Age-related Changes in the Global DNA Methylation Profile of Oligodendrocyte Progenitor Cells Derived from Rat Spinal Cords

Demyelination of axons plays an important role in the pathology of many spinal cord diseases and injuries.Remyelination in demyelinated lesions is primarily performed by oligodendrocyte progenitor cells(OPCs),which generate oligodendrocytes in the developing and mature central nervous system.The efficiency of remyelination decreases with age.Many reports suggest that this decline in remyelination results from impaired OPC recruitment and differentiation during aging.Of the various molecular mechanisms involved in aging,changes in epigenetic modifications have received particular attention.Global DNA methylation is a major epigenetic modification that plays important roles in cellular senescence and organismal aging.Thus,we aimed to evaluate the dynamic changes in the global DNA methylation profiles of OPCs derived from rat spinal cords during the aging process.We separated and cultured OPCs from the spinal cords of neonatal,4-month-old,and 16-month-old rats and investigated the age-related alterations of genomic DNA methylation levels by using quantitative colorimetric analysis.To determine the potential cause of dynamic changes in global DNA methylation,we further analyzed the activity of DNA methyltransferases(DNMTs)and the expression of DNMT1,DNMT3a,DNMT3b,TET1,TET2,TET3,MBD2,and MeCP2 in the OPCs from each group.Our results showed the genomic DNA methylation level and the activity of DNMTs from OPCs derived from rat spinal cords decreased gradually during aging,and OPCs from 16-month-old rats were characterized by global hypomethylation.During OPC aging,the mRNA and protein expression levels of DNMT3a,DNMT3b,and MeCP2 were significantly elevated;those of DNMT1 were significantly down-regulated;and no significant changes were observed in those for TET1,TET2,TET3,or MBD2.Our results indicated that global DNA hypomethylation in aged OPCs is correlated with DNMT1 downregulation.Together,these data provide important evidence for partly elucidating the mechanism of age-related impaired OPC recruitment and differentiation and assist in the development of new treatments for promoting efficient remyelination.

Quantitative analysis by reversed-phase high-performance liquid chromatography and retinal neuroprotection after topical administration of moxonidine

AIM:To determine moxonidine in aqueous humor and iris-ciliary body by reversed-phase high performance liquid chromatography(RP-HPLC),and to evaluate the retinal neuroprotective effect after topical administration with moxonidine in a high intraocular pressure(IOP)model.METHODS:The eyes of albino rabbits were administered topically and ipsilaterally with 0.2%moxonidine.A RPHPLC method was employed for the identification and quantification of moxonidine between 2 and 480 min,which presented in the aqueous humor and iris-ciliary body.Flash electroretinography(F-ERG)amplitude and superoxide dismutase(SOD)level were measured between day 1 and day 15 after topical administration with moxonidine in a rabbit model of high IOP.Histological and ultrastructural observation underwent to analyze the changes of retinal morphology,the inner retinal layers(IRL)thickness,and retinal ganglion cell(RGC)counting.RESULTS:Moxonidine was detectable between 2 and 480 min after administration,and the peak concentration developed both in the two tissues at 30 min,0.51μg/m Lin aqueous humor and 1.03μg/g in iris-ciliary body.In comparison to control,F-ERG b-wave amplitude in moxonidine eyes were significantly differences between day 3 and day 15(P<0.01)in the high IOP model;SOD levels were significantly higher at all time-points(P<0.01)with a maximum level of 20.29 U/mgprot at day 15;and RGCs were significantly higher(P<0.05).CONCLUSION:Moxonidine is a viable neuroprotective agent with application to high IOP model.All layers of retina,including RGC layer,retinal nerve fiber layer and INL,are more preserved after moxonidine administration.SOD plays a neuroprotective role in ocular hypertension-mediated RGC death.