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Glutamate Transporters (GLT1, GLAST, EAAC1, EAAT4, and EAAT5), GTRAP41, GTRAP48, inhibitory protein factor (IPF), and Alpha-Fodrin Antibodies

A distinct step in inter cellular communication involves termination of synaptic transmission via the removal of neurotransmitters by specialized transporters. The regulated exocytotoic release of neurotransmitters in response to neural activity requires storage within intracellular vesicles. In the nervous system, these vesicles are the synaptic vesicles that are derived from the endosomal compartment, whereas in endocrine cells larger secretory granules, such as the chromaffin granules of adrenal medulla, are derived from the trans golgi networks. Glutamate is the main excitatory neurotransmitter in the brain. To date five glutamate Transporters have been cloned: GLAST (EAAT1), GLT1 (EAAT2), EAAC1 (EAAT3), EAAT4, and EAAT5. These transporters are believed to be critical in reducing potentially toxic extracellular concentration of glutamate by rapid uptake into nerve terminals and glial cells. Abnormalities in glutamate transport have been observed in amyotrophic lateral sclerosis. Glutamate transporters (525-573 AA) display about 55% homology and are predicted to contain up to 6-10 transmembrane domains. Immunolocalization studies indicate that GLT1 is localized in astroglial cells throughout the brain and spinal cord. EAAC1 is specific for certain neurons and purkinje cells, and specifically enriched in cortex, hippocampus, and caudate-putamen and confined to presynaptic and postsynaptic elements. GLAST has been observed in both neuron and astroglia. It is most abundant in Bergmann glia, cortex, hippocampus and cerebellum. EAAT4 has properties of ligand gated Cl-channel. It is localized mainly in cerebellar Purkinje cells in rat and human CNS. EAAT5 has only been cloned from human. It is primarily expressed in retina.

Most recently two proteins, designated GTRAP41 and GTRAP48 (for glutamate transporter EAAT4 associated protein) that specifically interact with the intracellular carboxy-terminal domain of EAAT4 have been cloned and characterized. The proteins are proposed to modulate glutamate transport activity. GTRAP41 is 2388 aa protein (~270 kDa). It has seventeen 16-amino acid spectrin repeats, 2 alpha-actinin domains, and a pleckstrin homology domain. GTRAP48 is 1527 aa (~170 kDa). GTRAP48 possesses a PDZ domain, a regulatory G protein-signaling sequence, tandem dbl homology and pleckstrin homology domains characteristic of guanine nucleotide exchange factors for the Rho family of G proteins, and 2 proline-rich sequences. brain tissue. GTRAP41 and GTRAp48 are mainly expressed in the brain, and lower levels were also detected in liver and kidney. EAAT4, GTRAP41, and GTRAP48 are all immunolocalized in cerebellar Purkinje cell soma and dendrites, with little axonal staining. GTRAP48 was found to activate Rho. Expression of GTRAP48 induced the reorganization of the actin cytoskeleton. When GTRAP41 and GTRAP48 are coexpressed, GTRAP41 colocalized with actin in structures that resemble actin-stress fibers, a typical Rho-dependent effect. Overexpression of GTRAP41 and GTRAP48 enhanced glutamate uptake. GTRAP41 and GTRAP48 may therefore enhance glutamate transport either through an increase in the catalytic rate of the transporter or through an increase in cell-surface availability or by stabilizing EAAT4 at the membrane.

Synaptic vesicles in the nerve terminals play a critical role in neurotransmission. Glutamergic neurotransmission occurs through an exocytotic process involving the interaction of glutamate containing synaptic vesicles with the plasma membranes of the presynaptic ending. An electro-chemical proton gradient generated by a V-type H+-ATPase (vacuolar-type proton-translocating ATPase) in the synaptic vesicles membrane provides the driving force for glutamate uptake. Recently a protein, termed inhibitory protein factor (IPF), has been isolated from brain cytosol that inhibits glutamate and GABA uptake into synaptic vesicles (IC50 ~25 nM). IPF does not inhibit ATP-independent uptake, norepinephrine uptake into chromaffin vesicles, and Na-dependent glutamate uptake into synaptosomes. IPF refers to a three distinct proteins with ~mol wt of 138kDa (IPF-alpha), 135 kDa (IPF-beta), and 132 kDa (IPF-gamma). IPF-a is derived from a ubiquitous, non-erythroid brain spectrin called alpha-Fodrin, a well-characterized protein previously implicated in such diverse activities as exocytosis/endocytosis, apoptosis, and NMDA-receptor activation. However, a-Fodrin itself has no effect on glutamate uptake. The N-terminal 1-20 aa of IPF-a, IPF-b, and IPF-g are identical with 26-45 aa of a-Fodrin (mol wt ~240 kDa). Therefore, it appears that some identified protease(s) may generate IPF-a from a-Fodrin.

