The expression of methyl CpG binding factor MeCP2 correlates with cellular differentiation in the developing rat brain and in cultured cells.
Mutations in the MeCP2 gene cause Rett syndrome, a neurologic condition affecting primarily young girls. To gain insight into the normal function of MeCP2, we examined its temporal and spatial expression patterns, and immunoreactive prevalence, during late embryonic and perinatal brain development. MeCP2 mRNA was detected in most regions of the developing rat brain by the late embryonic stage. Regions displaying the strongest mRNA expression include the hippocampus, cortex, and cerebellum, and moderate expression was observed in most other brain regions. At the protein level, MeCP2 was strongly expressed in adult forebrain neurons, but was not detected in astrocytes. The nonubiquitous expression of MeCP2 was also observed in the embryonic cortex, as about one-third of acutely dissociated embryonic day 14 neuroepithelial cells failed to stain with MeCP2. To test whether MeCP2 expression correlates with neuronal differentiation, colocalization of MeCP2 expression with either the precursor cell marker nestin or the young neuronal marker beta-III tubulin was examined in the same acutely dissociated cortical cells. Although strong MeCP2 expression was detected in approximately 75% of beta-III tubulin-positive cells, only about 25% of nestin-positive precursor cells were MeCP2 positive. Further support for a correlation of MeCP2 expression with cell differentiation was observed in culture, where Western blot analysis during the in vitro differentiation of PC12, NG108-15, and SH-SY5Y cells revealed that MeCP2 levels increased as the cells acquired a more differentiated phenotype. This increase was associated with differentiation, as MeCP2 expression levels did not vary within different phases of the cell cycle. Taken together, these data support a role for MeCP2 in the establishment and/or maintenance of neuronal maturity.
Jung
Benjamin P
BP
Division of Cellular and Molecular Biology, Mac 11-412, 399 Bathurst Street, Toronto Western Research Institute, University Health Network, 399 Bathurst Street, Toronto, Ontario M5T 2S8, Canada.
Jugloff
Denis G M
DG
Zhang
Guangming
G
Logan
Richard
R
Brown
Stephanie
S
Eubanks
James H
JH
D000818:Animals; D001921:Brain / Q000033:anatomy & histology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line / Q000378:metabolism; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone; D016213:Cyclins / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005314:Embryonic and Fetal Development / Q000235:genetics / Q000502:physiology*; D005260:Female; D005333:Fetus; D005786:Gene Expression Regulation; D015151:Immunoblotting; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D051783:Methyl-CpG-Binding Protein 2; D009474:Neurons / Q000378:metabolism; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012097:Repressor Proteins; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D016213:Cyclins; D004268:DNA-Binding Proteins; C497394:Mecp2 protein, rat; D051783:Methyl-CpG-Binding Protein 2; D012333:RNA, Messenger; D012097:Repressor Proteins; D014404:Tubulin