as protectant against oxidative stress and alcohol induced fetal demise.
The goals of this project will be to (a) test the protective/neurotrophic effects of ADNF-like peptides against alcohol compromised neurons, particularly the serotonin, and excitatory neurons in the limbic system. Due to extremely dynamic change in neural development, the effect of ADNF-like peptides will be tested against alcohol exposure at pre- and neo-natal stages of mice equivalent to the human lst/2nd and 3rd trimesters respectively. (b) Second, we will study the mechanism of ADNF-like peptide protection by analyzing its gene expression in a selective paradigm.
The Aim 1 will test the neuroprotective and/or neurotrophic effect of ADNF-like peptides against alcohol neurotoxicity to developing serotonin (5-HT) and hippocampal neurons by study their apoptosis, number of matured neurons, and degree of 5-HT and glutamate innervation in the hippocampus with an established alcohol liquid diet model at E7- E18, equivalent to 1st and 2nd trimesters of human gestation. The Aim 2 will determine the neuroprotective and neurotrophic effect of ADN F-like peptide on an alcohol induced apoptosis, neurogenesis, synaptogenesis of the limbic system with binge ethanol exposure at a neonatal stage equivalent to 3rd trimester of human gestation. The Aim 3 will analyze the mechanism of ethanol neurotoxicity in the neonatal binge alcohol exposure model, and the protective effects of ADNF-like peptide upon gene expression, for understanding toxicity and protection, using microarray analysis, real-time RT-PCR and in situ hybridization.