Effects of intrahippocampal administration of the phosphatase inhibitor okadaic acid: Dual effects on memory formation

Vianna, Monica R.M.; Coitinho, Adriana; Izquierdo, Luciana; Izquierdo, Ivan

doi:10.1590/S1980-57642010DN40100004

Abstract

Protein phosphorylation mediated by serine-threonine kinases in the hippocampus is crucial to the synaptic modifications believed to underlie memory formation. The role of phosphatases has been the focus of comparatively little study. Objectives: Here we evaluate the contribution of the serine-threonine protein phosphatases 1 and 2A (PP1, PP2A) on memory consolidation. Methods: We used immediate post-training bilateral hippocampal infusions of okadaic acid (OA, 0.01 and 10 pmol/side), a potent inhibitor of PP1 and PP2A, and measured short- [3 h] and long-term memory [24 h] (STM, LTM) of step-down inhibitory avoidance. Results: At the lower dose, OA inhibited both STM and LTM whereas at the higher dose it instead enhanced LTM. Pre-test infusion of these two doses of OA had no effect on retrieval. Conclusions: These two doses of OA are known to selectively inhibit PP1 and PP2A respectively. These findings point to the importance of these enzymes in memory formation and also suggest a deleterious influence of endogenous hippocampal PP2A on LTM formation.

Key words:
hippocampus; PP1; PP2A; okadaic acid; short-term memory; long-term memory

Resumo

A fosforilação de proteínas mediada por serina-treonina quinases no hipocampo é crucial para as modificações sinápticas que se acredita sejam necessárias para a formação de memórias. O papel das fosfatases tem sido comparativamente pouco estudado. Objetivos: Aqui avaliamos a contribuição das fosfatases serina-treonina 1 e 2 (PP1, PP2A) sobre a consolidação da memória. Métodos: Usamos infusões imediatamente após o treino de ácido okadaico (OA, 0.01 e 10 pmol/lado), um potente inibidor de PP1 e medimos memória de curta [3 h] e longa duração [24 h] (STM, LTM) de esquiva inibitória de evitar descer de uma plataforma. Resultados: Na dose menor, OA inibiu tanto STM como LTM. Na dose maior, produziu, em vez disso, uma melhora da LTM. A infusão pré-teste de qualquer uma das duas doses de OA não teve efeito sobre a evocação. Conclusões: Estas duas doses de OA são conhecidas por inibir seletivamente PP1 a PP2 respectivamente. Estes resultados apontam à importância das duas enzimas na formação de memória e sugerem, adicionalmente, uma influência deletérea da PP2A endógena sobre a formação de LTM.

Palavras-chave:
hipocampo; PP1; PP2A; ácido ocadaico; memória de curto prazo; memória de longo prazo

