Predation pressure represents a strong selective force that influences the development and evolution of living organisms. An increasing number of studies have shown that both environmental and social factors, including exposure to predators, substantially shape the structure and function of the brain. However, our knowledge about the molecular mechanisms underlying the response of the brain to environmental stimuli is limited. In this study, we used whole-genome comparative oligonucleotide microarrays to investigate the brain transcriptomic response to cues of a predator in the threespine stickleback, Gasterosteus aculeatus. We found that repeated exposure to olfactory, visual and tactile cues of a predator (rainbow trout, Oncorrhynchus mykiss) for 6 days resulted in subtle but significant transcriptomic changes in the brain of sticklebacks. Gene functional analysis and gene ontology enrichment revealed that the majority of the transcripts differentially expressed between the fish exposed to cues of a predator and the control group were related to antigen processing and presentation involving the major histocompatibility complex, transmission of synaptic signals, brain metabolic processes, gene regulation and visual perception. The top four identified pathways were synaptic long-term depression, RAN signaling, relaxin signaling and phototransduction. Our study demonstrates that exposure of sticklebacks to cues of a predator results in the activation of a wide range of biological and molecular processes and lays the foundation for future investigations on the molecular factors that modulate the function and evolution of the brain in response to stressors.

1.
Apanius V, Penn D, Slev PR, Ruff LR, Potts WK (1997): The nature of selection on the major histocompatibility complex. Crit Rev Immunol 17:179–224.
2.
Baker JA, Foster SA, Heins DC, Bell MA, King RW (1998): Variation in female life-history traits among Alaskan populations of the threespine stickleback, Gasterosteus aculeatus L. (Pisces: Gasterosteidae). Biol J Linn Soc Lond 63:141–159.
3.
Bear MF (1999): Homosynaptic long-term depression: a mechanism for memory? Proc Natl Acad Sci USA 96:9457–9458.
4.
Bell AM (2005): Behavioural differences between individuals and two populations of stickleback (Gasterosteus aculeatus). J Evol Biol 18:464–473.
5.
Bell AM, Backstrom T, Huntingford FA, Pottinger TG, Winberg S (2007): Variable neuroendocrine responses to ecologically-relevant challenges in sticklebacks. Physiol Behav 91:15–25.
6.
Bell AM, Dingemanse NJ, Hankison S, Langenhof M, Rollins, K (2011): Early exposure to nonlethal predation risk by size-selective predators increases somatic growth and decreases size at adulthood in threespined sticklebacks. J Evol Biol 24:943–945.
7.
Bell AM, Henderson L, Huntingford FA (2010): Behavioral and respiratory responses to stressors in multiple populations of three-spined sticklebacks that differ in predation pressure. J Comp Physiol B 180:211–220.
8.
Bell AM, Sih A (2007): Exposure to predation generates personality in threespined sticklebacks. Ecol Lett10:828–834.
9.
Bell MA, Foster SA (1994): Introduction to the evolutionary biology of the threespine stickleback; in Bell MA, Foster SA (eds): The Evolutionary Biology of the Threespine Stickleback. Oxford, Oxford University Press, pp 1–27.
10.
Benjamini Y, Hochberg Y (1995): Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B (Methodological) 57:289–300.
11.
Bernatchez L, Landry C (2003): MHC studies in nonmodel vertebrates: what have we learned about natural selection in 15 years? J Evol Biol 16:363–377.
12.
Blanchard DC, Canteras NS, Markham CM, Pentkowski NS, Blanchard RJ (2005): Lesions of structures showing FOS expression to cat presentation: effects on responsivity to a cat, cat odor, and nonpredator threat. Neurosci Biobehav Rev 29:1243–1253.
13.
Bourgon R, Gentleman R, Huber W (2010): Independent filtering increases detection power for high-throughput experiments. Proc Natl Acad Sci USA 107:9546–9551.
14.
Brown GE, Godin JJ (1997): Anti-predator responses to conspecific and heterospecific skin extracts by threespine sticklebacks: alarm pheromones revisited. Behaviour 134:1123–1134.
