Progress Report U01 014829 Aims 1 & 2: To test L-NAP and other compounds for therapeutic potential in preventing alcohol-induced damage to the cerebellum in a neonatal rat model of 3rd-trimester alcohol exposure during binge drinking Accomplishments: Vitamin E fails to protect against neontal alcohol-induced cerebellar cell death or functional deficits, assessed with caspase-3 activation, cell counts and eyeblink conditioning {published in ACER} L-NAP fails to protect against activation of caspase-3 or loss of cerebellar Purkinje cells Binge Alcohol on PD 4-9 induces dose-dependent deficits on trace eyeblink conditioning

Activation of Caspase-3 in Cerebellar Purkinje Cells (P4)

Progress Report: Binge Neonatal Alcohol Exposure Impairs Trace Eyeblink Conditioning

Progress Report Aim 3: To develop a neonatal mouse model of 3rd-trimester binge exposure and evaluate behavioral correlates of alcohol-induced damage to hippocampal formation and limbic cortex Accomplishments: C57BL/6 mice given heavy binge exposure to alcohol on P7 (2.5 g/kg in each of two injections, 2 hours apart) Activation of caspase-3 in caudate, cingulate cortex, retrosplenial cortex, and hippocampal formation (Zhou) P7 binge exposure did not produce effects opposite that predicted for conditional discrimination reversal task (eyeblink conditioning enhanced responding during discrimination reversal: impaired inhibitory learning?

Ongoing Study and Renewal Trajectory Assess 3-day (P7-P9) binge exposure in B6 mice for enduring consequences of forebrain damage Is there hippocampal – frontal cortical targeting? Is there age-dependent expression? [compare with P7] (as seen by Wozniak et al, 2004 with P7 exposure) Behavioral Studies (tested at P30 and P70) water maze spatial learning Trace Eyeblink Conditioning Anatomical Studies Volumetric and cell count studies of hippocampal formation (adults tested above) Screening for Neuroprotection (with Zhou) caspase-3 activation (forebrain) Behavior Cell counts

What Should the Consortium Basic Science [Animal Models] Do? Use two different C57BL/6 mouse models of alcohol exposure to screen therapeutic agents Implement animal model behavioral tasks that are directly applicable to human population Identify effects in B6 mice; confirm in rat model when needed Prenatal Exposure (liquid diet starting on E7) L-NAP protection works for embryonic brain! Extend over development; identify mechanism(s) [Miller] Need adult testing to screen for long-term protection Binge Neonatal Exposure (injection on P7-9) Currently comparing effects of P7 vs. P7-9 on “hippocampal-dependent” tasks Screen potential agents: Vitamin B3 (nicotinamide) may be a good candidate protective agent (Ierec & Herrera, SfN, 2005)

What Could the Consortium Basic Science [Animal Models] Do? Match assessment tasks in rodents to ones that have parallels in humans (and can be or are used in the consortium) Eyeblink (trace vs. delay; conditioned inhibition) Spatial navigation / spatial memory Fear / avoidance conditioning (behavioral & autonomic) Response to novelty Impulsivity? delay discounting task (insensitivity to future or unpredictable consequences) Circadian regulation? HPA regulation / stress reactivity (CORT)

What Could the Consortium Basic Science [Animal Models] Do? L-NAP Protection against prenatal exposure deficits in B6 Mouse Model (liquid diet beginning on E7) Behavioral Studies (testing L-NAP protection) Develop Behavioral Screening Battery for mice (in collaboration with Stanton/JHU Center for Neurogenetics) Spatial navigation & spatial working memory Eyeblink Conditioning: Trace vs. Delay Eyeblink Condiitioning: Conditioned Inhibition Fear conditioning (contextual vs. signaled) Fear conditioning (trace vs. delay) Activity and response to novelty Prepulse inhibition of acoustic startle Tests of impulsivity (delay discounting task)? Anatomical Studies (adults tested above) with Zhou Find mechanisms of protection

What Could the Consortium Basic Science [Animal Models] Do? Pursue agents that may prevent neonatal damage Screening approach : outcome measures matched to consortium Candidate therapeutics in depth (e.g., vitamin B3) Pursue post-treatment interventions that may help ameliorate the neonatal alcohol-induced damage (esp. targeting cerebellar, hippocampal, or frontal cortical function) Potential interventions Exercise, targeted training, other “enrichment” (currently outside CIFASD) Choline Supplementation (currently outside CIFASD) Pharmacological treatment (diffusely done across the field) Biomarkers!