 

M= Mouse; R=Rat; H=Human; Ha=Hamster; Rb=Rabbit; B=Bovine; CT= near C-terminus; NT=near N-terminus; Internal=Middle of protein. *
** Expected antibody crossreactivity information is mostly based upon high (>70%) sequence conservation of antigenic/control peptides in various species. When antibody crossreactivity has actually been experimentally confirmed in various species, it will be mentioned in the appropriate data sheets.
 

"Neat Antisera" are the unpurified antiserum and it is suitable for ELISA and Western.
"Affinity pure" IgG may be more suitable for immunohistochemical (IHC) applications and to reduce background in most immunological applications including ELISA and Western.
"Control peptides" can not be used for Western as they are very short peptides. They are intended for ELISA or antibody blocking studies to establish antibody specificity.
 

Citations of ADI's antibody for various glutamate transporters:

GLT1 Rodriguez-Kern A 2003 Neurochemistry International 43, 363-370 Beta-amyloid and brain-derived neurotrophic factor, BDNF, up-regulate the expression of glutamate transporter GLT-1/EAAT2 via different signaling pathways utilizing transcription factor NF-B, WB glt1 from SC/sigma too rat cortical astrocytes.

GLT1 Voutsinos-Porche 2003 Neuron, Volume 37, Issue 2, 23 January 2003, Pages 275-286 Glial Glutamate Transporters Mediate a Functional Metabolic Crosstalk between Neurons and Astrocytes in the Mouse Developing Cortex WB IHC.

GLT1 Levenson J 2002 Nature Neuroscience5, 155 - 161 Long-term potentiation and contextual fear conditioning increase neuronal glutamate uptake WB.

GLT1 Ueda Y 2002 Mol. brian. Res. 104, 120-126 Expression of glutamate transporters and ionotropic glutamate receptors in GLAST knockout mice WB, glast knockout mice.

GLT1 Ueda Y et al 2001 J. Neurochem. 2001 76: 892-900 Collapse of extracellular glutamate regulation during epileptogenesis: down-regulation and functional failure of glutamate transporter function in rats with chronic seizures induced by kainic acid WB.

GLT1 Ueda Y 2000 Exp. Brain Res. 133, 334-340 Molecular Regulation of glutamate and GABA transporters proteins by valproic acid in rat hippocampus during epileptogenesis WB.

 

GLT1 Ueda Y 2000 Epilepsy Res. 39, 201-209 Sequential change in glutamate transporter protein levels during Fe-induced epileptogenesis WB.

GLT1 Samuelsson C 2000 Neurosci. lett. 289, 185-188 Decreased Cortical Levels of astrocytic glutamate transport protein GLT-1 in a rat modle f posttraumatic epilepsy.

GLT1 Palos, Teresa P. 1999 J. Neurochemistry 73, 1012 Wnt Signaling Induces GLT-1 Expression in Rat C6 Glioma Cells WB, IHC, pc6 cells rat.

GLT1 Li, Shuxin 1999 J. Neurosci. 99, RC16, 1-9 Novel Injury Mechanism in Anoxia and Trauma of Spinal Cord White Matter: Glutamate Release via Reverse Na+-dependent Glutamate Transport IHC, other EAAc1 from toehrs 4% paraformaldehyde.

GLAST Voutsinos-Porche 2003 Neuron, Volume 37, Issue 2, 23 January 2003, Pages 275-286 Glial Glutamate Transporters Mediate a Functional Metabolic Crosstalk between Neurons and Astrocytes in the Mouse Developing Cortex WB IHC.

GLAST Rodriguez-Kern A 2003 Neurochemistry International 43, 363-370 Beta-amyloid and brain-derived neurotrophic factor, BDNF, up-regulate the expression of glutamate transporter GLT-1/EAAT2 via different signaling pathways utilizing transcription factor NF-B, WB glt1 from SC/sigma too rat cortical astrocytes.

GLAST Levenson J 2002 Nature Neuroscience5, 155 - 161 Long-term potentiation and contextual fear conditioning increase neuronal glutamate uptake WB.

GLAST Ueda Y 2002 Mol. brian. Res. 104, 120-126 Expression of glutamate transporters and ionotropic glutamate receptors in GLAST knockout mice WB, glast knockout mice.