15.
Brown MM, Williams TD, Chipman JK, Katsiadaki I, Sanders M, Craft JA (2008): Construction of subtracted EST and normalised cDNA libraries from liver of chemical-exposed three-spined stickleback (Gasterosteus aculeatus) containing pollutant-responsive genes as a resource for transcriptome analysis. Mar Env Res 66:127–130.
16.
Cairns MT, Johnson MC, Talbot AT, Pernmasani JK, McNeill RE, Houeix B, Sangrador-Vegas A, Pottinger, TG (2008): A cDNA microarray assessment of gene expression in the liver of rainbow trout (Oncorhynchus mykiss) in response to a handling and confinement stressor. Comp Biochem Physiol D Genomics Proteomics 3:51–66.
17.
Calduch-Giner JA, Davey G, Saera-Vila A, Houeix B, Talbot A, Prunet P, Cairns MT, Perez-Sanchez J (2010): Use of microarray technology to assess the time course of liver stress response after confinement exposure in gilthead sea bream (Sparus aurata L.). BMC Genomics 11:No. 193.
18.
Calza S. Raffelsberger W, Ploner A, Sahel J, Leveillard T, Pawitan J (2007): Filtering genes to improve sensitivity in oligonucleotide microarray data analysis. Nucleic Acids Res 35:e102.
19.
Cantor C (2009): Post-traumatic stress disorder: evolutionary perspectives. Aust NZJ Psychiatry 43:1038–1048.
20.
Carvalho B, Bengtsson H, Speed TP, Irizarry RA (2007): Exploration, normalization, and genotype calls of high-density oligonucleotide SNP array data. Biostatistics 8:485–499.
21.
Chan YF, Marks ME, Jones FC, Villarreal G Jr, Shapiro MD, Brady SD, Southwick AM, Absher DM, Grimwood J, Schmutz J, Myers RM, Petrov D, Jonsson B, Schluter D, Bell A, Kingsley DM (2010): Adaptive evolution of pelvic reduction in sticklebacks by recurrent deletion of a Pitx1 enhancer. Science 327:302–305.
22.
Ching B, Jamieson S, Heath JW, Heath DD, Hubberstey A (2010): Transcriptional differences between triploid and diploid Chinook salmon (Oncorhynchus tshawytscha) during live Vibrio anguillarum challenge. Heredity 104:224–234.
23.
Chivers DP, Wisenden BD, Hindman CJ, Michalak TA, Kusch RC, Kaminskyj SG, Jack KL, Ferrari MC, Pollock RJ, Halbgewachs CF, Pollock MS, Alemadi S, James CT, Savaloja RK, Goater CP, Corwin A, Mirza RS, Kiesecker JM, Brown GE, Adrian JC Jr, Krone PH, Blaustein AR, Mathis A (2007): Epidermal ‘alarm substance’ cells of fishes maintained by non-alarm functions: possible defence against pathogens, parasites and UVB radiation. Proc Biol Sci 274:2611–2619.
24.
Cocke R, Moynihan JA, Cohen N, Grota LJ, Ader R (1993): Exposure to conspecific alarm chemosignals alters immune responses in BALB/c mice. Brain Behav Immun 7:36–46.
25.
Colosimo PF, Hosemann KE, Balabhadra S, Villarreal G Jr, Dickson M, Grimwood J, Schmutz J, Myers RM, Schluter D, Kingsley DM (2005): Widespread parallel evolution in sticklebacks by repeated fixation of ectodysplasin alleles. Science 307:1928–1933.
26.
Comoli E, Ribeiro-Barbosa ER, Canteras NS (2003): Predatory hunting and exposure to a live predator induce opposite patterns of Fos immunoreactivity in the PAG. Behav Brain Res 138:17–28.
27.
Corriveau RA, Huh GS, Shatz CJ (1998): Regulation of class I MHC gene expression in the developing and mature CNS by neural activity. Neuron 21:505–520.
28.
