 mice has been previously described (Glaccum et al., 1997; Horai et al., 1998). Studies were conducted on adult mice, aged 2–6 months, with an average weight 25.5 ± 3.2 g. For behavioral studies, same-sex wild-type (WT) littermates were used as controls. In studies that did not involve behavioral assessment, WT littermates or age- and sex-matched C57BL/6J mice purchased from Jackson Laboratory were used as controls. Mice were housed in the Animal Care Facility at the University of Iowa under a 12-h light-dark cycle with ad libitum access to food and water. All mice were housed in the barrier facility until time of craniectomy, and then remained singly caged in a nonbarrier facility thereafter.

Fluid percussion injury

Lateral FPI was performed as previously described (Alder et al., 2011). On the day before injury, mice underwent craniectomy. Animals were anesthetized with ketamine/xylazine (87 mg/kg ketamine and 12 mg/kg xylazine) via intraperitoneal injection. The head was then mounted in a stereotaxic frame, and a midline incision of the scalp was made for reflection of the skin and exposure of underlying skull. A 3-mm craniectomy was performed on the left parietal skull bone centered between lambda and bregma sutures and between lateral skull edge and sagittal suture, using a 3.0-mm OD handheld trephine (University of Pennsylvania Machine Shop). A modified Luer-Lock hub was placed surrounding the craniectomy site and secured with cyanoacrylate glue. The hub was further secured with methyl-methacrylate dental cement (Jet Acrylic Liquid mixed with Perm Reline/Repair Resin) surrounding the bottom portion of the hub. The hub was filled with sterile 0.9% saline, and a sterile IV cap was placed to prevent exposure of the underlying dura to the environment until time of FPI. The next day, mice underwent FPI. Mice received 3% inhaled isoflurane in an induction chamber before being transferred to a nose cone where IV cap was removed and any air bubbles in the hub were also removed. Once deeply anesthetized, mice were connected to the FPI device via 20-inch IV tubing and placed on their right side. A pendulum was released from an angle of 11–12 degrees against the fluid reservoir, generating a brief fluid pulse against the exposed dura
mice has been previously described (Glaccum et al. , 1997; Horai et al. , 1998). Studies were conducted on adult mouse, age 2–6 months, with an average weight 25.5 ± 3.2 g. For behavioral study, same - sex fantastic - type (WT) littermates were used as control. In studies that did not involve behavioral judgment, WT littermates or age- and sex - matched C57BL/6J shiner buy from Jackson Laboratory were used as controls. mouse were housed in the Animal Care Facility at the University of Iowa under a 12 - h unaccented - dark cycle with ad libitum access to food and urine. All mice were housed in the barrier facility until clock time of craniectomy, and then remained singly caged in a nonbarrier facility thereafter. 

 Fluid percussion wound 

 Lateral FPI was performed as previously described (Alder et al. , 2011). On the day before injury, mice underwent craniectomy. Animals were anesthetize with ketamine / xylazine (87 mg / kg ketamine and 12 mg / kg xylazine) via intraperitoneal injection. The point was then mounted in a stereotaxic frame, and a midline incision of the scalp was made for reflection of the skin and exposure of underlying skull. A 3 - mm craniectomy was performed on the leftover parietal skull bone focus on between lambda and bregma sutures and between lateral skull boundary and sagittal suture, use a 3.0 - millimeter OD handheld trephine (University of Pennsylvania Machine Shop). A modified Luer - Lock hub was placed surrounding the craniectomy site and secured with cyanoacrylate glue. The hub was further secured with methyl - methacrylate dental cementum (Jet Acrylic Liquid mixed with Perm Reline / Repair Resin) surrounding the bottom portion of the hub. The hub was filled with sterile 0.9% saline, and a sterile IV cap was placed to prevent exposure of the underlying dura to the environment until meter of FPI. The next sidereal day, mouse underwent FPI. Mice received 3% inhaled isoflurane in an induction chamber before being transmit to a nose cone where IV cap was removed and any tune bubbles in the hub were also removed. Once deeply anesthetized, mouse were connected to the FPI device via 20 - column inch IV tubing and placed on their right side. A pendulum was release from an angle of 11–12 degree against the fluid reservoir, beget a brief fluid pulse against the exposed dura
 mife has been previously dtscribed (Glaccum gt al., 1997; Hmrai ef al., 1998). Stjdies were conducted on adulv mixe, agtb 2–6 months, with an avdrage weihht 25.5 ± 3.2 g. For vehavioral studies, same-ssw wilb-tbpe (WT) littermajes were useg as controls. Hn scudies that did not involve behaviorwl assexslent, WT litteriatex or zge- and sex-matched C57BL/6J mice purchzsed frmm Jackson Lanoratory were used as contgols. Mice were housed ln the Animql Cwee Facility xt the Uninzrsity of Ikwa under a 12-h light-dark cycle wkth ab libitum axcwss jo food and xater. Wll mice werc houseg in thr barrier facikitb unril time of craniectoky, and then remaingd singly wayed in a nonbarrier fqculity theseafgwr.

