When correlating brain size and structure with behavioural and environmental characteristics, a range of techniques can be utilised. This study used gobiid fishes to quantitatively compare brain volumes obtained via three different methods; these included the commonly used techniques of histology and approximating brain volume to an idealised ellipsoid, and the recently established technique of X-ray micro-computed tomography (micro-CT). It was found that all three methods differed significantly from one another in their volume estimates for most brain lobes. The ellipsoid method was prone to over- or under-estimation of lobe size, histology caused shrinkage in the telencephalon, and although micro-CT methods generated the most reliable results, they were also the most expensive. Despite these differences, all methods depicted quantitatively similar relationships among the four different species for each brain lobe. Thus, all methods support the same conclusions that fishes inhabiting rock pool and sandy habitats have different patterns of brain organisation. In particular, fishes from spatially complex rock pool habitats were found to have larger telencephalons, while those from simple homogenous sandy shores had a larger optic tectum. Where possible we recommend that micro-CT be used in brain volume analyses, as it allows for measurements without destruction of the brain and fast identification and quantification of individual brain lobes, and minimises many of the biases resulting from the histology and ellipsoid methods.

Keywords:
Intertidal gobies, Brain morphology, Histology, Ellipsoid model, Micro-computed tomography, Neuroecology
1.
ACMM, Sydney University (2011): X-RAY Facilities: Skyscan 1072 Micro-Computed Tomography. http://sydney.edu.au/acmm/facilities/xray/index.shtml (accessed August 14, 2011).
2.
Ayari B, Landoulsi A, Soussi-Yanicostas N (2012): Localization and characterization of kal 1.a and kal 1.b in the brain of adult zebrafish (Danio rerio). Brain Res Bull 88:345-353.
3.
Bahr GF, Bloom G, Friberg U (1957): Volume changes of tissue in physiological fluids during fixation in osmium tetroxide or formaldehyde and during subsequent treatment. Exp Cell Res 12:342-355.
4.
Bartling SH, Stiller W, Semmler W, Kiessling F (2007): Small animal computed tomography imaging. Curr Med Imaging Rev 3:45-59.
5.
Barton RA (1996): Neocortex size and behavioral ecology in primates. Proc Biol Sci 263:173-177.
6.
Braithwaite VA (2006): Cognitive ability in fish; in Sloman KA, Wilson RW, Balshine S (eds): Fish Physiology: Behaviour and Physiology of Fish. Amsterdam, Elsevier, vol 24, pp 1-37.
7.
Broglio C, Gómez A, Durán E, Salas C, Rodríguez F (2011): Brain and cognition in teleost fish; in Brown C, Laland K, Krause J (eds): Fish Cognition and Behavior, ed 2. Oxford, Wiley-Blackwell, pp 325-358.
8.
Broglio C, Rodríguez F, Salas C (2003): Spatial cognition and its neural basis in teleost fishes. Fish Fish 4:247-255.
9.
Bullmore E, Brammer M, Harvey I, Ron M (1995): Against the laterality index as a measure of cerebral asymmetry. Psychiatry Res 61:121-124.
10.
Burish MJ, Kueh HY, Wang SH (2004): Brain architecture and social complexity in modern and ancient birds. Brain Behav Evol 63:107-124.
11.
Burns JG, Rodd FH (2008): Hastiness, brain size and predation regime affect the performance of wild guppies in a spatial memory task. Anim Behav 76:911-922.
12.
Chapman AD (2009): Numbers of Living Species in Australia and the World. A Report for the Australian Biological Resources Study, Australian Biodiversity Information Services, Toowoomba, Australia. http://www.environment.gov.au/node/13866 (accessed May 24, 2014).
13.
Clayton NS (2001): Hippocampal growth and maintenance depend on food-catching experience in juvenile mountain chickadees (Poecile gambeli). Behav Neurosci 115:614-625.
14.
Costa SS, Andrade R, Carneiro LA, Gonçalves EJ, Kotrschal K, Oliveira RF (2011): Sex differences in the dorsolateral telencephalon correlate with home range size in blenniid fish. Brain Behav Evol 77:55-64.
15.
Davis RE, Northcutt RG (1983): Fish Neurobiology. Ann Arbor, University of Michigan Press.
16.
Deaner RO, Isler K, Burkart J, van Schaik C (2007): Overall brain size, and not encephalization quotient, best predicts cognitive ability across non-human primates. Brain Behav Evol 70:115-124.