Craig P M, Hogstrand C, Wood CM, McClelland GB (2009): Gene expression endpoints following chronic waterborne copper exposure in a genomic model organism, the zebrafish, Danio rerio. Physiol Genomics 40:23–33.
29.
Cresko WA, Amores A, Wilson C, Murphy J, Currey M, Phillips P, Bell MA, Kimmel CB, Postlethwait JH (2004): Parallel genetic basis for repeated evolution of armor loss in Alaskan threespine stickleback population. Proc Nat Acad Sci 101:6050–6055.
30.
Diamond DM, Park CR (2000): Predator exposure produces retrograde amnesia and blocks synaptic plasticity. Progress toward understanding how the hippocampus is affected by stress. Ann NY Acad Sci 911:453–455.
31.
Dierick HA, Greenspan J (2006): Molecular analysis of flies selected for aggressive behavior. Nat Genet 38:1023–1031.
32.
Dingemanse NJ, Van der Plas F, Wright J, Reale D, Schrama M, Roff DA, Van der Zee E, Barber I (2009): Individual experience and evolutionary history of predation affect expression of heritable variation in fish personality and morphology. Proc Biol Sci 276:1285–1293.
33.
Egid K, Brown JL (1989): The major histocompatibility complex and female mating preference in mice. Anim Behav 38:548–549.
34.
Feldman AJ, Costouros NG, Wang E, Qian M, Marincola FM, Alexander HR, Libutti SK (2002): Advantages of mRNA amplification for microarray analysis. Biotechniques33:906–914.
35.
Garrett SH, Somji S, Sens MA, Zhang K, Sens DA (2011): Microarray analysis of gene expression patterns in human proximal tubule cells over a short and long time course of cadmium exposure. J Toxicol Environ Health 74:24–42.
36.
Geoghegan F, Katsiadaki I, Williams, TD, Chipman JK (2008): A cDNA microarray for the three-spined stickleback, Gasterosteus aculeatus L., and analysis of the interactive effects of oestradiol and dibenzanthracene exposures. J Fish Biol 72:2133–2153.
37.
Giles N, Huntingford FA (1984): Predation risk and inter-population variation in antipredator behaviour in the threespined stickleback. Anim Behav 32:264–275.
38.
Goddard CA, Butts DA, Shatz CJ (2007): Regulation of CNS synapses by neuronal MHC class I. Proc Natl Acad Sci 104:6828–6833.
39.
Holderbach R, Clark K, Moreau JL, Bischofberger J, Normann C (2007): Enhanced long-term synaptic depression in an animal model of depression. Biol Psychiatry 62:92–100.
40.
Huntingford FA, Ruiz-Gomez ML (2009): Three-spined sticklebacks Gasterosteus aculeatus as a model for exploring behavioural biology. J Fish Biol 75:1943–1976.
41.
Huntingford FA, Wright PJ, Tierney JF (1994): Adaptive variation and antipredator behaviour in threespine stickleback; in Bell MA, Foster SA (eds): The Evolutionary Biology of the Threespine Stickleback.Oxford, Oxford University Press, pp 277–295.
42.
Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003): Summaries of affymetrix GeneChip probe level data. Nucleic Acids Res 31:e15.
43.
Kelley JL, Magurran AE (2003): Learned predator recognition and antipredator responses in fishes. Fish Fish 4:216–226.
44.
Kennerly E, Ballmann A, Martin S, Wolfinger R, Gregory S, Stoskopf M, Gibson G (2008): A gene expression signature of confinement in peripheral blood of red wolves (Canis rufus). Mol Ecol 17:2782–2791.
45.
Kim JJ, Diamond D (2002): The stressed hippocampus, synaptic plasticity and lost memories. Nat Rev Neurosci 3:453–462.
46.
Kingsley DM, Peichel CL (2007): The molecular genetics of evolutionary change in sticklebacks; in Östlund-Nilsson O, Mayer I, Huntingford FA (eds): Biology of the Three-Spined Stickleback. Boca Raton, CRC Press.
47.
