Developmental expression of a synaptic vesicle-specific protein in the rat retina☆

摘要

Inordertoexaminetheappearanceofsynapticvesiclesandtocorrelateitwiththeformationofthesynapticlayers,wehavedeterminedthestainingpatternofamurinemonoclonalantibody(SV48)toasynapticvesicle-associatedproteinindevelopingratretina.Theantigenwasdetectedbytheindirectimmunofluorescencetechniqueusingcryostatsectionsofparaformaldehyde-fixedretinas.Intheadultretina,theantibodystainedboththeouterplexiform(OPL)andtheinnerplexiformlayers(IPL).Thenuclearlayersandthenervefiberlayer(NFL)weredevoidofanystaining.Inprenatalandearlypostnatal(P)retinas,theantibodystainedtwobandswhichcorrespondedtotherespectivelocationsoftheNFLandIPL.StainingintheNFLincreaseduntilP-4andbegantodeclinesubsequently,andbyP-8littlestainingwasleftinthislayer.Incontrast,intheIPL,theintensityofstainingincreasedgraduallyandleveledoffbyP-10.Intheouterretina,abandoffluorescencecorrespondingtotheOPLwasfirstobservedatP-5andincreasedinintensityuptoP-10.ImmunoblottingstudiesshowedthatthemajorimmunoreactivematerialfromadultandembryonicretinashadaMra??65,000a€“67,000.Asexpectedfromitsdevelopmentalpattern,allbandsappearedinitiallyinthecentralretinaandsubsequentlyintheperipheralretina.Ourresultsshowthatthesynapticvesicle-proteinispresentinthenervefiberlayerbeforesynaptogenesisinthecentralnervoussystem.Subsequently,theproteinislostfromtheNFL,possiblyasaconsequenceofsynapseformation.