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References

¹
Winder DG, Sweatt JD Roles of serine/threonine phosphatases in hippocampal synaptic plasticity. Nature Rev Neurosci 2001; 2:461-474.
²
Izquierdo I, Bevilaqua LRM, Rossato JI, Bonini JS, Medina JH, Cammarota M. Different molecular cascades in different sites of the brain control consolidation. Trends Neurosci 2006;29: 496-505.
³
Cammarota M, Bernabeu R, Levi de Stein M, Izquierdo I, Medina JH. Learning-specific, time-dependent increases in hippocampal Ca2+/calmodulin-dependent protein kinase II activity and AMPA GluR1 subunit immunoreactivity. Eur J Neurosci 1998;10:2669-2676.
⁴
Bevilaqua LR, Medina JH, Cammarota M, Izquierdo I. Memory consolidation induces N-methyl-d-aspartic acid receptor and Ca(2+)/calmodulin-dependent protein kinase II-dependent modifications in alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor properties. Neurosci 2005;136: 397-403.
⁵
Bernabeu R, Bevilaqua L, Ardenghi P, Bromberg E, Schmitz P, Bianchin M, Izquierdo I, Medina JH. Involvement of hippocampal D1/D5 receptor - cAMP signaling pathways in a late memory consolidation phase of an aversively-motivated task in rats. Proc Natl Acad Sci USA 1997;94:7041-7046.
⁶
Samuels IS, Karlo JC, Faruzzi AN, et al. Deletion of ERK2 mitogen-activated protein kinase identifies its key roles in cortical neurogenesis and cognitive function. J Neurosci 2008; 28:6983-6995.
⁷
Schrader LA, Ren Y, Cheng F, Bui D, Sweatt JD, Anderson AE. Kv4.2 is a locus for PKC and ERK/MAPK cross-talk. Biochem J 2009;417:705-715.
⁸
Paratcha G, Furman M, Bevilaqua L, et al. Involvement of hippocampal PKCbI isoform in the early phase of memory formation of an inhibitory avoidance learning. Brain Res 2000;855:199-205.
⁹
Holahan M, Routtenberg A. The protein kinase C phosphorylation site on GAP-43 differentially regulates information storage. Hippocampus 2008;18:1099-1102.
¹⁰
Riedel G, Platt B. From messengers to molecules. New York: Kluwer Academic; 2004: 614.
¹¹
Zhao W, Bennett P, Sedman GL, Ng KT. The impairment of long-term memory formation by the phosphatase inhibitor okadaic acid. Brain Res Bull 1995;36:557-561.
¹²
Bennett PC, Moutsoulas P, Lawen A, Perini E, Ng KT. Novel effects on memory observed following unilateral intracranial administration of okadaic acid, cyclosporine A, FK506 and [VeVal4]CyA. Brain Res 2003;988:56-68.
¹³
Mansuy IM, Shenolikar S. Protein serine/threonine phosphatases in neuronal plasticity and disorders of learning and memory. Trends Neurosci 2006;29:679-686.
¹⁴
Stefan MI, Edelstein SJ, Le Novère N. An allosteric model of calmodulin explains differential activation of PP2B and CaMKII. Proc Natl Acad Sci USA 2008;105:10768-10773.
¹⁵
Wang LY, Orser BA, Brautigan DL, MacDonald JF. Regulation of NMDA receptors in cultured hippocampal neurons by protein phosphatases 1 and 2A. Nature 1994;369:230-232.
¹⁶
Woo NH, Nguyen PV. Silent metaplasticity of the late phase of long-term potentiation requires protein phosphatases. Learn Mem 2002;9:202-213.
¹⁷
Gruart A, Muñoz MD, Delgado-García JM. Involvement of the CA3-CA1 synapse in the acquisition of associative learning in behaving mice. J Neurosci 2006;26:1077-1087.
¹⁸
Whitlock JR, Heynen AJ, Shuler MG, Bear MF. Learning induces long-term potentiation in the hippocampus. Science 2006;313:1093-1097.
¹⁹
Bennett PC, Zhao W, Lawan A, Ng KT. Cyclosporin A, an inhibitor of calcineurin, impairs memory formation in day-old chicks. Brain Res 1996;730:107-117.
²⁰
Sun L, Liu SY, Zhow XW, Wang XC, Liu R, Wang Q, Wang JZ. Inhibition of protein phosphatase 2A- and protein phosphatase 1-induced tau hyperphosphorylation and impairment of spatial memory retention in rats. Neurosci 2003;118:1175-1182.
²¹
Bennett PC, Zhao W, Ng KT. Concentration-dependent effects of protein phosphatases (PP) inhibitors implicate PP1 and PP2A in different stages of memory formation. Neurobiol Learn Mem 2001;75:91-110. 22.
²²
Winder DG, Mansuy IM, Osman M, Moallem TM, Kandel ER. Genetic and pharmacological evidence for a novel, intermediate phase of long-term potentiation suppressed by calcineurin. Cell 1998;92:25-37.
²³
Mansuy IM, Mayford M, Jacob B, Kandel ER, Bach ME. Restricted and regulated overexpression reveals calcineurin as a key component in the transition from short-term to long-term memory. Cell 1998;92:39.
²⁴
Malleret G, Haditsch U, Genoux D, et al. Inducible and reversible enhancement of learning, memory, and long-term potentiation by genetic inhibition of Calcineurin. Cell 2001; 104:675-686.
²⁵
Genoux D, Haditsch U, Knobloch M, Michalon A, Storm D, Mansuy IM. Protein phosphatase 1 is a molecular constraint on learning and memory. Nature 2002;418:929-930.
²⁶
Jouvenceau A, Hédou G, Potier B, Kollen M, Dutar P, Mansuy IM. Partial inhibition of PP1 alters bidirectional synaptic plasticity in the hippocampus. Eur J Neurosci 2006;24:564-572.
²⁷
Izquierdo I, Barros DM, Mello e Souza T, Souza MM, Izquierdo LA, Medina JH. Mechanisms for memory types differ. Nature 1998;393:635-636.
²⁸
McGaugh JL. Memory: a century of consolidation. Science 2000;287:248-251.
²⁹
Cohen P, Alemany S, Hemmings BA, Resink TJ, Strålfors P, Tung HY. Protein phosphatase-1 and protein phosphatase-2A from rabbit skeletal muscle. Meth Enzymol 1988;159:390-408.
³⁰
Grahame-Hardie D. Protein phosphorylation: a practical approach, Oxford Academic Press: Oxford; 1999.
³¹
Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates. Academic Press: San Diego; 1986.
³²
Vianna MR, Alonso M, Viola H, et al. Role of hippocampal signaling pathways in long-term memory formation of a nonassociative learning task in the rat. Learn Mem 2000;7:333-340.
³³
Arendt T, Holzer M, Fruth R, Bruckner MK, Gartner U. Paired helical filament-like phosphorylation of tau, deposition of beta/A4-amyloid and memory impairment in rat induced by chronic inhibition of phosphatase 1 and 2A. Neuroscience 1995;69:691-698.
³⁴
Silva AJ, Josselyn SA. The molecules of forgetfulness. Nature 2002;418:929-930.
³⁵
Izquierdo I. A arte de esquecer. Rio de Janeiro, Vieira & Lent; 2004.
³⁶
Lopez-Salon M, Alonso M, Vianna, MR, Viola H, Mello e Souza T, Izquierdo I, Pasquini JM, Medina JH. The ubiquitin-proteasome cascade is required for mammalian long-term memory formation. Eur J Neurosci 2001;14:1820-1826.