Krasnov A, Koskinen H, Pehkonen P, Rexroad CE, Afanasyev S, Molsa H (2005): Gene expression in the brain and kidney of rainbow trout in response to handling stress. BMC Genomics 6:3.
48.
Kuzmiski JB, Marty V, Baimoukhametova DV, Bains JS (2010). Stress-induced priming of glutamate synapses unmasks associative short-term plasticity. Nat Neurosci 13:1257–1264.
49.
Leder EH, Merila J, Primmer CR (2009): A flexible whole-genome microarray for transcriptomics in three-spine stickleback (Gasterosteus aculeatus). BMC Genomics 10:426.
50.
Magurran AE (1990): The inheritance and development of minnow antipredator behavior. Anim Behav 39:834–842.
51.
Malleret G, Alarcon JM, Martel G, Takizawa, S, Vronskaya S, Yin D, Chen IZ, Kandel ER, Shumyatsky GP (2010): Bidirectional regulation of hippocampal long-term synaptic plasticity and its influence on opposing forms of memory. J Neurosci 30:3813–3825.
52.
Marchinko KB (2009): Predation’s role in repeated phenotypic and genetic divergence of armor in threespine stickleback. Evolution 63:127–138.
53.
Masuda T, Iigo M, Mizusawa K, Aida K (2003): Retina-type rhodopsin gene expressed in the brain of a teleost, ayu (Plecoglossus altivelis). Zoolog Sci 20:989–997.
54.
McEwen BS (2007): Physiology and neurobiology of stress and adaptation: central role of the brain. Physiol Rev 87:873–904.
55.
Milinski M, Griffiths S, Wegner KM, Reusch TB, Haas-Assenbaum A, Boehm T (2005): Mate choice decisions of stickleback females predictably modified by MHC peptide ligands. Proc Natl Acad Sci 102:4414–4418.
56.
Moodie GEE (1972): Predation, natural selection and adaptation in an unusual threespine stickleback. Evolution 28:167.
57.
Mori T, Kawachi H, Imai C, Sugiyama M, Kurata Y, Kishida O, Nishimura K (2009): Identification of a novel uromodulin-like gene related to predator-induced bulgy morph in anuran tadpoles by functional microarray analysis. PLoS One 4:e5936.
58.
Motta SC, Goto M, Gouveia FV, Baldo MV, Canteras NS, Swanson LW (2009): Dissecting the brain’s fear system reveals the hypothalamus is critical for responding in subordinate conspecific intruders. Proc Natl Acad Sci 106:4870–4875.
59.
Mozhui K, Ciobanu DC, Schikorski T, Wang X, Lu L, Williams RW (2008): Dissection of a QTL hotspot on mouse distal chromosome 1 that modulates neurobehavioral phenotypes and gene expression. PLoS Genet 4:e1000260.
60.
Mujica-Parodi LR, Strey HH, Frederick B, Savoy R, Cox D, Botanov Y, Tolkunov D, Rubin D, Weber J (2009): Chemosensory cues to conspecific emotional stress activate amygdala in humans. PLoS One 4:e6415.
61.
Mukai M, Replogle K, Drnevich J, Wang G, Wacker D, Band M, Clayton DF, Wingfield JC (2009): Seasonal differences of gene expression profiles in song sparrow (Melospiza melodia) hypothalamus in relation to territorial aggression. PLoS One 4:e8182.
62.
Munoz-Sanjuan I, Brivanlou AH (2005): Induction of ectopic olfactory structures and bone morphogenetic protein inhibition by Rossy, a group XII secreted phospholipase A2. Mol Cell Biol 25:3608–3619.
63.
Nanda SA, Qi C, Roseboom PH, Kalin NH (2008): Predator stress induces behavioral inhibition and amygdala somatostatin receptor 2 gene expression. Genes Brain Behav 7:639–648.
64.
Oliveira AL, Thams S, Lidman O, Piehl F, Hokfelt T, Karre K, Linda H, Cullheim, S (2004): A role for MHC class I molecules in synaptic plasticity and regeneration of neurons after axotomy. Proc Natl Acad Sci 101:17843–17848.
65.