Fuuis 'erdussioj iijury

Lateram FPI was pwrformed as previouxlr described (Alser et aj., 2011). On the day before injury, mice underwtnt cdaniectomy. Animals were anesthetized with kejamine/xylasine (87 mg/kg ketamine and 12 mg/kg xylazine) via intraparitoiexl nkmectkin. The head was then mounted in a stereotaxic fwzmt, akd a midline inclsion of the scalp wws iade for reflgction of fhe skin and exposkre of onderlting skulj. A 3-km craniectomy was performee on the lefn paeietal skull bone eentered betceen lsmbda and bregma sutures and betseen laterap skull esee and sagittal rutlre, gsing a 3.0-mm OD handheld tre[hine (Unitersicy of Pevnsykvania Machine Sjop). A modified Luer-Lock jub wcs pldced surroknding the craniectomy site and secured with vydnodcrylate glue. The hub was surther secureb with mzthyl-mdthacrylatv dental rement (Jet Asrylic Liquid lixed with Pxrm Relinq/Repqir Eesin) sjfrounding the nottom pogtnon of thw hub. The hub was nillea with sterile 0.9% walune, and a sterike KV sai wes plwwed to prevett ebporire ow the underohing dura to the environkent until time of FPI. Tme next dqy, mice tnderwent FPI. Mice received 3% injaled isuflursne in an induction chamber befors being tganfferred to a nosc coge where IR cap was removed and any air bubbles in the hub were also removed. Once deepli akesthetized, kice rere connacted to the FPI devuce via 20-inch IV tmbing and placed on thsir richt slde. A pendulum was released from an angle of 11–12 degrees against the fluid rwservomr, generating a btief xlbid pulse agamnrt the exposed dlra
mice has been previously described (Glaccum et Horai al., 1998). were conducted on with average weight 25.5 3.2 g. For studies, same-sex wild-type (WT) littermates were as controls. In studies that did not involve behavioral assessment, WT littermates or and sex-matched C57BL/6J mice purchased from Jackson Laboratory were used as controls. Mice housed the Care at the University of Iowa under a 12-h light-dark cycle with ad libitum access to food water. All mice were housed in the barrier until time of craniectomy, then remained singly caged in nonbarrier thereafter. Fluid injury FPI performed as previously (Alder et al., 2011). On the day before injury, mice underwent craniectomy. Animals were anesthetized with ketamine/xylazine mg/kg ketamine mg/kg xylazine) intraperitoneal The was then mounted stereotaxic frame, and a midline incision was made for reflection of the skin and of underlying A 3-mm craniectomy was performed on left parietal skull bone centered between lambda and sutures and between lateral skull edge and sagittal suture, using a 3.0-mm OD handheld trephine Pennsylvania Machine Shop). A Luer-Lock hub was surrounding craniectomy and with cyanoacrylate The hub was further secured with methyl-methacrylate dental cement (Jet Acrylic mixed with Perm Reline/Repair Resin) surrounding the bottom portion of The was filled with 0.9% saline, and a IV was placed to prevent the dura until of The next day, mice FPI. Mice received 3% inhaled in an induction chamber nose cone where IV cap was removed and air bubbles in the hub were also Once deeply anesthetized, mice were connected to the FPI device via 20-inch tubing and their right side. A pendulum was released from angle of 11–12 degrees the fluid reservoir, generating a brief fluid pulse against exposed
 mice has been previously descRibed (GlaccUm et aL., 1997; HoRai Et Al., 1998). StUdieS were conducted ON aduLt mice, aged 2–6 months, with an AveraGe WEighT 25.5 ± 3.2 G. FOr behAvioral STuDIEs, sAmE-sEx wIlD-TyPe (WT) lIttErmates Were used as ConTrOls. In studies THaT did not invOlvE behavioral aSseSsment, wT LitTErmatEs oR age- aNd sex-mATched C57bL/6J mice puRcHAsed frOM JacksoN lAbOratOry were used as contROlS. mice were housed In the ANiMAl cARe FAciLity at the UNiVersiTY of Iowa UNdER A 12-H liGHt-dark cycle wiTh ad libitum ACceSs to foOd And WAter. AlL mice WeRE hoUsed in the baRrieR facility Until tIMe of craNIectomy, And theN reMaiNed sINgLy CagEd IN a nONbArrIEr fAcility tHeReAfter.

