Intra-CA1 administration of FK-506 (tacrolimus) in rat impairs learning and memory in an inhibitory avoidance paradigm

Document Type : Original Article


1 Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran

2 Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran. Department of Pharmacology, Kashan University of Medical Sciences, Kashan, Iran


Objective(s): Calcineurin (CN) is a main phosphatase and a critical regulator of cellular pathways for learning, memory, and plasticity. The FK-506 (tacrolimus),a phosphatase inhibitor, is a fungal-derived agent and a common immune suppressant extensively used for tissue transplantation. To further clarify the role of CN in different stages oflearning and memory the main aim of this study was to evaluate the role of FK-506 in an inhibitory avoidance model.
Materials and Methods:Using different doses of FK-506 (0.5, 5, and 50 nM) in the CA1 of hippocampus at different times (before, after the training and also before the test), the effect of drug was evaluated in a step-through inhibitory avoidance paradigm.The latency of entering to the dark compartment was considered as a criterion for memory.
Results:  Thepre-training intra-CA1 injections of FK-506 impaired inhibitory avoidance (IA) learning acquisition. In addition, thepost-training intra-CA1 injections of FK-506 at 1, 2, and 3 hr relative to training impaired memory consolidation. Moreover, thepre-test intra-CA1 injections of FK-506 impaired memory retrieval.
Conclusion:These findings suggest that the FK-506 selectively interferes with acquisition, retention, and retrieval of information processing in CA1 of hippocampus. Given the crucial role of CN in common signaling pathway of higher functions such as memory performance and cognition, in future it would be a probable therapeutic target in the treatment of a wide verity of neurological conditions involving memory.