17.
de Crespigny A, Bou-Reslan H, Nishimura MC, Phillips H, Carano RA, D'Arceuil HE (2008): 3D micro-CT imaging of the postmortem brain. J Neurosci Methods 171:207-213.
18.
Degenhardt K, Wright AC, Horng D, Padmanabhan A, Epstein JA (2010): Rapid 3D phenotyping of cardiovascular development in mouse embryos by micro-CT with iodine staining. Circ Cardiovasc Imaging 3:314-322.
19.
Dobrivojević M, Bohaček I, Erjavec I, Gorup D, Gajović S (2013): Computed microtomography visualization and quantification of mouse ischemic brain lesion by nonionic radio contrast agents. Croat Med J 54:3-11.
20.
Dunbar RIM (1995): Neocortex size and group size in primates: a test of the hypothesis. J Hum Evol 28:287-296.
21.
Garamszegi LZ, Eens M, Erritzø J, Møller AP (2005): Sperm competition and sexually size dimorphic brains in birds. Proc Biol Sci 272:159-166.
22.
Garcia-Finana M, Cruz-Orive LM, Mackay CE, Pakkenberg B, Roberts N (2003): Comparison of MR imaging against physical sectioning to estimate the volume of human cerebral compartments. Neuroimage 18:505-516.
23.
Gonda A, Herczeg G, Merilä J (2009): Habitat-dependent and -independent plastic responses to social environment in the nine-spined stickleback (Pungitius pungitius) brain. Proc Biol Sci 276:161-167.
24.
Gonzalez-Voyer A, Winberg S, Kolm N (2009a): Social fishes and single mothers: brain evolution in African cichlids. Proc Biol Sci 276:161-167.
25.
Gonzalez-Voyer A, Winberg S, Kolm N (2009b): Brain structure evolution in a basal vertebrate clade: evidence from phylogenetic comparative analysis of cichlid fishes. BMC Evol Biol 9:238.
26.
Gonzalez-Voyer A, Kolm N (2010): Sex, ecology and the brain: evolutionary correlates of brain structure volumes in Tanganyikan cichlids. PLoS One 5:e14355.
27.
Grace AA, Llinas R (1985): Morphological artifacts induced in intracellularly stained neurons by dehydration: circumvention using rapid dimethyl sulfoxide clearing. Neuroscience 16:461-475.
28.
Haug H (1986): History of neuromorphometry. J Neurosci Methods 12:1-17.
29.
Hayasaka N, Nagai N, Kawao N, Niwa A, Yoshioka Y, et al. (2012): In vivo diagnostic imaging using micro-CT: sequential and comparative evaluation of rodent models for hepatic/brain ischemia and stroke. PLoS One 7:e32342.
30.
Healy S, Guilford T (1990): Olfactory-bulb size and nocturnality in birds. Evolution 44:339-346.
31.
Healy SD, Rowe C (2006): A critique of comparative studies of brain size. Proc Biol Sci 274:453-464.
32.
Hsieh J (2003): Computed Tomography - Principles, Design, Artifacts, and Recent Advances. Bellingham, SPIE Press.
33.
Huber R, Van Staaden MJ, Kaufman LS, Liem KF (1997): Microhabitat use, trophic patterns, and the evolution of brain structure in African Cichlids. Brain Behav Evol 50:167-182.
34.
Hutcheon JM, Kirsch JW, Garland T (2002): A comparative analysis of brain size in relation to foraging ecology and phylogeny in the chiroptera. Brain Behav Evol 60:165-180.
35.
Iwaniuk AN, Nelson JE (2001): A comparative analysis of relative brain size in waterfowl (Anseriformes). Brain Behav Evol 57:87-97.
36.
Iwaniuk AN, Nelson JE (2002): Can endocranial volume be used as an estimate of brain size in birds. Can J Zool 80:16-23.
37.
Jelsing J, Rostrup E, Markenroth K, Paulson OB, Gundersen HJG, Hemmingsen R, Pakkenberg B (2005): Assessment of in vivo MR imaging compared to physical sections in vitro: a quantitative study of brain volumes using stereology. Neuroimage 26:57-65.
38.
Johnson JT, Hansen MS, Wu I, Healy LJ, Johnson CR, Jones GM, Capecchi MR, Keller C (2006): Virtual histology of transgenic mouse embryos for high-throughput phenotyping. PLoS Genet 2:e61.
39.
Kalender WA (2005): Computed Tomography: Fundamentals, System Technology, Image Quality, Applications. Erlangen, Publicis Corporate Publishing.
40.
