Anti-SLC1A3 monoclonal antibody, clone FQS23797

Active transporters play a central role in the uptake and efflux of glutamate by cells and organelles. We aimed to assess the effects of crowding stress on the efficiency of active glutamate and glutamine transport in fish. We cloned intestinal-related transporters and investigated their transcriptional changes in grass carpCtenopharyngodon idellusexposed to different stocking densities. The full-length cDNA sequences ofslc1a1,slc1a3andslc7a6cloned by rapid amplification of cDNA ends were 2,869, 2,508 and 2,141 base pairs respectively. Based on homology modelling, we derived three-dimensional models of SLC1A1, SLC1A3, SLC7A6 and SLC7A7 and noted their transmembrane domains. The expression profiles ofslc1a1,slc1a3andslc7a7demonstrated tissue specificity, with section-dependent expression patterns along the intestine. Moreover, segmental mRNA expression patterns ofslc1a3,slc7a6andslc7a7were affected by stocking density. Crowding stress notably altered the segment-specific mRNA expression ofslc7a6. The results of this study showed that increased stocking density increased mRNA expression levels of glutamine transporters (slc7a6andslc7a7) but decreased mRNA expression of the glutamate transporter (slc1a3). These results furthered our understanding of the relationship between crowding stress and intestinal glutamate/glutamine transport in fish.

Purpose: This study investigated changes in the transcript levels of genes related to glutamate neurotransmission and transport as diabetes progresses in the Long-Evans rat retina. Transcript levels of vascular endothelial growth factor (VEGF), erythropoietin, and insulin-like growth factor binding protein 3 (IGFBP3) were also measured due to their protective effects on the retinal vasculature and neurons. Methods: Diabetes was induced in Long-Evans rats with a single intraperitoneal (IP) injection of streptozotocin (STZ; 65 mg/kg) in sodium citrate buffer. Rats with blood glucose >300 mg/dl were deemed diabetic. Age-matched controls received a single IP injection of sodium citrate buffer only. The retinas were dissected at 4 and 12 weeks after induction of diabetes, and mRNA and protein were extracted from the left and right retinas of each rat, respectively. Gene expression was analyzed using quantitative real-time reverse-transcription PCR. Enzyme-linked immunosorbent assay was used to quantify the concentration of VEGF protein in each retina. Statistical significance was determined using 2x2 analysis of variance followed by post-hoc analysis using Fisher's protected least squares difference. Results: Transcript levels of two ionotropic glutamate receptor subunits and one glutamate transporter increased after 4 weeks of diabetes. In contrast, 12 weeks of diabetes decreased the transcript levels of several genes, including two glutamate transporters, four out of five N-methyl-D-aspartate (NMDA) receptor subunits, and all five kainate receptor subunits. Diabetes had a greater effect on gene expression of NMDA and kainate receptor subunits than on the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor subunits, for which only GRIA4 significantly decreased after 12 weeks. VEGF protein levels were significantly increased in 4-week diabetic rats compared to age-matched control rats whereas the increase was not significant after 12 weeks. Transcript levels of VEGF and VEGF receptors were unchanged with diabetes. Erythropoietin and IGFBP3 mRNA levels significantly increased at both time points, and IGFBP2 mRNA levels increased after 12 weeks. Conclusions: Diabetes caused significant changes in the transcriptional expression of genes related to ionotropic glutamate neurotransmission, especially after 12 weeks. Most genes with decreased transcript levels after 12 weeks were expressed by retinal ganglion cells, which include glutamate transporters and ionotropic glutamate receptors. Two genes expressed by retinal ganglion cells but unrelated to glutamate neurotransmission, gamma-synuclein (SNCG) and adenosine A1 receptor (ADORA1), also had decreased mRNA expression after 12 weeks. These findings may indicate ganglion cells were lost as diabetes progressed in the retina. Decreased expression of the glutamate transporter SLC1A3 would lead to decreased removal of glutamate from the extracellular space, suggesting that diabetes impairs this function of Muller cells. These findings suggest that ganglion cells were lost due to glutamate excitotoxicity. The changes at 12 weeks occurred without significant changes in retinal VEGF protein or mRNA, although higher VEGF protein levels at 4 weeks may be an early protective response. Increased transcript levels of erythropoietin and IGFBP3 may also be a protective response.