Cellular and Genetic Factors in Traumatic Brain Injury

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Acknowledgments

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Table of Contents 1. Title i 2. Acknowledgements ...ii 3. Table of Contents.iii 4. List of Figures and Graphs ............................v 5. Abstract vii 6. Introduction .1 6.1 Introduction to Traumatic Brain Injury..1 6.2 Microglias Function and role in TBI .... 6.3 Astrocytes Function and role in TBI.....7 6.4 ApoE and its relation to TBI.8 7. Methods and Procedures.38 7.1 Introduction .38 7.2 Experiments 7.2a Apo e 133 149 .38 7.2b APP vs. WT 39 7.2c 2 vs. 4 .. 39 7.2d TBI Severity Study.......40 7.3 General Overview .40

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7.3a TBI infliction.. 40 7.3b Fixation, Embedding and Sectioning 7.3c Staining Techniques .41 7.4d Quantification and Statistical Analysis ..43 8. Results...44 8.1 ApoE 133 149 44 8.1a Microglia Analysis ...44 8.1b Astrocyte Analysis ..45 8.2 APP vs. WT 8.2a 4G8 Analysis 8.2b Astrocyte Analysis .....47 8.3 2 vs. 4 ..47 8.3a Microglia Analysis ...48 8.3b Astro cyte Analysis ..49 8.4 TBI Severity Study ..49 8.4a Astrocyte Analysis ..49 9. Conclusion..52 10. Bibliography.. 54

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List of Figures and Graphs Figures Figure 1 .1 Figure 2 .6 Figure 3 ..10 Figure 4 ..12 Figure 5 ..13 Figure 6 ..15 Figure 7 ..16 Figure 8 ..18 Figure 9 ..19 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Fig ure 17

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Figure 18 Figure 19 Figure 20 Figure 21 Graphs Graph 1 ...44 Graph 2 ...45 Graph 3 ...46 Graph 4 ...47 Graph 5 ...48 Graph 6 ...49 Graph 7 ...50

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1. Introduction 1.1 Introduction to Traumatic Brain Injury Figure 1 Figure 1.

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Figure 2.

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Figure 2. a c d e d e f g f g 1.2 Microglias functions and role in traumatic brain injury

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1.3 Astrocytes function and role in traumatic brain injury

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1.4 ApoE and its relation to TBI

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Figure 3 Figure 3. a) b) c) d) Figure 3d

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p p p Figure 4

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Figure 4. Figure 5

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Figure 5. A) B) C) D)

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p p

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Fig. 6 Figure 7. Figure 6.

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Figure 7. A) B) C) p

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Figure 8 p < p

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Figure 8 p t in vivo Figure 9

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in vitro Figure 9. in vivo Figure 10 in vivo in vitro

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F igure 10. In vivo

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p Figure 11a Figure 11b

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Figure 11. a) b) p<

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Figure 12 Figure 12. p

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Figure 13a Figure 13b Figure 14

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Figure 13. a) p b) Figure 14

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Figure 15. Figure 15. A) B) p

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p p Figure 16

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Figure 16.

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Figure 17 Figure 18 NH CH3CH2O-O OH O Figure 17. Figure 18.

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p p<. Figure 19

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Figure 19. p Figure 20

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a) b) Figure 20. a) b)

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dementia pugilistica Figure 21

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Figure 21.

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2. Methods and Procedures2.1 Introduction 2.2 xperiments E2.2a ApoE 133 149

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2.2b APPvs. WT 2.2c. 2 vs. 4

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2.2d Traumatic Brain Injury Severity Study 2.3 General Overview 2.3.a TBIInfliction 2.3b Fixation, Embedding and Sectioning

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2.3c Staining Techniques

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2.4d Quantification and Statistical Analysis P

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3. Results 3.1 ApoE 133 149 3.1a Microglial Analysis Graph 1. Graph1. P p p

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3.1b Astrocyteanalysis Graph 2. Graph 2. P ShamVehicle Cortex vs TBI Vehicle Cortex p <0.01 ShamVehicle Cortex vs TBI Vehicle Hippocampus p <0.001 ShamVehicle Thalamus vs TBI Vehicle Cortex p <0.01 ShamVehicle Thalamus vs TBI Vehicle Hippocampus p <0.001 ShamVehicle Hippocampus vs TBI Vehicle Cortex p <0.01

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ShamVehicle Hippocampus vs TBI Vehicle Hip p <0. 001 ShamApoE Cortex vs TBI Vehicle Cortex p <0.01 ShamApoE Cortex vs TBI Vehicle Hippocampus p <0.001 ShamApoE Thalamus vs TBI Vehicle Cortex p <0.01 ShamApoE Thalamus vs TBI Vehicle Hippocampus p <0.001 ShamApoE Hippocampus vs TBI Vehicle Cortex p <0.05 SgamApoE Hippocampus vs TBI Vehicle Hippocampus p <0.001 TBI Vehicle Hippocampus vs TBI ApoE Cortex p <0.05 TBI Vehicle Hippocampus vs TBI ApoE Thalamus p <0.01 TBI Vehicle Hippocampus vs TBI ApoE Hippocampus p <0.01 3.2APP vs. WT 3.2a 4G8 positive plaquecount Graph 3. Graph 3 P

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3 .2b Astroc ytelevels b) c) Graph 4. a) b) c) P p 3 3 .3 2 vs. 4.3a Micro glial analysis

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a) b)Graph 5. a) b) Graphs 5a 5b P p

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3 .3b GFAP anal ysis Graph 6. Graph 6 P p 3.4Traumatic Br ain Injury Severity Study 3.4a Astrocyte 2 weekand 4 weekanalysis

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a) b) Graph 7. a) b) Graphs 7 P

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2week Sham Cortex vs 4week TBI 1.3mm Cortex P<0.01 2week Sham Cortex vs 4week TBI 2.0mm Cortex P<0.05 2week Sham Hippocampus vs 4week TBI 1.3mm Cortex P<0.05 2week Sham Hippocampus vs 4week TBI 2.0mm Cortex P<0.05 2week TBI 1.3mm Hippocampus vs 4week TBI 1.3mm Cortex P<0.05 4week TBI 1.3mm Cortex vs 4week Sham Hippocampus P<0.01 4week TBI 1.3mm Cortex vs 4week TBI 1.3mm Hippocampus P<0.01 4week TBI 1.3mm Cortex vs 4week TBI 2.0mm Hippocampus P<0.05 4week TBI 2.0 mm Cortex vs 4week Sham Hippocampus P<0.05 4week TBI 2.0mm Cortex vs 4week TBI 1.3mm Hippocampus P<0.05 4week TBI 2.0mm Cortex vs 4week TBI 2.0 mm Hippocampus P<0.05

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4. Conclusion

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5. Bibliography

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