Abstract
We studied 3H-glycine and 3H-strychnine specific binding to glycine receptor (GlyR) in intact isolated frog retinas. To avoid glycine binding to glycine uptake sites, experiments were performed at low ligand concentrations in a sodium-free medium. The binding of both radiolabeled ligands was saturated. Scatchard analysis of bound glycine and strychnine revealed a KD of 2.5 and 2.0 μM, respectively. Specific binding of glycine was displaced by β-alanine, sarcosine, and strychnine. Strychnine binding was displaced 50% by glycine, and sarcosine. Properties of the strychnine-binding site in the GlyR were modified by sarcosine. Binding of both radioligands was considerably reduced by compounds that inhibit or activate adenylate cyclase and increased cAMP levels. A phorbol ester activator of PKC remarkably decreased glycine and strychnine binding. These results suggest modulation of GlyR in response to endogenous activation of protein kinases A and C, as well as protein phosphorylation modulating GlyR function in retina.
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B. Matzenbach Y. Maulet L. Sefton B. Courtier P. Avner J. L. Guénet H. Betz (1994) ArticleTitleStructural analysis of mouse glycine receptor α subunits genes: Identification and chromosomal localization of a novel variant, α4 J. Biol. Chem. 269 2607–2612 Occurrence Handle1:CAS:528:DyaK2cXivFWjs7k%3D Occurrence Handle7507926
B. Laube G. Maksay R. Schemm H. Betz (2002) ArticleTitleModulation of glycine receptor function: A novel approach for therapeutic intervention at inhibitory synapses? Trends Pharmacol. Sci. 23 519–527 Occurrence Handle1:CAS:528:DC%2BD38XotF2js70%3D Occurrence Handle12413807
A. Ruiz-Gomez M. L. Vaello F. Valdivieso F. Mayor SuffixJr. (1991) ArticleTitlePhosphorylation of the 48-kDa subunit of the glycine receptor by protein kinase C J. Biol. Chem. 266 559–566 Occurrence Handle1:CAS:528:DyaK3MXhtFOms70%3D Occurrence Handle1845981
Y. M. Song L. Y. Huang (1990) ArticleTitleModulation of glycine receptor chloride channels by cAMP-dependent protein kinase in spinal trigeminal neurons Nature 249 1580–1585
H. Inomata J. Nabekura N. Akaike (1993) ArticleTitleSuppression of taurine response in acutely dissociated substantia nigra neurons by intracellular cyclic AMP Brain Res. 615 347–350 Occurrence Handle1:CAS:528:DyaK3sXltlaisbY%3D Occurrence Handle8395961
N. Agopyan N. Tokutomi N. Akaike (1993) ArticleTitleProtein kinase A-mediated phosphorylation reduces only the fast sensitizing glycine current in acutely dissociated ventromedial hypothalamic neurons Neuroscience 56 605–615 Occurrence Handle1:CAS:528:DyaK2cXks1erug%3D%3D Occurrence Handle7504795
M. L. Vaello A. Ruiz-Gómez J. Lerma F. Mayor SuffixJr. (1994) ArticleTitleModulation of inhibitory glycine receptors by phosphorylation by protein kinase C and cAMP-dependent protein kinase J. Biol. Chem. 269 2002–2008 Occurrence Handle1:CAS:528:DyaK2cXps1yrtQ%3D%3D Occurrence Handle8294452
B. Schönrock J. Bormann (1995) ArticleTitleModulation of hippocampal glycine receptor channels by protein kinase C Neuroreport 6 301–304 Occurrence Handle7756615
R. E. Marc (1989) ArticleTitleThe role of glycine in mammalian retina Prog. Retinal Res. 8 119–158
U. Greferath J. H. Brandstatter H. Wässle J. Kirsch J. Kuhse U. Grünert (1994) ArticleTitleDifferential expression of glycine receptor subunits in the retina of the rat: a study using immunohistochemistry and in situ hybridization Visual Neurosci. 11 721–729
U. Grünert H. Wässle (1993) ArticleTitleImmunohistochemical localization of glycine receptors in the mammalian retina J. Comp. Neurol. 335 523–537 Occurrence Handle8227534
S. Haverkamp U. Müller K. Harvey R. Harvey H. Betz H. Wässle (2003) ArticleTitleDiversity of glycine receptors in the mouse retina: localization of the α3 subunit J. Comp. Neurol. 465 524–539 Occurrence Handle1:CAS:528:DC%2BD3sXoslWqsbs%3D Occurrence Handle12975813
J. F. Smiley S. Yazulla (1990) ArticleTitleGlycinergic contacts in the outer plexiform layer of the Xenopus laevis retina characterized by antibodies to glycine, GABA and glycine receptors J. Comp. Neurol. 299 375–388 Occurrence Handle1:CAS:528:DyaK3MXit1Kjsrk%3D Occurrence Handle2172330
J. F. Pérez-León S. Salceda (2000) ArticleTitleImmunohistochemical localization of the glycine receptor in synaptic layers and glial cells of the retina Sensory Neuron 3 71–81
M. J. Neal (1976) ArticleTitleAmino acid transmitter substances in the vertebrate retina J. Gen. Pharmacol. 7 321–332 Occurrence Handle1:CAS:528:DyaE2sXhsVajsA%3D%3D
R. Salceda (1989) ArticleTitleUptake and K+-stimulated release of 14C-glycine from frog retinal synaptosomal fractions Neurochem. Res. 14 49–54 Occurrence Handle1:CAS:528:DyaL1MXhsVymu7g%3D Occurrence Handle2710278