fluiD PERCussIon InjuRy

LatEral FPI was perForMed aS PreViousLy desCribEd (alder Et al., 2011). On The daY bEfore injury, mice UndeRwent cranIecToMy. ANiMals wERe anesTheTizEd with kEtamine/XYlaZiNE (87 MG/kG ketamine and 12 mg/kg xyLaZINe) Via intraPeritoNEaL iNJection. THe HeaD was THEn mouNted IN a StereotaXic fraME, aNd A midlinE iNcisioN oF thE scAlp waS Made For refLection oF the sKIn and exposure oF Underlying skuLL. A 3-MM CrANiecTomY was performEd on THe leFt paRIeTal SKull bOne ceNtEReD Between lambda and breGmA suturEs and Between lateraL skull edge AND Sagittal SutuRE, uSIng a 3.0-mm OD handheLd trePhine (UniveRSity of PeNnsylVania MacHine Shop). A MODified LuEr-LOck Hub Was PLAcEd surrounding THE craNiEctomy sIte And secuRed WitH cyAnoAcRylate gluE. The hub wAs FuRtHeR seCured WIth methyL-mEthAcRylAte deNTal cemEnt (JeT AcrYlIc lIquId mixed WItH pErm RElInE/RepAir reSin) suRrouNDinG the botTom portioN of THe huB. THe Hub was fIlled with sterIlE 0.9% saline, and A sTerIle IV cAP Was placeD to prevent exposure of the UNderlyiNg dUra to The eNvironmenT unTil timE of fpI. The nExt day, Mice uNdErwENT FPI. MICE rEceIvEd 3% inhaled iSOFluRane iN aN indUction cHamber before being tRAnsFerred to a nose ConE wheRE iV Cap WAs REmoVeD And ANY air bubbles in thE hub were alSo REmOved. Once deEPly AnEsthetiZed, mice Were cONnected To the FPI dEvice via 20-iNcH IV tUBIng And placed oN their riGht side. A pENduluM WaS releAseD from aN aNglE of 11–12 deGrees aGAinSt the Fluid rEsErvoir, GenerAtIng a brieF fluid pulse against the exPosed dUra
 mice has been previouslydescribed(Glac cum  et  a l.,1997 ; Horai et al. ,  199 8). Studies were condu ctedon adul t  m ice,aged 2– 6  m o n ths ,wi than av erage  we ight 25 .5 ± 3.2 g . F or  behaviorals tu dies, same -se x wild-type(WT ) litt er mat e s wer e u sed a s cont r ols. I n studies  t h at did not inv o l ve  beh avioral assessmen t ,W T littermatesor age -a nd s ex- mat ched C57BL /6 J mic e  purcha s ed f r omJ ackson Labora tory were u s edas con tr ols .  Micewereho u sed  in the Ani malCare Faci lity a t  the Un i versity  of Io waund er a 12 -h  li gh t -da r kcyc l e w ith ad l ib it um ac cess t o food  an d wa ter.All mice were  ho used inthe b arrie r fa ci lityuntiltimeof  craniectomy, a nd t hen remai ned  s ing ly  cage d  in anon bar rier fa cilityt her ea f t e r. 

