Production of specific antibodies against GABA transporter subtypes (GAT1, GAT2, GAT3) and their application to immunocytochemistry

Ikegaki, Natsu; Saito, Naoaki; Hashima, Makoto; Tanaka, Chikako

doi:10.1016/0169-328x(94)90072-8

Cited by 123 publications

(97 citation statements)

References 15 publications

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“…The staining patterns of GABA, glycine, VGAT, GAT1, GAT3, GLYT1 and GLYT2 in the brain were similar to those described previously (Ottersen et al, 1988;Chaudhry et al, 1998;Ikegaki et al, 1994;Itouji et al, 1996;Zafra et al, 1995). In particular, in spite of the lack of GLYT1 and GAT1 staining in the islets, the antibodies to these proteins selectively stained astrocytes and GABAergic nerve endings, respectively, in the brain.…”

Section: Control Brain Sectionssupporting

confidence: 57%

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

Gammelsaeter

Frøyland

Aragón

et al. 2004

Journal of Cell Science

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To elucidate the possible roles of the CNS neurotransmitters glycine and GABA in neuroendocrine paracrine signalling, we investigated their localizations, and those of their transport proteins, by confocal immunofluorescence and quantitative post-embedding immuno-electron microscopy in the pancreatic islets of Langerhans. We show that A-cells contain glycine in synaptic-like microvesicles as well as in secretory granules. A-cells express the macromolecules necessary to: (1) concentrate glycine within both organelle types before release (the vesicular GABA/glycine transporter VGAT=VIAAT); and to (2) take up the transmitter from the extracellular space (the plasma membrane glycine transporter GLYT2). Also B-cells have glycine in their microvesicles and granules, but the microvesicle/cytosol ratio is lower than in A-cells, consistent with the presence of GABA (which competes with glycine for vesicular uptake) in the cytosol at a much higher concentration in B-cells than in A-cells. Both A- and B-cells contain GABA in their microvesicles and secretory granules, and the membranes of the two organelle types contain VGAT in both cell types. A-cells as well as B-cells express a plasma membrane transporter GAT3 that mediates uptake of GABA. The localization of VGAT in the cores of A-cell secretory granules, and in the secretory granule membranes in both cell types, indicates novel aspects of the mechanisms for release of glycine and GABA. The discovery that both A- and B-cells possess the molecular machinery for the evoked release of both glycine and GABA from synaptic-like microvesicles suggests that both of the principal inhibitory transmitters in the brain participate in paracrine signalling in the pancreas.

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Section: Control Brain Sectionssupporting

confidence: 57%

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

Gammelsaeter

Frøyland

Aragón

et al. 2004

Journal of Cell Science

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“…Among the high-affinity GABA transporters, GAT-2 is the only one expressed in peripheral tissues in addition to brain and retina (Borden et al, 1992;Liu et al, 1993;Ikegaki et al, 1994), therefore it is very likely that GAT-2 is also the ALA transporter in murine mammary adenocarcinoma cells. However, both the regulation of gene expression and substrate specificity in neoplastic cells may be different from the corresponding normal tissue; if so, these differences may be exploited to enhance ALA-PDT selectivity.…”

Section: Experimental Therapeuticsmentioning

confidence: 99%

δ-Aminolevulinic acid transport in murine mammary adenocarcinoma cells is mediated by beta transporters

et al. 2002

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d-aminolevulinic acid, the precursor of porphyrin biosynthesis has been used to induce the endogenous synthesis of the photosensitiser protoporphyrin IX for photodynamic therapy in the treatment of various tumours. The aim of this work was to characterise the d-aminolevulinic acid transport system in the murine mammary adenocarcinoma cell line LM3 using 14 C-daminolevulinic acid, to finally improve d-aminolevulinic acid incorporation in mammalian cells. Our results showed that daminolevulinic acid is incorporated into these cells by two different mechanisms, passive diffusion which is important at the beginning of the incubation, and active transport. Specificity assays suggested that the transporter involved in d-aminolevulinic acid incorporation is a BETA transporter, probably GAT-2.

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“…Similar requirements of repetitive stimulation for the induction of GABA B -mediated responses have been reported in other neurons (Johnson et al 1992;Saitoh et al 2004). The abundantly expressed GABA transporters in the GP might regulate the extent of GABA B receptor activation (Chen and Yung 2003;Ikegaki et al 1994;Yasumi et al 1997).…”

Section: Responses To Intra-pallidal Repetitive Stimulationmentioning

confidence: 99%

Synaptically Released GABA Activates Both Pre- and Postsynaptic GABA_B Receptors in the Rat Globus Pallidus

Kaneda¹,

Kita²

2005

Journal of Neurophysiology

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Kaneda, Katsuyuki and Hitoshi Kita. Synaptically released GABA activates both pre-and postsynaptic GABA B receptors in the rat globus pallidus. J Neurophysiol 94: 1104 -1114, 2005; doi:10.1152/jn.00255.2005. The globus pallidus (GP) contains abundant GABAergic synapses and GABA B receptors. To investigate whether synaptically released GABA can activate pre-and postsynaptic GABA B receptors in the GP, physiological recordings were performed using rat brain slice preparations. Cell-attached recordings from GABA A antagonist-treated preparations revealed that repetitive local stimulation induced a GABA B antagonist-sensitive pause in spontaneous firings of GP neurons. Whole cell recordings revealed that the repetitive stimulation evoked fast excitatory postsynaptic potentials followed by a slow inhibitory postsynaptic potential (IPSP) in GP neurons. The slow IPSP was insensitive to a GABA A receptor antagonist, increased in amplitude with the application of ionotropic glutamate receptor antagonists, and was suppressed by the GABA B antagonist CGP55845. The reversal potential of the slow IPSP was close to the potassium equilibrium potential. These results suggest that synaptically released GABA activated postsynaptic GABA B receptors and induced the pause and the slow IPSP. On the other hand, in the neurons that were treated to block postsynaptic GABA B responses, CGP55845 increased the amplitudes of repetitive local stimulationinduced GABA A -mediated inhibitory postsynaptic currents (IPSCs) but not the ionotropic glutamate-mediated excitatory postsynaptic currents. Moreover, the GABA B receptor specific agonist baclofen reduced the frequency of miniature IPSCs without altering their amplitude distributions. These results suggest that synaptically released GABA also activated presynaptic GABA B autoreceptors, resulting in decreased GABA release in the GP. Together, we infer that both pre-and postsynaptic GABA B receptors may play crucial roles in the control of GP neuronal activity.

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Production of specific antibodies against GABA transporter subtypes (GAT1, GAT2, GAT3) and their application to immunocytochemistry

Cited by 123 publications

References 15 publications

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

δ-Aminolevulinic acid transport in murine mammary adenocarcinoma cells is mediated by beta transporters

Synaptically Released GABA Activates Both Pre- and Postsynaptic GABA_B Receptors in the Rat Globus Pallidus

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Production of specific antibodies against GABA transporter subtypes (GAT1, GAT2, GAT3) and their application to immunocytochemistry

Cited by 123 publications

References 15 publications

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay

δ-Aminolevulinic acid transport in murine mammary adenocarcinoma cells is mediated by beta transporters

Synaptically Released GABA Activates Both Pre- and Postsynaptic GABAB Receptors in the Rat Globus Pallidus

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Synaptically Released GABA Activates Both Pre- and Postsynaptic GABA_B Receptors in the Rat Globus Pallidus