Prenatal Alcohol Exposure: Neuropathology and cognition Integrating Behavioral & Neural

1. Prenatal Alcohol Exposure: Neuropathology and cognition Integrating Behavioral & Neural Approaches in Rats and Humans Rob Sutherland Canadian Centre for Behavioural Neuroscience The University of Lethbridge

2. Project Directors Robert Sutherland Dan Savage Derek Hamilton MEG Claudia Tesche Kim Paulson Sandra Moses Rat Model Rob McDonald Linda Beyer-Smith Rafael Galindo Janice Hoesing Michael Thomas Ying Wu Patti Sorensen Subject Recruitment Carol Clericuzio Luther Robinson Julia Powers Jorge Ganem Hannah Olivet sMRI, MRS & fMRI Roland Lee Allison Lindsay William Brooks Helen Petropoulos Neuropsychological Assessment P.W. Kodituwakku Barbara Wilson Paula Wilbourne

3. Moderate Ethanol Exposure Paradigm • 5% EtOH diet throughout gestation. • Pair-Fed 0% EtOH and Ad Lib controls • 16 Hour feeding / 8 hour water schedule. • No significant effects of the ethanol diet on:- Litter size or Neonatal mortality. - Offspring birthweight or growth curves. - Brain weight or gross anatomy after sacrifice.

4. Morris water task

5. Simple, constant learning tasks are unaffected

6. 20 Trial 1 Trial 2 Latency to find platform (s) 10 0 Ad lib Pair-fed 5% alcohol Group One-trial learning tasks that tax hippocampus are affected

7. Synaptic plasticity in hippocampus

8. Ad Lib 0.8 0.8 Pair-Fed 0.7 0.7 5% EtOH 0.6 0.6 0.5 0.5 fEPSP ( Fractional Change ) 0.4 0.4 0.3 0.3 0.2 0.2 0.1 0.1 0.0 0.0 0 20 40 60 80 100 Time ( minutes ) Synaptic plasticity is impaired

9. A. Pair-fed to 3% EtOH 2.5 3% EtOH Diet of D-ASP (%) 100 2.0 Fractional Release 80 60 1.5 Concentration (mg/dL) Maternal Blood Ethanol S1 TET S2 40 0 6 12 18 24 84 90 96 102 108 Fraction Intervals (minutes) 20 B. 0 2% 3% 5% 1.4 Liquid Diet Ethanol Concentration * Release (S2 / S1) * * 1.2 Potentiation of D-ASP 1.0 Ad PF-2 2% PF-3 3% PF-5 5% Lib EtOH EtOH EtOH Threshold for adult synaptic effect is .03 BAL

10. B. 14 * 12 10 8 Improvement in * Escape Latency (Sec) 6 ** 4 * 2 * 0 Ad PF-2 2% PF-3 3% PF-5 5% Lib EtOH EtOH EtOH 3% maternal alcohol affects adult learning

11. Green = neuron Red = new cell Gold = new neuron Adult hippocampus has thousands of new cells each day. Alcohol exposed rats have many fewer new neurons in adult hippocampus

12. Visual Water Task Correct Response Incorrect Response Monitors Platform Divider Pool Choice Plane

13. Visual acuity 1 0.9 0.8 0.7 0.6 Correct 0.5 0.4 Ad lib 0.3 Pair-fed 5%EtOH 0.2 0.1 0 0.564 0.1187 0.1484 0.1781 0.2078 0.2375 0.2671 0.2968 0.3562 0.3859 0.4156 0.4452 0.4749 0.5343 0.3265 0.5046 0.6233 0.5937 Spatial frequency Visual accuity not affected.

14. Fetal Alcohol - Induced Deficits • Hippocampus-sensitive memory tasks • Hippocampal synaptic plasticity • Neurochemical mechanisms associated with synaptic plasticity • Threshold is around .03 BAL for adult synaptic and learning deficits • Adult neurogenesis is decreased

15. What about people? Neuroimaging in FAS

16. A virtual Morris water task

17. A virtual Morris water task Phase I: Place learning (Twenty trials) Phase II: Probe trial Phase III: Cued-navigation (Eight trials)

18. VMWT Results Virtual navigation is impaired. Examples of swim paths to hidden platform

19. VMWT Results Searching in correct region on probe is impaired. Dwell time in regions of the pool

20. VMWT Results Virtual navigation to a cue is unaffected. Examples of swim paths to visible platform

21. VMWT and Neuroimaging Day 1 Neuropsychological test battery Train subjects on the VMWT MEG/EEG recording while performing VMWT Day 2 sMRI Proton MRS (right HPC & right DLPFC) fMRI recording while performing VMWT.

22. Virtual navigation with beacons. On different trial blocks participants must learn which cue is correct or which location is correct

23. Magnetoencephalography

24. Front Magnetoencephalography

25. Magnetoencephalography

26. Magnetoencephalography

27. MEG Results Right hippocampal activation during place learning was observed in 2/7 children with FAS Right hippocampal activation during place learning was observed in 6/7 controls

28. MEG Results and Behavior 80 70 60 50 Initial Heading Error (deg) 40 30 20 10 0 + - Right Hippocampal ECD

29. Structural MRI An automated procedure separates white and grey matter from sMRIs and calculates volume of hippocampal grey matter.

30. Structural MRI Total hippocampal volume is unaffected.

31. Magnetic Resonance Spectroscopy MRS permits quantification of several neurochemicals from regions of prefrontal cortex and hippocampus.

32. Magnetic Resonance Spectroscopy 1 H-Magnetic Resonance Spectroscopy 3.0 CONTROL 2.5 FAS 2.0 NAA / Creatine 1.5 1.0 0.5 0.0 Hippocampal DL Prefrontal Formation Cortex No significant differences in neurochemistry**

33. Human Summary Place learning in the VMWT Participants with FAS were impaired at hippocampal- dependent place learning in the VMWT Morphometrics/Volumetrics (sMRI) No evidence for alcohol-related differences in hippocampal volume/structure Neurometabolites (1H-MRS) No evidence for alcohol-related differences in hippocampal neurometabolite profiles Functional Neuroimaging (MEG) Participants with FAS were less-likely to display task-related activity in right hippocampus.Task-related activity in HPC was highly correlated with behavior in the VMWT

34. Conclusions • Alcohol-related differences in brain function can be detected using technology that is currently available • Alcohol-related differences in brain function may not correspond with neuroanatomical differences detected using structural MRI or MRS • Functional techniques may be particularly useful and sensitive for identifying the neural bases of cognitive dysfunction in FASD • Functional neuroimaging may help provide unique diagnostic information and may be helpful in assessing treatment • It will be important to relate functional brain activation with behaviorally-relevant paradigms – Linking basic human and animal research on FASD with clinical research