Publication Dates

Publication in this collection
Jan-Mar 2010

History

Received
16 Sept 2009
Accepted
20 Feb 2010

This is an open-access article distributed under the terms of the Creative Commons Attribution License

[1] ¹
Winder DG, Sweatt JD Roles of serine/threonine phosphatases in hippocampal synaptic plasticity. Nature Rev Neurosci 2001; 2:461-474.

[2] ²
Izquierdo I, Bevilaqua LRM, Rossato JI, Bonini JS, Medina JH, Cammarota M. Different molecular cascades in different sites of the brain control consolidation. Trends Neurosci 2006;29: 496-505.

[3] ³
Cammarota M, Bernabeu R, Levi de Stein M, Izquierdo I, Medina JH. Learning-specific, time-dependent increases in hippocampal Ca2+/calmodulin-dependent protein kinase II activity and AMPA GluR1 subunit immunoreactivity. Eur J Neurosci 1998;10:2669-2676.

[4] ⁴
Bevilaqua LR, Medina JH, Cammarota M, Izquierdo I. Memory consolidation induces N-methyl-d-aspartic acid receptor and Ca(2+)/calmodulin-dependent protein kinase II-dependent modifications in alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor properties. Neurosci 2005;136: 397-403.

[5] ⁵
Bernabeu R, Bevilaqua L, Ardenghi P, Bromberg E, Schmitz P, Bianchin M, Izquierdo I, Medina JH. Involvement of hippocampal D1/D5 receptor - cAMP signaling pathways in a late memory consolidation phase of an aversively-motivated task in rats. Proc Natl Acad Sci USA 1997;94:7041-7046.

[6] ⁶
Samuels IS, Karlo JC, Faruzzi AN, et al. Deletion of ERK2 mitogen-activated protein kinase identifies its key roles in cortical neurogenesis and cognitive function. J Neurosci 2008; 28:6983-6995.

[7] ⁷
Schrader LA, Ren Y, Cheng F, Bui D, Sweatt JD, Anderson AE. Kv4.2 is a locus for PKC and ERK/MAPK cross-talk. Biochem J 2009;417:705-715.

[8] ⁸
Paratcha G, Furman M, Bevilaqua L, et al. Involvement of hippocampal PKCbI isoform in the early phase of memory formation of an inhibitory avoidance learning. Brain Res 2000;855:199-205.

[9] ⁹
Holahan M, Routtenberg A. The protein kinase C phosphorylation site on GAP-43 differentially regulates information storage. Hippocampus 2008;18:1099-1102.

[10] ¹⁰
Riedel G, Platt B. From messengers to molecules. New York: Kluwer Academic; 2004: 614.

[11] ¹¹
Zhao W, Bennett P, Sedman GL, Ng KT. The impairment of long-term memory formation by the phosphatase inhibitor okadaic acid. Brain Res Bull 1995;36:557-561.

[12] ¹²
Bennett PC, Moutsoulas P, Lawen A, Perini E, Ng KT. Novel effects on memory observed following unilateral intracranial administration of okadaic acid, cyclosporine A, FK506 and [VeVal4]CyA. Brain Res 2003;988:56-68.

[13] ¹³
Mansuy IM, Shenolikar S. Protein serine/threonine phosphatases in neuronal plasticity and disorders of learning and memory. Trends Neurosci 2006;29:679-686.

[14] ¹⁴
Stefan MI, Edelstein SJ, Le Novère N. An allosteric model of calmodulin explains differential activation of PP2B and CaMKII. Proc Natl Acad Sci USA 2008;105:10768-10773.

