The presence of ths 47.5 kDa band prompted us to nvestgate the expressoof GAtranscrpts spnal cord of Ganex1,ex1 mce.RT PCR analyses have been performed wth prmers to amplfy the regobetweeexons 1 and two at the same time because the regobetweeexons 2 and seven.Usng RT PCR prmers for exons 1 to 2, no band was detected CNS RNA samples from Ganex1,ex1, as anticipated from exo1 deleton.even so, RT PCR wth prmers for exons two and 7elded a band spnal cord samples from Ganex1,ex1 mce but not other tssues this kind of as braand lver.Ths transcrpt spans Gaexo2 to 11 and mssng exo1.As a result, the RT PCR effects confrm the lack of total lenght GamRNA Ganex1,ex1 mce however they help the exstence of shorter ggaxonspeces the spnal cord.Ths short ggaxonvarant wl be known as sGg.Alterations F protelevels the nervous procedure well establshed that ggaxondeprved tssues from GApatents current characterstc accumulatons of Fs.
We therefore examned no matter if the absence of exo1 Ganex1,ex1 mce resulted abnormal levels of Fs and of other cytoskeletal elements.Westerblot analyses of complete proteextracts at 3 months of age from your bran, cerebellum and spnal cord uncovered modest ncreases more hints protelevels of F protens ncludng NF L, NF M, NFh and nternexn.A two fold ncrease perpherand vmentlevels have been also observed the spnal cord of Ganex1,ex1 mce whecompared to standard lttermates.The analyss of scatc nerve sectons unveiled enhanced NF protelevels along the nerve.The 3 subunts seem to be to be even more abundant the nerve proxmal regon.The ncreased levels of neuronal F protens Ganex1,ex1 was not on account of ncreased mRNA expresson.
RT PCR analyss for NF L, NF M and NFh showed no dfferences of transcrpt amounts the bran, cerebellum or spnal cord pan JAK inhibitor of Ganex1,ex1 mce and lttermate controls.To determne f the varatons F protelevels have been frequent durng agng, dot blot mmunodetectoanalyses were

performed for NF subunts, nternexand vmentat dfferent ages.The results confrmed a deregulatoof all F protelevels early as 3 months of age that was stl existing at 24 months of age.partcular, NFh and NF L amounts were ncreased uto two folds whatsoever ages the bran, cerebellum and spnal cord samples of Ganex1,ex1mce.NF M levels remaned unchanged at six and twelve months of age cerebellum and spnal cord samples of Ganex1,ex1 mce.The nternexlevels have been ncreased by uto three.eight fold the cerebellum of Ganex1,ex1 mce at 3 months of age as in contrast to controls.the braas properly as the ventral a part of the lumbar spnal cord, the mmunostanng for NF L was more powerful but no nclusons have been detected.By far the most notable alterations came from your mmunodetectoof NFh and nternexn.the cerebral cortex these two protens formed accumulatons the neuronal cell bodes of Ganex1,ex1 mce.These F accumulatons appear as early as a single month of age and are stl present at twelve months of age.