GLAST Deitch JS et al 2002 J Neurol. Sci. 193, 117-126 GLT-1 glutamate transporter levels are unchanged in mice expressing G93A human mutant SOD1 WB, IHC, glt-1 from aff bio wb mouse brain, discuss glt-1 size and polymer, new buffer.

GLAST Ueda Y et al 2001 J. Neurochem. 2001 76: 892-900 Collapse of extracellular glutamate regulation during epileptogenesis: down-regulation and functional failure of glutamate transporter function in rats with chronic seizures induced by kainic acid WB.

GLAST Ueda Y 2000 Exp. Brain Res. 133, 334-340 Molecular Regulation of glutamate and GABA transporters proteins by valproic acid in rat hippocampus during epileptogenesis WB.

GLAST Ueda Y 2000 Epilepsy Res. 39, 201-209 Sequential change in glutamate transporter protein levels during Fe-induced epileptogenesis WB.

GLAST Samuelsson C 2000 Neurosci. lett. 289, 185-188 Decreased Cortical Levels of astrocytic glutamate transport protein GLT-1 in a rat modle f posttraumatic epilepsy.

GLAST Alexander, GM et al 2000 J. Neurochem. 74, 1666-1673 Elevated Cortical Extracellular Fluid Glutamate in Transgenic Mice Expressing Human Mutant (G93A) Cu/Zn Superoxide Dismutase WB, glt1 from affinity bio no difference; data not shown.

EAAC1 Voutsinos-Porche 2003 Neuron, Volume 37, Issue 2, 23 January 2003, Pages 275-286 Glial Glutamate Transporters Mediate a Functional Metabolic Crosstalk between Neurons and Astrocytes in the Mouse Developing Cortex WB IHC.

EAAC1 Rodriguez-Kern A 2003 Neurochemistry International 43, 363-370 Beta-amyloid and brain-derived neurotrophic factor, BDNF, up-regulate the expression of glutamate transporter GLT-1/EAAT2 via different signaling pathways utilizing transcription factor NF-B, WB glt1 from SC/sigma too rat cortical astrocytes.

EAAC1 Levenson J 2002 Nature Neuroscience5, 155 - 161 Long-term potentiation and contextual fear conditioning increase neuronal glutamate uptake WB.

EAAC1 Ueda Y 2002 Mol. brian. Res. 104, 120-126 Expression of glutamate transporters and ionotropic glutamate receptors in GLAST knockout mice WB, glast knockout mice.

EAAC1 Rome S et al 2002 J. Nutr. 132, 1009-1011 The Regionalization of PepT1, NBAT and EAAC1 Transporters in the Small Intestine of Rats Are Unchanged from Birth to Adulthood IHC pepT1, NBAT others rat intestine, paraffain sections.

EAAC1 Deitch JS et al 2002 J Neurol. Sc. 193, 117-126 GLT-1 glutamate transporter levels are unchanged in mice expressing G93A human mutant SOD1 WB, IHC, glt-1 from aff bio Formalin-fixed, paraffin-embedded tissue.

EAAC1 Ueda Y et al 2001 J. Neurochem. 2001 76: 892-900 Collapse of extracellular glutamate regulation during epileptogenesis: down-regulation and functional failure of glutamate transporter function in rats with chronic seizures induced by kainic acid WB.

EAAC1 Ueda Y 2000 Exp. Brain Res. 133, 334-340 Molecular Regulation of glutamate and GABA transporters proteins by valproic acid in rat hippocampus during epileptogenesis.

EAAC1 Ueda Y 2000 Epilepsy Res. 39, 201-209 Sequential change in glutamate transporter protein levels during Fe-induced epileptogenesis WB.

EAAC1 Alexander, GM et al 2000 J. Neurochem. 74, 1666-1673 Elevated Cortical Extracellular Fluid Glutamate in Transgenic Mice Expressing Human Mutant (G93A) Cu/Zn Superoxide Dismutase WB, glt1 from affinity bio no difference; data not shown.

EAAT4 Rodriguez-Kern A 2003 Neurochemistry International 43, 363-370 Beta-amyloid and brain-derived neurotrophic factor, BDNF, up-regulate the expression of glutamate transporter GLT-1/EAAT2 via different signaling pathways utilizing transcription factor NF-B, WB glt1 from SC/sigma too rat cortical astrocytes

EAAT4 Levenson J 2002 Nature Neuroscience5, 155 - 161 Long-term potentiation and contextual fear conditioning increase neuronal glutamate uptake WB.

EAAT4 Dunlop, J 1999 Brain Res. 839, 235-242 Properties of excitatory amino acid transport in the human U373 astrocytoma cell line.

 

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