1. Bliss TV, Collingridge GL. A synaptic model of memory: long-term potentiation in the hippocampus. Nature 1993; 361:31-39.
2 .Morris RG, Moser EI, Riedel G, Martin SJ, Sandin J, Day M, et al. Elements of a neurobiological theory of the hippocampus: the role of activity-dependent synaptic plasticity in memory. Philosophical transactions of the Royal Society of London Series. Biol Sci 2003; 358:773-786.
3. Lynch MA. Long-term potentiation and memory. Physiol Rev 2004; 84:136-87.
4. Abel T, Martin KC, Bartsch D, Kandel ER. Memory suppressor genes: inhibitory constraints on the storage of long-term memory. Science 1998; 279:338-341.
5. Taglialatela G, Hogan D, Zhang WR, Dineley KT. Intermediate- and long-term recognition memory deficits in Tg2576 mice are reversed with acute calcineurin inhibition. Behav Brain Res 2009; 200:95-99.
6 .Rozkalne A, Hyman BT, Spires-Jones TL. Calcineurin inhibition with FK506 ameliorates dendritic spine density deficits in plaque-bearing Alzheimer model mice. Neurobiol Dis  2011; 41:650-654.
7. Snyder SH, Sabatini DM. Immunophilins and the nervous system. Nat Med  1995; 1:32-37.
8. Lee SH, Kim BC, Yang DH, Park MS, Choi SM,               Kim MK, et al. Calcineurin inhibitor-mediated bilateral hippocampal injury after bone marrow transplantation. J Neurol 2008; 255:929-931.
9 .Wijdicks EF, Wiesner RH, Dahlke LJ, Krom RA. FK506-induced neurotoxicity in liver transplantation. Ann Neurol 1994; 69:105-111.
10. Bennett PC, Moutsoulas P, Lawen A, Perini E, Ng KT. Novel effects on memory observed following unilateral intracranial administration of okadaic acid, cyclosporin A, FK506 and [MeVal4]CyA. Brain Res 2003; 988:56-6,501-598.
11. Ho S, Clipstone N, Timmermann L, Northrop J, Graef I, Fiorentino D, et al. The mechanism of              action of cyclosporin A and FK506. Clin Immunol Immunopathol 1996; 80:S40-45.
12. Snyder SH, Lai MM, Burnett PE. Immunophilins in the nervous system. Neuron 1998; 21:283-294.
13. Tanaka K, Hori K, Wada-Tanaka N, Nomura M, Ogawa N. FK506 ameliorates the discrimination learning impairment due to preventing the rarefaction of white matter induced by chronic cerebral hypoperfusion in rats. Brain Res 2001; 906:184-189.
14. Spires-Jones TL, Kay K, Matsouka R, Rozkalne A, Betensky RA, Hyman BT. Calcineurin inhibition with systemic FK506 treatment increases dendritic branching and dendritic spine density in healthy adult mouse brain. Neurosci Lett 2011; 487:260-263.
15. Dineley KT, Hogan D, Zhang WR, Taglialatela G. Acute inhibition of calcineurin restores associative learning and memory in Tg2576 APP transgenic mice. Neurobiol Learn Mem 2007; 88:217-224.
16. Shaw JA, Matlovich N, Rushlow W, Cain P, Rajakumar N. Role of calcineurin in inhibiting disadvantageous associations. Neuroscience 2012; 203:144-152.
17 .Zarrindast MR, Ardjmand A, Rezayof A, Ahmadi S. The time profile of morphine effect on different phases of inhibitory avoidance memory in rat. Arch Iran Med 2013; 16:34-37.
18. Tinsley MR, Quinn JJ, Fanselow MS. The role of muscarinic and nicotinic cholinergic neurotransmission in aversive conditioning: comparing pavlovian fear conditioning and inhibitory avoidance. Learn Mem 2004; 11:35-42.
19 .Izquierdo I, Bevilaqua LR, Rossato JI, Bonini JS, Medina JH, Cammarota M. Different molecular cascades in different sites of the brain control memory consolidation. Trends Neurosci 2006; 9:496505.
20. Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 6th ed. Amsterdam: Boston: Academic Press/Elsevier; 2007.            
21. Bennett PC, Schmidt L, Lawen A, Moutsoulas P, Ng KT. Cyclosporin A, FK506 and rapamycin produce multiple, temporally distinct, effects on memory following single-trial, passive avoidance training in the chick. Brain Res  2002; 927:180-194.
22. Ng KT, Gibbs ME, Crowe SF, Sedman GL, Hua F, Zhao W, et al. Molecular mechanisms of memory formation. Mol Neurobiol 1991;5:333-350.
23. Mansuy IM, Winder DG, Moallem TM, Osman M, Mayford M, Hawkins RD, et al. Inducible and reversible gene expression with the rtTA system for the study of memory. Neuron 1998; 21:257-265.
24. Benetoli A, Dutra AM, Paganelli RA, Senda DM, Franzin S, Milani H. Tacrolimus (FK506) reduces hippocampal damage but fails to prevent learning and memory deficits after transient, global cerebral ischemia in rats. Pharmacol Biochem Behav  2007; 88:28-38.
25. Bennett PC, Zhao W, Lawen A, Ng KT. Cyclosporin A, an inhibitor of calcineurin, impairs memory formation in day-old chicks. Brain Res 1996; 730:107-117.  
26. 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; 92:39-49.
27. Lin CH, Lee CC, Gean PW. Involvement of a calcineurin cascade in amygdala depotentiation and quenching of fear memory. Mol Pharmacol  2003; 63:44-52.
28. Lin CH, Yeh SH, Leu TH, Chang WC, Wang ST, Gean PW. Identification of calcineurin as a key signal in the extinction of fear memory. J Neurosci 2003; 23:1574-1579.
29. Christie-Fougere MM, Darby-King A, Harley CW, McLean JH. Calcineurin inhibition eliminates the normal inverted U curve, enhances acquisition and prolongs memory in a mammalian 3'-5'-cyclic AMP-dependent learning paradigm. Neuroscience 2009; 158:1277-1283.
30. Nakazawa H, Kaba H, Higuchi T, Inoue S. The importance of calmodulin in the accessory olfactory bulb in the formation of an olfactory memory in mice. Neuroscience 1995; 69:585-589.