G. A. Johnson L. L. Iversen (1971) ArticleTitleGlycine uptake in the control nervous system slices and homogenates: Evidence for different uptake mechanisms in spinal cord and cerebral cortex J. Neurochem. 18 1951–1961 Occurrence Handle4399087
H. Kishimoto J. R. Simon M. H. Aprison (1981) ArticleTitleDetermination of the equilibrium dissociation constants and number of glycine binding sites in several areas of the rat central nervous system, using a sodium-independent system J. Neurochem. 37 1015–1024 Occurrence Handle1:CAS:528:DyaL3MXlvVegtLg%3D Occurrence Handle6275030
A. B. Young S. H. Snyder (1974) ArticleTitleStrychnine binding in rat spinal cord membranes associated with the synaptic glycine receptor: cooperativity of glycine interactions Mol. Pharmacol. 10 790–809 Occurrence Handle1:CAS:528:DyaE2MXjvFWhtQ%3D%3D
J. A. Pérez-León R. Salceda (1995) ArticleTitleDifferent specific binding sites of 3H-glycine and 3H-strychnine in synaptosomal membranes isolated from frog retina Neurochem. Res. 20 915–922 Occurrence Handle8587649
V. Schmieden H. Betz (1995) ArticleTitlePharmacology of the inhibitory glycine receptor: agonist and antagonist actions of amino acids and piperidine carboxylic acid compounds Mol. Pharmacol. 48 919–927 Occurrence Handle1:CAS:528:DyaK2MXps1amtrg%3D Occurrence Handle7476923
L. A. Raymond C. D. Blackstone R. L. Hunganir (1993) ArticleTitlePhosphorylation of amino acid neurotransmitter receptors in synaptic plasticity Trends. Neurosci. 16 147–153 Occurrence Handle1:CAS:528:DyaK3sXks1eisbY%3D Occurrence Handle7682348
Y. Gu L. Y. M. Huang (1998) ArticleTitleCross-modulation of glycine activated Cl− channels by protein kinase C and cAMP-dependent protein kinases in the rat J. Physiol. 506 331–339 Occurrence Handle1:CAS:528:DyaK1cXhtFWqsbc%3D Occurrence Handle9490862
F. A. Albarran J. P. Roa R. Navarrete R. Castillo F. Nualart L. G. Aguayo (2001) ArticleTitleEffect of protein kinase C activation on the glycine evoked Cl− current in spinal cord neurons Brain Res. 902 1–10 Occurrence Handle1:CAS:528:DC%2BD3MXjslejurs%3D Occurrence Handle11376589
Y. Han M. M. Slaughter (1998) ArticleTitleProtein kinases modulate two glycine currents in salamander retinal ganglion cells J. Physiol. 508 681–690 Occurrence Handle1:CAS:528:DyaK1cXjtlShtbY%3D Occurrence Handle9518725
C. Galli O. Mencci A. Scorziello T. M. Werge P. Calissano G. Schettini (1995) ArticleTitleApoptosis of cerebellar granule cells is blocked by high KCl, forskolin, and IGF-1 through distinct mechanisms of action: the involvement of intracellular calcium and RNA synthesis J. Neurosci. 15 1172–1179 Occurrence Handle1:CAS:528:DyaK2MXjvVCgtrg%3D Occurrence Handle7532699
S. H. Shafer S. H. Phelps C. L. Williams (1998) ArticleTitleReduced DNA synthesis and cell viability in small lung carcinoma by treatment with cyclic AMP phosphodiesterase inhibitors Biochem. Pharmacol. 56 1229–1236 Occurrence Handle1:CAS:528:DyaK1cXnt1OhtL8%3D Occurrence Handle9802335
D. R. Sibley J. L. Benovic M. G. Caron R. J. Lefkowitz (1987) ArticleTitleRegulation of transmembrane signaling by receptor phosphorylation Cell 48 913–922 Occurrence Handle1:CAS:528:DyaL1cXjt1Shtg%3D%3D Occurrence Handle3030559
T. S. Rao J. A. Cler M. R. Emmett S. J. Mick S. Iyengar P. L. Wood (1990) ArticleTitleGlycine, glycinamide and D-serine act as a positive modulator of signal transduction at the NMDA receptor in vivo: Differential effects on mouse cerebellar cyclic guanosine monophosphate levels Neuropharmacol. 29 1075–1080 Occurrence Handle1:CAS:528:DyaK3MXjs1Crug%3D%3D
M. Montiel J. Quesada E. Jiménez (2004) ArticleTitleActivation of second messenger-dependent protein kinases induces muscarinic acetylcholine receptor desensitization in rat thryroid epithelial cells Mol. Cell Endocrinol. 223 35–41 Occurrence Handle1:CAS:528:DC%2BD2cXlvFyrtLs%3D Occurrence Handle15279909
Y. Namkung D. R. Sibley (2004) ArticleTitleProtein kinase C mediates phosphorylation, desensitization and trafficking of the D2 dopamine receptor J. Biol. Chem. 279 49533–49541 Occurrence Handle1:CAS:528:DC%2BD2cXpslKitbY%3D Occurrence Handle15347675
J. Nabekura T. Omura N. Akaike (1996) ArticleTitleAlpha 2 adrenoceptor potentiates glycine receptor-mediated taurine response through protein kinase A in rat substantia nigra neurons Neurophysiol. 76 2447–2454 Occurrence Handle1:CAS:528:DyaK28XmvFKgtro%3D
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Salceda, R., Aguirre-Ramirez, M. Characterization of Strychnine-sensitive Glycine Receptor in the Intact Frog Retina: Modulation by Protein Kinases. Neurochem Res 30, 411–416 (2005). https://doi.org/10.1007/s11064-005-2616-1
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DOI: https://doi.org/10.1007/s11064-005-2616-1