Irene Poyatos - Academia.edu (original) (raw)
Papers by Irene Poyatos
Neuroscience Letters, 1998
We have identified two alternative 5′ ends for the GLYT2 glycine transporter in rat brain DNA by ... more We have identified two alternative 5′ ends for the GLYT2 glycine transporter in rat brain DNA by using rapid amplification of cDNA ends (RACE) analysis. Study of the genomic DNA revealed that the isoform diversity is generated by alternative use of exons Ia or Ib, respectively. Upon translation, the mRNA corresponding to the novel isoform GLYT2b would yield a protein five amino acids longer than the previously characterized isoform GLYT2a. Both forms display similar regional distribution and kinetics characteristics. However, whereas GLYT2a is able to actively accumulate glycine into transfected COS cells, GLYT2b seems only to exchange (or release) glycine.
Molecular and Cellular Neuroscience, 2000
Asymmetrical distribution of Na ؉ -and Cl ؊ -dependent neurotransmitter transporters on the cell ... more Asymmetrical distribution of Na ؉ -and Cl ؊ -dependent neurotransmitter transporters on the cell surface of polarized cells seems to be a generalized feature in this gene family.
Glia, 1997
Two membrane-localized transporter proteins (GLYT1 and GLYT2) are responsible for removal of extr... more Two membrane-localized transporter proteins (GLYT1 and GLYT2) are responsible for removal of extracellular glycine in the mammalian CNS. Whereas GLYT1 seems to be expressed mainly in glial cells, GLYT2 is neuronal. The highest concentrations of both transporters are found in glycinergic areas of the nervous system. The expression of these proteins may be under regulatory control. We demonstrate here that GLYT1 is not expressed in pure glial cultures, but it is expressed by diverse types of glial cells in mixed neuronal/glial cultures. In these mixed cultures, the glial expression of GLYT1 is down-regulated after selective elimination of the neurons. The absence of expression in pure glial cultures and the observed reduction in the GLYT1 expression after neuronal loss support the existence of a regulatory cross-talk between neurons and glia to initiate and sustain the glial expression of GLYT1.
Molecular Brain Research, 1997
The glycine transporter GLYT2 is present in neurons of the spinal cord, the brain stem and the ce... more The glycine transporter GLYT2 is present in neurons of the spinal cord, the brain stem and the cerebellum. This localization is similar to that of glycine immunoreactivity, suggesting a causal relationship between GLYT2 expression and glycine distribution. In this report, we analyzed if such a relationship does exist by using neuronal cultures derived from embryonic spinal cord. GLYT2 was synthesized in a small subpopulation of neurons where it was targeted both to dendrites and to axons, being the axonal content higher than the dendritic one. At early stages in the development of cultured spinal neurons, the highest GLYT2 levels were found in the axonal growth cones. As the culture matured, immunoreactivity extended to the axonal shaft. Double-immunofluorescence experiments indicated a perfect co-localization of GLYT2 and glycine immunoreactivity in cultured neurons. Moreover, the concentration of glycine into neurons expressing GLYT2 was proportional to the concentration of the transporter. This observation was reproduced in GLYT2-transfected COS cells. These evidences indicate that the high content of glycine observed in some neurons in culture is indeed achieved by the concentrative task performed by GLYT2, and that GLYT2 can be used as a reliable marker for identification of glycine-enriched neurons. q 1997 Elsevier Science B.V.
Journal of Biological Chemistry, 2001
Glycine transporter GLYT2 is an axonal glycoprotein involved in the removal of glycine from the s... more Glycine transporter GLYT2 is an axonal glycoprotein involved in the removal of glycine from the synaptic cleft. To elucidate the role of the carbohydrate moiety on GLYT2 function, we analyzed the effect of the disruption of the putative N-glycosylation sites on the transport activity, intracellular traffic in COS cells, and asymmetrical distribution of this protein in polarized Madin-Darby canine kidney (MDCK) cells. Transport activity was reduced by 35-40% after enzymatic deglycosylation of the transporter reconstituted into liposomes.
