| Objective: The aim of this st?dy was to investigate the role of tubulin folding cofactor B(TBCB)in Fetal Alcohol Syndrome(FAS).The pathogenesis of FAS was st?died from the perspective of TBCB reg?lating microt?b?le biosynthesis and dynamic balance.Methods: A mo?se model of fetal alcohol syndrome was established,and the ne?rological symptoms of fetal alcohol syndrome were determined by behavioral test d?ring its development.Changes in the n?mber of ne?rons and dendritic spines and the length of fiber axons in the brain of P0 newborn mice were detected by Nissl and Golgi staining.The ?ltrastr?ct?ral changes in the cell body and fiber of ne?rons were detected by transmission electron microscopy.Imm?noflo?rescent(IF)and Western Blot(WB)detected the expression changes of ne?ron marker Neu N,dendritic marker Map2,axon marker Smi,tubulin alpha-t?blin and TBCB in the brain,and the expression changes of ne?ron,tubulin and TBCB were confirmed.Q?antitative Reverse Transcription PCR(qrt-pcr)was ?sed to detect the m RNA expression changes of TBCB.Res?lts: Compared with normal mice,FAS newborn mice had significantly lower birth weight and no significant difference in ne?rodevelopmental observation.D?ring its development,behavioral tests showed that the development of proprioception and motor coordination ability of FAS mice was significantly delayed,the ability to explore new things was red?ced,and the learning and memory ability was significantly red?ced.The n?mber of ne?rons in the brain of newborn mice was detected,and the staining res?lts of Nissl and Golgi showed no significant difference between the n?mber of ne?rons in FAS mice and the control gro?p.The morphology and str?ct?re of the cell body of ne?rons in the brain of newborn mice were detected,and the morphological str?ct?re of the cell body and n?cle?s,as well as the n?mber and morphological str?ct?re of organelles of most of the ne?rons in FAS newborn mice were fo?nd to be normal by transmission electron microscope.IF and WB showed that the expression of axon and dendrite markers Smi and Map2 in the brain of FAS mice were significantly decreased.The res?lts of electron microscopy showed that in the longit?dinal section of cortical nerve fibers in FAS newborn mice,the thin and long t?b?lar microt?b?les in parallel arrangement almost disappeared,and ribosomes and mitochondria edema and decrease,b?t still visible.The hollow circ?lar str?ct?re of microt?b?les disappeared and presented a flocc?lent irreg?lar str?ct?re.After the detection of TBCB and microt?b?les in the brain of newborn mice,IF showed that the expression of these two proteins was significantly decreased in the motor region,amygdala and thalam?s of the brain of newborn FAS mice,WB showed that the expression of these two proteins was significantly decreased in the whole brain of newborn FAS mice,and qrt-pcr showed the down-reg?lation of TBCB m RNA in the whole brain of newborn FAS mice.TBCB and alpha-tubulin were co-expressed along microt?b?les,b?t the co-expression area and n?mber were significantly red?ced.brain,amygdala and thalam?s of FAS mice were significantly decreased.Concl?sion: The down-reg?lation of TBCB expression in the brain of FAS neonatal mice leads to serio?s damage of microt?b?le morphology and str?ct?re,which leads to ne?rofibropathy,and ?ltimately leads to ne?rodevelopmental disorders in FAS mice,showing delayed development of learning,memory,perception and sensation.Objective: After tra?matic brain inj?ry(TBI)in rats and astrocyte inj?ry,the expression patterns of aq?aporin-4(AQP4)and dystroglycan(DG)were st?died to explore whether the expression changes of AQP4 were related to DG,and whether the both expression changes were reg?lated by ERK signaling pathway.Methods: The rats were randomly divided into TBI gro?p and control gro?p,and the TBI model was established by free fall method.Tiss?es were taken from each gro?p at 6 h,12 h,24 h and 48 h after s?rgery.The level of blood brain barrier(BBB)inj?ry was detected by Evans Bl?e(EB)method,the protein expression changes of AQP4,? and ?-DG in the control gro?p and the TBI gro?p were detected by IF and WB,and the m RNA content changes of the three gro?ps were detected byq RT-PCR.In order to eliminate the interference of secondary factors of TBI and clarify the relationship between AQP4 and DG expression and its reg?latory mechanism in astrocytes after TBI.Primary astrocytes were c?lt?red in newborn SD rats and randomly divided into gro?ps to establish the scratch and false scratch control gro?p models.The protein and m RNA expression changes of AQP4,? and ?-DG were detected by IF,WB and RT-PCR in the scratch gro?p and the control gro?p at 6 h and 24 h after the scratch.WB detects changes in ERK signaling pathway.In order to determine whether ERK signaling pathway is involved in the reg?lation of AQP4 and DG expression after TBI,astrocytes were randomly divided into control gro?p,inhibitor gro?p and agonist gro?p.DMSO,?0126 and TPA were added to pretreat for 1 h respectively for scratching,and the cells were collected 6 h after scratching.The protein and m RNA expression of AQP4 and AQP4,? and ?-dg were detected by IF,WB and RT-PCR.Res?lts: In each time period after TBI,the expression changes of AQP4,? and ?-dg are basically the same,b?t d?e to the interference of the secondary inflammatory response and other factors after TBI,the expression changes of DG and AQP4 are indeed inconsistent.However,in the p?rified astrocytes,the changes of DG and AQP4 expression after scratch were completely consistent with the changes of P-ERK.After interfering with the ERK signal pathway,the expression of DG and AQP4 will change correspondingly.Concl?sion: After TBI,DG participates in the anchoring of AQP4 in astrocytes,b?t it is not the excl?sive anchoring protein of the polar expression of AQP4.ERK signaling pathway is activated in early stage after mechanical inj?ry,and is involved in the reg?lation of DG and AQP4 expression. |
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