Anmal Studes Female Sprague Dawley rats weghng 225 250 g werehoused a single to four per cage approxmately seven days pror for the begnnng of your review, wth zero cost entry to food and water and mantaned oa twelve,12, lght,dark schedule at 21 C and 60%humdty.Allhousng condtons and expermental procedures have been carred out accordance wth accredited nsttutonal anmal care and use protocols.Under soflurane anesthesa a C6 eight vertebral lamnectomy was carried out, exposng the C7 spnal cord segment wthout openng of the dura, and 160 Kdyne blunt force trauma was appled asymmetrcally on the rght dorsum of C7 spnal cord usng a one.5 mm dameter probe controlled by a computerzed mpactor devce.Onehour after njury 1 ul contanng 41010 plaque formng unts of ether vEPO or vC was njected over the leson, and aaddtonal 1 ul njected under the lesoorght sde in the spnal cord at a charge of 0.5 ul mn.Sham anmals underwent precisely the same spnal cord publicity but no njury or njectowas carried out.For the behavoral analyss, there have been 8 anmals just about every SC grouand 4 sham controls.
At the conclusoof the behavoral experments, anmals were perfused via theheart wth one hundred ml of phosphate buffered salne followed by 500 ml of 4% paraformaldehyde PBS.The spnal cord was eliminated, postfxed at 4 C overnght, and cryoprotected wth 30% sucrose PBS.For proteanalyss, 2 and 8 weeks right after selleck njury anmals have been perfused by theheart wth one hundred ml of PBS and spnal cord removed and quckly frozedry ce.Behavoral assessment Forelmb motor functowas evaluated usng the followng tests, Opefeld testng The opefeld envronment conssted of the crcular plexglas enclosure 95 cm dameter, forty cmhgh wth aant skd floor.Rats had been tested pars a shadowy lght to motivate locomotor actvty.Anmals that remaned selleck chemicals statonary for any perod longer tha20 sec have been pcked uand placed with the center within the opefeld arena to rentate locomoton.All rats were scored more than a tme wndow of the maxmum duratoof 4mn, whilst the observatoperod was sometmes extended to make absolutely sure that the anmals abtes have been not underestmated.
Lmb use asymmetry test Forelmb use durng exploratve actvty was analyzed by puttng rats a transparent cylnder for five mor 20 touchs durng the tral.A mrror was placed behnd the cylnder at aangle to enable the rater to record forelmb movements whethe

anmal was turned away from the rater.The cylndrcal shape encourages vertcal exploratoof the walls wth the forelmbs as well as landng actvty.Measurement of forelmb placng Forelmb placng asymmetry was scored usng the vbrssae elcted forelmb placng test.Anmals wereheld by ther torsos allowng forelmbs tohang free.Wheholdng the anmal, the expermenter made gentle uand dowmovements space pror to place testng, whch factated muscle relaxatoand elmnated any strugglng movements.