Neuroscience Letters, 1998
We have identified two alternative 5′ ends for the GLYT2 glycine transporter in rat brain DNA by ... more We have identified two alternative 5′ ends for the GLYT2 glycine transporter in rat brain DNA by using rapid amplification of cDNA ends (RACE) analysis. Study of the genomic DNA revealed that the isoform diversity is generated by alternative use of exons Ia or Ib, respectively. Upon translation, the mRNA corresponding to the novel isoform GLYT2b would yield a protein five amino acids longer than the previously characterized isoform GLYT2a. Both forms display similar regional distribution and kinetics characteristics. However, whereas GLYT2a is able to actively accumulate glycine into transfected COS cells, GLYT2b seems only to exchange (or release) glycine.
Molecular and Cellular Neuroscience, 2000
Asymmetrical distribution of Na ؉ -and Cl ؊ -dependent neurotransmitter transporters on the cell ... more Asymmetrical distribution of Na ؉ -and Cl ؊ -dependent neurotransmitter transporters on the cell surface of polarized cells seems to be a generalized feature in this gene family.
Glia, 1997
Two membrane-localized transporter proteins (GLYT1 and GLYT2) are responsible for removal of extr... more Two membrane-localized transporter proteins (GLYT1 and GLYT2) are responsible for removal of extracellular glycine in the mammalian CNS. Whereas GLYT1 seems to be expressed mainly in glial cells, GLYT2 is neuronal. The highest concentrations of both transporters are found in glycinergic areas of the nervous system. The expression of these proteins may be under regulatory control. We demonstrate here that GLYT1 is not expressed in pure glial cultures, but it is expressed by diverse types of glial cells in mixed neuronal/glial cultures. In these mixed cultures, the glial expression of GLYT1 is down-regulated after selective elimination of the neurons. The absence of expression in pure glial cultures and the observed reduction in the GLYT1 expression after neuronal loss support the existence of a regulatory cross-talk between neurons and glia to initiate and sustain the glial expression of GLYT1.
Molecular Brain Research, 1997
The glycine transporter GLYT2 is present in neurons of the spinal cord, the brain stem and the ce... more The glycine transporter GLYT2 is present in neurons of the spinal cord, the brain stem and the cerebellum. This localization is similar to that of glycine immunoreactivity, suggesting a causal relationship between GLYT2 expression and glycine distribution. In this report, we analyzed if such a relationship does exist by using neuronal cultures derived from embryonic spinal cord. GLYT2 was synthesized in a small subpopulation of neurons where it was targeted both to dendrites and to axons, being the axonal content higher than the dendritic one. At early stages in the development of cultured spinal neurons, the highest GLYT2 levels were found in the axonal growth cones. As the culture matured, immunoreactivity extended to the axonal shaft. Double-immunofluorescence experiments indicated a perfect co-localization of GLYT2 and glycine immunoreactivity in cultured neurons. Moreover, the concentration of glycine into neurons expressing GLYT2 was proportional to the concentration of the transporter. This observation was reproduced in GLYT2-transfected COS cells. These evidences indicate that the high content of glycine observed in some neurons in culture is indeed achieved by the concentrative task performed by GLYT2, and that GLYT2 can be used as a reliable marker for identification of glycine-enriched neurons. q 1997 Elsevier Science B.V.
Journal of Biological Chemistry, 2001
Glycine transporter GLYT2 is an axonal glycoprotein involved in the removal of glycine from the s... more Glycine transporter GLYT2 is an axonal glycoprotein involved in the removal of glycine from the synaptic cleft. To elucidate the role of the carbohydrate moiety on GLYT2 function, we analyzed the effect of the disruption of the putative N-glycosylation sites on the transport activity, intracellular traffic in COS cells, and asymmetrical distribution of this protein in polarized Madin-Darby canine kidney (MDCK) cells. Transport activity was reduced by 35-40% after enzymatic deglycosylation of the transporter reconstituted into liposomes.