Olsen KH, Grahn M, Lohm J, Langefors A (1998): MHC and kin discrimination in juvenile arctic charr, Salvelinus alpinus (L.). Anim Behav 56:319–327.
66.
Peichel CL, Nereng KS, Ohgi KA, Cole BL, Colosimo PF, Buerkle CA, Schluter D, Kingsley, DM (2001): The genetic architecture of divergence between threespine stickleback species. Nature 414:901–905.
67.
Pfaffl MW, Horgan GW, Dempfle L (2002): Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:e36.
68.
Polacek DC, Passerini AG, Shi C, Francesco NM, Manduchi E, Grant GR, Powell S, Bischof H, Winkler H, Stoeckert CJ Jr, Davies PF (2003): Fidelity of enhanced sensitivity of differential transcription profiles following linear amplification of nanogram amounts of endothelial mRNA. Physiol Genomics13:147–156.
69.
Ponder CA, Huded CP, Munoz MB, Gulden FO, Gilliam TC, Palmer AA (2008): Rapid selection response for contextual fear conditioning in a cross between C57BL/6J and A/J: behavioral, QTL and gene expression analysis. Behav Genet 38:277–291.
70.
R Development Core Team (2009): R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna. ISBN 3-900051-07-0, URLhttp://www.R-Project.Org.
71.
Reimchen TE (1980): Spine deficiency and polymorphism in a population of Gasterosteus aculeatus – an adaptation to predators. Can J Zool 58:1232–124.
72.
Reimchen TE (1994):Predators and morphological evolution in threespine stickleback;in Bell MA, Foster SA (eds): The Evolutionary Biology of the Threespine Stickleback. Oxford, Oxford University Press, pp 240–273.
73.
Roseboom PH, Nanda, SA, Bakshi VP, Trentani A, Newman SM, Kalin NH (2007): Predator threat induces behavioral inhibition, pituitary-adrenal activation and changes in amygdala CRF-binding protein gene expression. Psychoneuroendocrinology 32:44–55.
74.
Shapiro MD, Marks ME, Peichel CL, Blackman BK, Nereng KS, Jonsson B, Schluter D, Kingsley, DM (2004): Genetic and developmental basis of evolutionary pelvic reduction in threespine sticklebacks. Nature 428:717–723.
75.
Shatz CJ (2009): MHC class I: an unexpected role in neuronal plasticity. Neuron 64:40–45.
76.
Shiina T, Ota M, Shimizu S, Katsuyama Y, et al (2006): Rapid evolution of major histocompatibility complex class I genes in primates generates new disease alleles in humans via hitchhiking diversity. Genetics 173:1555–1570.
77.
Spurgin LG, Richardson DS (2010): How pathogens drive genetic diversity: MHC, mechanisms and misunderstandings. Proc Biol Sci 277:979–988.
78.
Tulley TJ, Huntingford FA (1987): Paternal care and the development of adaptive variation in anti-predator responses in sticklebacks. Anim Behav 35:1570–1572.
79.
Wang H, Zhu YZ, Wong PT, Farook JM, Teo AL, Lee LK, Moochhala S (2003): cDNA microarray analysis of gene expression in anxious PVG and SD rats after cat-freezing test. Exp Brain Res 149:413–421.
80.
Wedekind C, Seebeck T, Bettens F, Paepke AJ (1995): MHC-dependent mate preferences in humans. Proc Biol Sci 260:245–249.
81.
Williams TD, Brown M, Chipman JK, et al (2008): Development of a stickleback (Gasterosteus aculeatus) cDNA microarray and gene expression responses to dibenzanthracene, ethinyl-estradiol and copper. Marine Environ Res 66:140–140.
82.
Wootton RJ (1976): The Biology of the Stickleback. London, Academic Press.
83.
Yang Y, Zheng X, Wang Y, Cao J, Dong Z, Cai J, Sui N, Xu L (2004): Stress enables synaptic depression in CA1 synapses by acoustic and chronic morphine: Possible mechanisms for corticosterone on opiate addiction. J Neurosci 24:2412–2420.
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