[15] ¹⁵
Wang LY, Orser BA, Brautigan DL, MacDonald JF. Regulation of NMDA receptors in cultured hippocampal neurons by protein phosphatases 1 and 2A. Nature 1994;369:230-232.

[16] ¹⁶
Woo NH, Nguyen PV. Silent metaplasticity of the late phase of long-term potentiation requires protein phosphatases. Learn Mem 2002;9:202-213.

[17] ¹⁷
Gruart A, Muñoz MD, Delgado-García JM. Involvement of the CA3-CA1 synapse in the acquisition of associative learning in behaving mice. J Neurosci 2006;26:1077-1087.

[18] ¹⁸
Whitlock JR, Heynen AJ, Shuler MG, Bear MF. Learning induces long-term potentiation in the hippocampus. Science 2006;313:1093-1097.

[19] ¹⁹
Bennett PC, Zhao W, Lawan A, Ng KT. Cyclosporin A, an inhibitor of calcineurin, impairs memory formation in day-old chicks. Brain Res 1996;730:107-117.

[20] ²⁰
Sun L, Liu SY, Zhow XW, Wang XC, Liu R, Wang Q, Wang JZ. Inhibition of protein phosphatase 2A- and protein phosphatase 1-induced tau hyperphosphorylation and impairment of spatial memory retention in rats. Neurosci 2003;118:1175-1182.

[21] ²¹
Bennett PC, Zhao W, Ng KT. Concentration-dependent effects of protein phosphatases (PP) inhibitors implicate PP1 and PP2A in different stages of memory formation. Neurobiol Learn Mem 2001;75:91-110. 22.

[22] ²²
Winder DG, Mansuy IM, Osman M, Moallem TM, Kandel ER. Genetic and pharmacological evidence for a novel, intermediate phase of long-term potentiation suppressed by calcineurin. Cell 1998;92:25-37.

[23] ²³
Mansuy IM, Mayford M, Jacob B, Kandel ER, Bach ME. Restricted and regulated overexpression reveals calcineurin as a key component in the transition from short-term to long-term memory. Cell 1998;92:39.

[24] ²⁴
Malleret G, Haditsch U, Genoux D, et al. Inducible and reversible enhancement of learning, memory, and long-term potentiation by genetic inhibition of Calcineurin. Cell 2001; 104:675-686.

[25] ²⁵
Genoux D, Haditsch U, Knobloch M, Michalon A, Storm D, Mansuy IM. Protein phosphatase 1 is a molecular constraint on learning and memory. Nature 2002;418:929-930.

[26] ²⁶
Jouvenceau A, Hédou G, Potier B, Kollen M, Dutar P, Mansuy IM. Partial inhibition of PP1 alters bidirectional synaptic plasticity in the hippocampus. Eur J Neurosci 2006;24:564-572.

[27] ²⁷
Izquierdo I, Barros DM, Mello e Souza T, Souza MM, Izquierdo LA, Medina JH. Mechanisms for memory types differ. Nature 1998;393:635-636.

[28] ²⁸
McGaugh JL. Memory: a century of consolidation. Science 2000;287:248-251.

[29] ²⁹
Cohen P, Alemany S, Hemmings BA, Resink TJ, Strålfors P, Tung HY. Protein phosphatase-1 and protein phosphatase-2A from rabbit skeletal muscle. Meth Enzymol 1988;159:390-408.

[30] ³⁰
Grahame-Hardie D. Protein phosphorylation: a practical approach, Oxford Academic Press: Oxford; 1999.

[31] ³¹
Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates. Academic Press: San Diego; 1986.

[32] ³²
Vianna MR, Alonso M, Viola H, et al. Role of hippocampal signaling pathways in long-term memory formation of a nonassociative learning task in the rat. Learn Mem 2000;7:333-340.

[33] ³³
Arendt T, Holzer M, Fruth R, Bruckner MK, Gartner U. Paired helical filament-like phosphorylation of tau, deposition of beta/A4-amyloid and memory impairment in rat induced by chronic inhibition of phosphatase 1 and 2A. Neuroscience 1995;69:691-698.

[34] ³⁴
Silva AJ, Josselyn SA. The molecules of forgetfulness. Nature 2002;418:929-930.

[35] ³⁵
Izquierdo I. A arte de esquecer. Rio de Janeiro, Vieira & Lent; 2004.

[36] ³⁶
Lopez-Salon M, Alonso M, Vianna, MR, Viola H, Mello e Souza T, Izquierdo I, Pasquini JM, Medina JH. The ubiquitin-proteasome cascade is required for mammalian long-term memory formation. Eur J Neurosci 2001;14:1820-1826.

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