A role for nuclear factor I in the intrinsic control of cerebellar granule neuron gene expression.
Nervous system formation requires the elaboration of a complex series of differentiation events in both a spatially and maturation-regulated manner. A fundamental question is how neuronal subtype specification and developmental gene expression are controlled within maturing neurons. The alpha6 subunit of the gamma-aminobutyric acid type A (GABA(A)) receptor (GABRA6) is preferentially expressed in cerebellar granule neurons and is part of an intrinsic program directing their differentiation. We have employed a lentiviral approach to examine the transcriptional mechanisms controlling neuronal subtype-selective expression of this gene. These studies demonstrated that nuclear factor I (NFI) proteins are required for both transgenic GABRA6 promoter activity as well as endogenous expression of this gene in cerebellar granule neurons. Chromatin immunoprecipitation also showed that NFI proteins are bound to the GABRA6 promoter in these cells in vivo. Furthermore, analyses of gene knockout mice revealed that Nfia is specifically required for normal expression of the GABRA6 gene in cerebellar granule neurons. NFI expression and DNA binding activity are highly enriched in granule neurons, implicating this transcription factor family in the neuronal subtype-selective expression of the GABRA6 gene. These studies define a new role for NFI proteins as neuronal subtype-enriched transcriptional regulators that participate in an intrinsic transcriptional program directing the differentiation of cerebellar granule neurons.
University of Massachusetts Medical School, Department of Molecular and Cellular Physiology, 55 Lake Ave N., Worcester, MA 01655, USA.
Wei
W
Wang
Rachel E
RE
Stock
Richard M
RM
Gronostajski
Yong Wee
YW
Wong
Melitta
M
Schachner
Daniel L
DL
Kilpatrick
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015153:Blotting, Western; D022762:CCAAT-Enhancer-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation; D002460:Cell Line; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism*; D050485:GABA Plasma Membrane Transport Proteins; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D016086:Lentivirus / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D051839:NFI Transcription Factors; D009474:Neurons / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011963:Receptors, GABA-A / Q000737:chemistry / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic; D019076:Transgenes
D022762:CCAAT-Enhancer-Binding Proteins; D002843:Chromatin; D050485:GABA Plasma Membrane Transport Proteins; C546171:GABRA6 protein, human; C546172:Gabra6 protein, mouse; D026901:Membrane Transport Proteins; D051839:NFI Transcription Factors; C497592:NFIA protein, human; C497646:Nfia protein, mouse; D011963:Receptors, GABA-A; D014157:Transcription Factors; D008156:Luciferases