Dataset Information

LC-MS/MS analysis of proteins interacting with EAAT3

ABSTRACT: The excitatory amino acid transporter-3 (EAAT3) mediates the reuptake of glutamate from the synaptic cleft to terminate synaptic transmission at glutamatergic synapses. The plasma membrane expression of EAAT3 is regulated by the cycle of endocytosis and exocytosis.In this project we aimed to identify interacting proteins of EAAT3 involved in the trafficking of EAAT3.

INSTRUMENT(S): Bruker Daltonics micrOTOF series

ORGANISM(S): Rattus Norvegicus (rat) Homo Sapiens (human) Cloning Vector Pegfp-c1

TISSUE(S): Brain, Cell Culture

SUBMITTER: Jae-Won Yang

LAB HEAD: Jae-Won Yang

PROVIDER: PXD024524 | Pride | 2021-04-14

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	10_EAAT3_RBL_box_15kD.baf	Other
	10_EAAT3_RBL_box_15kD_F008982.mzid.gz	Mzid
	11_GFP_RBL_arrow_corresp_250kD.baf	Other
	11_GFP_RBL_arrow_corresp_250kD_F008979.mzid.gz	Mzid
	12_GFP_RBL_bracket_corresp_130kD.baf	Other

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Project description:Fast-spiking (FS) interneurons are important elements of neocortical circuitry that constitute the primary source of synaptic inhibition in adult cortex and impart temporal organization on ongoing cortical activity. The highly specialized intrinsic membrane and firing properties that allow cortical FS interneurons to perform these functions are attributable to equally specialized gene expression, which is ultimately coordinated by cell-type-specific transcriptional regulation. Although embryonic transcriptional events govern the initial steps of cell-type specification in most cortical interneurons, including FS cells, the electrophysiological properties that distinguish adult cortical cell types emerge relatively late in postnatal development, and the transcriptional events that drive this maturational process are not known. To address this, we used mouse whole-genome microarrays and whole-cell patch clamp to characterize the transcriptional and electrophysiological maturation of cortical FS interneurons between postnatal day 7 (P7) and P40. We found that the intrinsic and synaptic physiology of FS cells undergoes profound regulation over the first 4 postnatal weeks and that these changes are correlated with primarily monotonic but bidirectional transcriptional regulation of thousands of genes belonging to multiple functional classes. Using our microarray screen as a guide, we discovered that upregulation of two-pore K leak channels between P10 and P25 contributes to one of the major differences between the intrinsic membrane properties of immature and adult FS cells and found a number of other candidate genes that likely confer cell-type specificity on mature FS cells. Experiment Overall Design: We characterized the transcriptional maturation of genetically labeled FS interneurons in mouse S1 cortex using whole-genome microarrays.mRNA was harvested from manually sorted GFPexpressing neurons dissociated from acutely prepared brain slices at a range of developmental time points (P7, P10, P15, P25, and P40) and subsequently reverse transcribed, amplified, labeled, and hybridized to Affymetrix 430 2.0 whole-genome microarrays (three microarrays from three different animals per condition). We used the G42 transgenic mice described by Chattopadhyaya et al. (2004) for all experiments.

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LC-MS/MS analysis of proteins interacting with EAAT3

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