Kihslinger RL, Lema SC, Nevitt GA (2006): Environmental rearing conditions produce forebrain differences in wild Chinook salmon Oncorhynchus tshawytscha. Comp Biochem Physiol A Mol Integr Physiol 145:145-151.
41.
Kihslinger RL, Nevitt A (2006): Early rearing environment impacts cerebellar growth in juvenile salmon. J Exp Biol 209:504-509.
42.
Kim TH, Zollinger L, Shi XF, Rose J, Jeong EK (2009): Diffusion tensor imaging of ex vivo cervical spinal cord specimens: the immediate and long-term effects of fixation on diffusivity. Anat Rec (Hoboken) 292:234-241.
43.
Kolm N, Gonzalez-Voyer A, Brelin D, Winberg S (2009): Evidence for small scale variation in the vertebrate brain: mating strategy and sex affect brain size and structure in wild brown trout (Salmo trutta). J Evol Biol 22:2524-2531.
44.
Kotrschal K, Junger H (1988): Patterns of brain morphology in mid-European Cyprinidae (Pisces, Teleostei): a quantitative histological study. J Hirnforsch 29:341-352.
45.
Kotrschal K, Palzenberger M (1992): Neuroecology of cyprinids: comparative, quantitative histology reveals diverse brain patterns. Environ Biol Fishes 33:135-152.
46.
Kotrschal, K. Van Staaden MJ, Huber R (1998): Fish brains: evolution and environmental relationships. Rev Fish Biol Fisher 8:373-408.
47.
Kotrschal A, Sundstrom LF, Brelin D, Devlin RH, Kolm N (2012a): Inside the heads of David and Goliath: environmental effects on brain morphology among wild and growth-enhanced coho salmon Oncorhynchus kisutch. J Fish Biol 81:987-1002.
48.
Kotrschal A, Räsänen K, Kristjánsson BK, Senn M, Kolm N (2012b): Extreme sexual brain size dimorphism in sticklebacks: a consequence of the cognitive challenges of sex and parenting? PLoS One 7:e30055.
49.
LaDage LD, Roth TC 2nd, Pravosudov VV (2009): Biases in measuring the brain: the trouble with the telencephalon. Brain Behav Evol 73:253-258.
50.
Lecchini D, Lecellier G, Lanyon RG, Holles S, Poucet B, Duran E (2014): Variation in brain organization of coral reef fish larvae according to life history traits. Brain Behav Evol 83:17-30.
51.
Lisney TJ, Collin SP (2006): Brain morphology in large pelagic fishes: a comparison between sharks and teleosts. J Fish Biol 68:532-554.
52.
Lisney TJ, Bennett MB, Collin SP (2007): Volumetric analysis of sensory brain areas indicates ontogenetic shifts in the relative importance of sensory systems in elasmobranchs. Raffles Bull Zool 14:7-15.
53.
Metscher BD (2009a): MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiol 9:11.
54.
Metscher BD (2009b): MicroCT for developmental biology: a versatile tool for high-contrast 3D imaging at histological resolutions. Dev Dyn 238:632- 640.
55.
Mizutani R, Takeuchi A, Hara T, Uesugi K, Suzuki Y (2007): Computed tomography imaging of the neuronal structure of Drosophila brain. J Synchrotron Radiat 14:282-287.
56.
Neues F, Epple M (2008): X-ray microcomputer tomography for the study of biomineralized endo- and exoskeletons of animals. Chem Rev 108:4734-4741.
57.
Nieuwenhuys R, Meek J (1998): Holosteans and teleosts; in Nieuwenhuys R, Ten Donkelaar HJ, Nicholoson C (eds): The Central Nervous System of Vertebrates. Berlin, Springer, vol 2, pp 759-938.
58.
Paterson GLJ, Sykes D, Faulwetter S, Merk R, Ahmed F, Hawkins LE, Dinley J, Ball AD, Arvanitidis C (2014): The pros and cons of using micro-computed tomography in gross and microanatomical assessments of polychaetous annelids. Memoirs Museum Victoria 71:237-246. from http://museumvictoria.com.au/about/books-and-journals/journals/memoirs-of-museum-victoria/ (accessed February 9, 2015).
59.
Pitnick S, Jones KE, Wilkinson GS (2006): Mating system and brain size in bats. Proc Biol Sci 273:719-724.
60.
Pollen AA, Dobberfuhl AP, Scace J, Igulu MM, Renn SCP, Shumway CA, Hofmann HA (2007): Environmental complexity and social organization sculpt the brain in Lake Tanganyikan cichlid fish. Brain Behav Evol 70:21-39.