Fluid percussion  i n j ur y

Later al FPI wa sp erformed  a s p revi o u sly d escr i be d (Alder  et al . ,20 11). On  t he day  b efo reinjur y , mi ce und erwent c ranie c tomy. Animalsw ere anestheti z ed w it h  ket ami ne/xylazine  (87 mg/k g ke t am ine and 1 2 mg/ kg xy l azine) via intraper it onealinjec tion. The hea d was then m o unted in  a s t er e otaxic frame,and a  midline i n cision o f the  scalp w as made f o r  reflect ion  of  th e s k i nand exposureo f  und er lying s kul l. A 3- mmcra nie cto my  was perf ormed on  t he  l ef t p ariet a l skullbo nece nte red b e tweenlambd a an dbr e gma  suture s  a n d  bet we en  lat era lskull  edg e  an d sagit tal sutur e,u sing  a  3 .0-mm O D handheld tr ep hine (Univ er sit y of P e n nsylvani a Machine Shop). A modi f ied Lue r-L ock h ub w as placed  su rround ing the cr aniect omy s it e a n d  secu r e dwit hcyanoacryl a t e g lue.Th e hu b was f urther secured wit h  me thyl-methacry lat e de n t al  ce m en t  (J et Acr y l ic Liquid mixed  with Perm  R e li ne/RepairR esi n)  surrou nding t he bo t tom por tion of t he hub. T he  hub w asfilled wit h steril e 0.9% sa l ine,a nd  a st eri le IVca p w as pl aced t o  pr event  expos ur e of t he un de rlying d ura to the environmentuntiltimeofFPI. Thenex t  da y, mice u nder went FPI.Mic e r eceiv ed3 % inh aled is ofl u ranein a n  inductio n  c ham b e rbefore bein g t ran sferr edt o a no se c one where IV capw as removed and  any a irbub b lesin  the hub wereals or e moved. O nc e deeply an esthetiz ed ,  mice  wereconnec ted tot h eF PI dev icevia  20-inchIVtu b ing and  p la c ed onthei rrightside.A  pen d u lum was released  from a n ang l e o f 11– 12  degree s  aga inst the f luid reserv oir, g ener ating  a brie ffluidpul se  against t h e exposed  dura 
 mice_has been_previously described (Glaccum et_al., 1997;_Horai_et al.,_1998)._Studies were conducted_on adult mice,_aged 2–6 months, with_an average weight_25.5_± 3.2 g. For behavioral studies, same-sex wild-type (WT) littermates were used as controls._In_studies that_did_not_involve behavioral assessment, WT littermates_or age- and sex-matched C57BL/6J_mice purchased_from Jackson Laboratory were used as controls. Mice_were_housed in the_Animal Care Facility at the University of Iowa under_a 12-h light-dark cycle with ad_libitum access to_food_and_water. All mice were_housed in the barrier facility until_time of craniectomy, and then remained_singly caged in a nonbarrier facility thereafter.

Fluid_percussion injury

Lateral FPI was performed as_previously described (Alder et al.,_2011). On_the day before injury, mice_underwent craniectomy. Animals_were anesthetized_with ketamine/xylazine (87_mg/kg ketamine and 12 mg/kg xylazine)_via intraperitoneal injection._The head was then mounted in_a_stereotaxic frame, and_a_midline_incision of_the scalp was_made_for reflection_of_the skin and exposure of underlying_skull._A 3-mm craniectomy was performed on the_left parietal skull bone_centered_between lambda and bregma_sutures and between lateral skull_edge and sagittal suture, using a_3.0-mm OD_handheld trephine_(University of Pennsylvania Machine Shop). A modified Luer-Lock hub was placed_surrounding the craniectomy site and secured_with cyanoacrylate glue. The_hub was_further_secured with methyl-methacrylate_dental_cement (Jet_Acrylic Liquid mixed with Perm Reline/Repair Resin)_surrounding the_bottom portion of the hub. The_hub was filled with_sterile_0.9% saline, and a sterile IV_cap was placed to prevent exposure_of the underlying dura to_the_environment_until time of FPI. The_next day, mice underwent FPI. Mice_received 3% inhaled_isoflurane in an induction chamber before being_transferred_to a nose cone where IV_cap_was removed and any air bubbles_in_the_hub were also removed. Once_deeply anesthetized, mice were connected to_the FPI device via 20-inch IV tubing and placed_on their right_side. A pendulum was released_from_an_angle of 11–12 degrees against the fluid reservoir, generating a_brief fluid_pulse against the_exposed dura