31. Zhang JJ, Okutani F, Inoue S, Kaba H. Activation of the cyclic AMP response element-binding protein signaling pathway in the olfactory bulb is required for the acquisition of olfactory aversive learning in young rats. Neuroscience 2003; 117:707-713.
32. Zeng H, Chattarji S, Barbarosie M, Rondi-Reig L, Philpot BD, Miyakawa T, et al. Forebrain-specific calcineurin knockout selectively impairs bidirectional synaptic plasticity and working/episodic-like memory. Cell 2001; 107:617-629.
33. Mansuy IM. Calcineurin in memory and bidirectional plasticity. Biochem Biophys Res Commun 2003; 311:1195-1208.
34. Allweis C, Gibbs ME, Ng KT, Hodge RJ. Effects of hypoxia on memory consolidation: implications for a multistage model of memory. Behav Brain Res 1984; 11:117-121.
35. Sadeghnia HR, Kamkar M, Assadpour E, Boroushaki MT, Ghorbani A. Protective effect of safranal, a constituent of Crocus sativus, on quinolinic acid-induced oxidative damage in rat hippocampus. Iran J Basic Med Sci 2013; 16:73-82.
36. Sistiaga A, Sanchez-Prieto J. Protein phosphatase 2B inhibitors mimic the action of arachidonic acid and prolong the facilitation of glutamate release by group I mGlu receptors. Neuropharmacology 2000; 39:1544-1553.
37. Baldwin ML, Rostas JA, Sim AT. Two modes of exocytosis from synaptosomes are differentially regulated by protein phosphatase types 2A and 2B. J Neurochem 2003; 85:1190-1199.   
38. Volianskis A, Jensen MS. Transient and sustained types of long-term potentiation in the CA1 area of the rat hippocampus. J Physiol 2003; 550:459-492.
39. de la Fuente V, Freudenthal R, Romano A. Reconsolidation or extinction: transcription factor switch in the determination of memory course after retrieval. J Neurosci 2011; 31:5562-5573.
40. Havekes R, Nijholt IM, Visser AK, Eisel UL, Van der Zee EA. Transgenic inhibition of neuronal calcineurin activity in the forebrain facilitates fear conditioning, but inhibits the extinction of contextual fear memories. Neurobiol Learn Mem 2008; 89:595-598.
41. Torii N, Kamishita T, Otsu Y, Tsumoto T. An inhibitor for calcineurin, FK506, blocks induction of long-term depression in rat visual cortex. Neurosci Lett 1995; 185:1-4.
42 .Ikegami S, Kato A, Kudo Y, Kuno T, Ozawa F, Inokuchi K. A facilitatory effect on the induction of long-term potentiation in vivo by chronic administration of antisense oligodeoxynucleotides against catalytic subunits of calcineurin. Brain Res Mol Brain Res 1996; 41:183-191.
43. Alagarsamy S, Saugstad J, Warren L, Mansuy IM, Gereau RWt, Conn PJ. NMDA-induced potentiation of mGluR5 is mediated by activation of protein phosphatase 2B/calcineurin. Neuropharmacology 2005; 49:135-145.
44. Lu YM, Mansuy IM, Kandel ER, Roder J. Calcineurin-mediated LTD of GABAergic inhibition underlies the increased excitability of CA1 neurons associated with LTP. Neuron 2000; 26:197-205.
45 .Garthwaite J. Glutamate, nitric oxide and cell-cell signalling in the nervous system. Trends Neurosci 1991; 14:60-67.
46. .Norris CM, Blalock EM, Chen KC, Porter NM, Landfield PW. Calcineurin enhances L-type Ca(2+) channel activity in hippocampal neurons: increased effect with age in culture. Neuroscience 2002; 10:213-225.
47 .Onuma H, Lu YF, Tomizawa K, Moriwaki A, Tokuda M, Hatase O, et al. A calcineurin inhibitor, FK506, blocks voltage-gated calcium channel-dependent LTP in the hippocampus. Neurosci Res  1998; 0:313-319.
48 .Chang KT, Berg DK. Voltage-gated channels block nicotinic regulation of CREB phosphorylation and gene expression in neurons. Neuron 2001; 32:855-865.
49 .Cao W, Konsolaki M. FKBP immunophilins and Alzheimer's disease: a chaperoned affair. J Biosci 2011; 36:493-498.
50 .Kissinger CR, Parge HE, Knighton DR, Lewis CT, Pelletier LA, Tempczyk A, et al. Crystal structures of human calcineurin and the human FKBP12-FK506-calcineurin complex. Nature 1995; 78:641-644.
51 .Bito H, Deisseroth K, Tsien RW. CREB phosphorylation and dephosphorylation: a Ca(2+)- and stimulus duration-dependent switch for hippocampal gene expression. Cell 1996; 87:1203-1214.
52 .Yuan Q, Harley CW, Bruce JC, Darby-King A, McLean JH. Isoproterenol increases CREB phosphorylation and olfactory nerve-evoked potentials in normal and 5-HT-depleted olfactory bulbs in rat pups only at doses that produce odor preference learning. Learn Mem 2000; 7:413-421.
53. Azimi L, Pourmotabbed A, Ghadami MR, Nedaei SE, Pourmotabbed T. Effects of peripheral and intra-hippocampal administration of sodium salicylate on spatial learning and memory of rats. Iran J Basic Med Sci 2012; 15:709-718.
54.  Zare K, Tabatabaei SRF, Shahriari A, Jafari RA. The effect of butter oil on avoidance memory in normal and diabetic rats. Iran J Basic Med Sci 2012;
55. McLean JH, Harley CW, Darby-King A, Yuan Q. pCREB in the neonate rat olfactory bulb is selectively and transiently increased by odor preference-conditioned training. Learn Mem 1999; 6:608-618.
56. Mayford M, Bach ME, Huang YY, Wang L, Hawkins RD, Kandel ER. Control of memory formation through regulated expression of a CaMKII transgene. Science 1996; 274:1678-1683.
57.  Hakimzadeh E, Oryan S, Hajizadeh MoghadamA, Shamsizadeh A, Roohbakhsh A. Endocannabinoid system on TRPV1 receptors in the dorsal hippocampus of the rats modulate anxiety-like behaviors. Iran J Basic Med Sci  2012; 15:795–802.
58. Salehi I, Farajnia S, Mohammadi M, Sabouri GannadM. The pattern of brain-derived neurotrophic factor gene expression in the hippocampus of diabetic rats. Iran J Basic Med Sci 2010; 13:146–153.