61.
Prajapati SI, Kilcoyne A, Samano AK, Green DP, McCarthy SD, Blackman BA, Brady MM, Zarzabal LA, Tatiparthy AK, Sledz TJ, Duong T, Ohshima-Hosoyama S, Giles FJ, Michalek JE, Rubin BP, Keller C (2011): MicroCT-based virtual histology evaluation of preclinical medulloblastoma. Mol Imaging Biol 13:493-499.
62.
Purea A, Webb AG (2006): Reversible and irreversible effects of chemical fixation on the NMR properties of single cells. Magn Reson Med 56:927-931.
63.
Reader SM, Laland KN (2002): Social intelligence, innovation, and enhanced brain size in primates. Proc Natl Acad Sci USA 99:4436-4441.
64.
Ribi W, Senden TJ, Sakellariou A, Limaye A, Zhang S (2008): Imaging honey bee brain anatomy with micro-X-ray-computed tomography. J Neurosci Methods 171:93-97.
65.
Rosen GD, Harry JD (1990): Brain volume estimation from serial section measurements: a comparison of methodologies. J Neurosci Methods 35:115-124.
66.
Safi K, Dechmann DK (2005): Adaptation of brain regions to habitat complexity: a comparative analysis in bats (Chiroptera). Proc Biol Sci 272:179-186.
67.
Salas C, Broglio C, Rodriguez F, Lopez JC, Portavella M, Torres B (1996): Telencephalic ablation in goldfish impairs performance in a ‘spatial constancy' problem but not in a cued one. Behav Brain Res 79:193-200.
68.
Schambach SJ, Bag S, Schilling L, Groden C, Brockmann MA (2010): Application of micro-CT in small animal imaging. Methods 50:2-13.
69.
Shultz S, Dunbar RI (2006): Both social and ecological factors predict ungulate brain size. Proc Biol Sci 273:207-215.
70.
Stowell RE (1941): Effect on tissue volume of various methods of fixation, dehydration, and embedding. Biotech Histochem 16:67-83.
71.
Striedter GF (2005): Principles of Brain Evolution. Sunderland, Sinauer Associates.
72.
Ullmann JFP, Cowin G, Collin SP (2010): Quantitive assessment of brain volumes in fish: comparison of methodologies. Brain Behav Evol 76:261-270.
73.
Uylings HBM, van Eden CG, Hofman MA (1986): Morphometry of size/volume variables and comparison of their bivariate relations in the nervous system under different conditions. J Neurosci Methods 18:19-37.
74.
Van Staaden MJ, Huber R, Kaufman LS, Liem KF (1995): Brain evolution in cichlids of the African Great Lakes: brain and body size, general patterns, and evolutionary trends. Zoology 98:165-178.
75.
Vargas JP, Rodriguez F, Lopez JC, Arias JL, Salas C (2000): Spatial learning-induced increase in the argyrophilic nucleolar organizer region of dorsolateral telencephalic neurons in goldfish. Brain Res 865:77-84.
76.
Wagner HJ (2001a): Sensory brain areas in mesopelagic fishes. Brain Behav Evol 57:117-133.
77.
Wagner HJ (2001b): Brain areas in abyssal demersal fishes. Brain Behav Evol 27:301-316.
78.
Weil A (1928): The measurement of cerebral and cerebellar surfaces. V. The determination of the shrinkage of the surface of different vertebrate brains. Arch Neurol Psychiatry 20:834-835.
79.
White GE (2015): Microhabitat use affects brain size and structure in intertidal gobies. Brain Behav Evol, Epub ahead of print.
80.
Wilson ADM, McLaughlin RL (2010): Foraging behaviour and brain morphology in recently emerged brook charr, Salvelinus fontinalis. Behav Ecol Sociobiol 64:1905-1914.
81.
Wiper ML, Britton S, Higgs DM (2014): Early experience and reproductive morph both affect brain morphology in adult male Chinook salmon (Oncorhynchus tshawytscha). Can J Fish Aquat Sci 71:1430-1436.
82.
Wullimann MF, Rupp B, Reichert H (1996): Neuroanatomy of the Zebrafish Brain: A Topological Atlas. Basel, Birkhäuser.
83.
Wullimann MF, Mueller T (2004): Teleostean and mammalian forebrain contrasted: evidence from genes to behavior. J Comp Neurol 475:143-162.
84.
Young LJ, Wang Z, Insel TR (1998): Neuroendocrine bases of monogamy. Trends Neurosci 21:71-75.
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