The changing cellular localization of α-crystallin in the lens of the chicken embryo, studied by immunofluorescence

[摘要] Theappearanceanddistributionofα-crystallininchickenembryosandinanimalsafterhatchingwerestudiedwiththeindirectfluorescentantibodymethod.α-Crystallinwasisolatedbytwo-dimensionalpolyacrylamidegelelectrophoresis.Antiserawerepreparedagainstthepurifiedfractionandagainsttotalchickenlens.TheywerecontrolledfortheirspecificityinOuchterlonyplatesandimmunoelectrophoresis.Thefirstfluorescencewithtotallensantiserumwasobservedinthemorphologicallymostadvancedcellsofthelensplacodeof23-somiteembryos(50–53hoursofincubation).α-Crystallinantiseradidnotreactatthistime,indicatingthatanotherstructuralproteinwassynthesizedbeforeα-crystallinappeared.Withfurtherdevelopmentofthelensrudimentthefluorescencegraduallyspreadtomoreperipheralparts,andat3to3/12daystheentirelensreactedwithtotallensantiserum.Atthistimeα-crystallincouldfirstbedetectedinandaroundthenucleiofafewcentrallylocatedfibers.Graduallymoreperipheralfibersbecameinvolved,thentheequatorialzone,andfinallytheanteriorepithelium.Whilesomecellsreactedverystrongly,otherswereonlyweaklyfluorescentorevenremainednegative;thereresultedamosaiclikestainingpattern.At8daysofembryoniclifeallepithelialcellswerepositiveforα-crystallinandthereactioninthefibersbecameprogressivelyweaker.Intheadultlensthecenterofthelenswasnegative,theepithelialcellsandequatorialzone,withtheexceptionofafewcompletelynegativecells,werestronglypositiveandthesuperficialfibersshowedanintermediatereaction.Thefluorescencewasalwaysconfinedtothelens.Itisconcludedthatlensplacodeformationisindependentfromcrystallinsynthesis.Furthermore,theproductionofcrystallinsisrestrictedtothelensrudimentandlensanddoesnottakeplaceintheopticcupanditsderivatesorinotherectodermaltissues.α-Crystallinappearsrelativelylateindevelopmentandcannotbeconsideredasanessentialproteinforlensformation.

[学科分类] 生物科学（综合）