Abstract
Closely related substrains of inbred mice often show phenotypic differences that are presumed to be caused by recent mutations. The substrains BALB/cJ and BALB/cByJ, which were separated in 1935, have been reported to show numerous highly significant behavioral and morphological differences. In an effort to identify some of the causal mutations, we phenotyped BALB/cJ and BALB/cByJ mice as well as their F1, F2, and N2 progeny for behavioral and morphological phenotypes. We also generated whole-genome sequence data for both inbred strains (~3.5× coverage) with the intention of identifying polymorphic markers to be used for linkage analysis. We observed significant differences in body weight, the weight of the heart, liver, spleen and brain, and corpus callosum length between the two substrains. We also observed that BALB/cJ animals showed greater anxiety-like behavior in the open field test, less depression-like behavior in the tail suspension test, and reduced aggression compared to BALB/cByJ mice. Some but not all of these physiological and behavioral results were inconsistent with prior publications. These inconsistencies led us to suspect that the differences were due to, or modified by, non-genetic factors. Thus, we did not perform linkage analysis. We provide a comprehensive summary of the prior literature about phenotypic differences between these substrains as well as our current findings. We conclude that many differences between these strains are unstable and therefore ill-suited to linkage analysis; the source of this instability is unclear. We discuss the broader implications of these observations for the design of future studies.
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References
Blizard DA, Vandenbergh DJ, Jefferson AL et al (2004) Effects of periadolescent ethanol exposure on alcohol preference in two BALB substrains. Alcohol 34:177–185. doi:10.1016/j.alcohol.2004.08.007
Bohlen MO, Bailoo JD, Jordan RL, Wahlsten D (2012) Hippocampal commissure defects in crosses of four inbred mouse strains with absent corpus callosum. Genes Brain Behav 11:757–766. doi:10.1111/j.1601-183X.2012.00802.x
Bryant CD, Zhang NN, Sokoloff G et al (2008) Behavioral differences among C57BL/6 substrains: implications for transgenic and knockout studies. J Neurogenet 22:315–331. doi:10.1080/01677060802357388
Bryant CD, Graham ME, Distler MG et al (2009) A role for casein kinase 1 epsilon in the locomotor stimulant response to methamphetamine. Psychopharmacology 203:703–711. doi:10.1007/s00213-008-1417-z
Bull KR, Rimmer AJ, Siggs OM et al (2013) Unlocking the bottleneck in forward genetics using whole-genome sequencing and identity by descent to isolate causative mutations. PLoS Genet. doi:10.1371/journal.pgen.1003219
Chesler EJ, Wilson SG, Lariviere WR et al (2002) Identification and ranking of genetic and laboratory environment factors influencing a behavioral trait, thermal nociception, via computational analysis of a large data archive. Neurosci Biobehav Rev 26:907–923. doi:10.1016/S0149-7634(02)00103-3
Ciaranello RD, Lipsky A, Axelrod J (1974) Association between fighting behavior and catecholamine biosynthetic enzyme activity in two inbred mouse sublines. Proc Natl Acad Sci U S A 71:3006–3008. doi:10.1073/pnas.71.8.3006
Cook MN, Bolivar VJ, McFadyen MP, Flaherty L (2002) Behavioral differences among 129 substrains: implications for knockout and transgenic mice. Behav Neurosci 116:600–611. doi:10.1037/0735-7044.116.5.756
Couppis MH, Kennedy CH, Stanwood GD (2008) Differences in aggressive behavior and in the mesocorticolimbic DA system between A/J and BALB/cJ mice. Synapse 62:715–724. doi:10.1002/syn.20545
Crabbe JC, Wahlsten D, Dudek BC (1999) Genetics of mouse behavior: interactions with laboratory environment. Science 284:1670–1672. doi:10.1126/science.284.5420.1670
Crowley JJ, Blendy JA, Lucki I (2005) Strain-dependent antidepressant-like effects of citalopram in the mouse tail suspension test. Psychopharmacology 183:257–264. doi:10.1007/s00213-005-0166-5
Crowley JJ, Brodkin ES, Blendy JA et al (2006) Pharmacogenomic evaluation of the antidepressant citalopram in the mouse tail suspension test. Neuropsychopharmacology 31:2433–2442. doi:10.1038/sj.npp.1301065
Dow HC, Kreibich AS, Kaercher KA et al (2011) Genetic dissection of intermale aggressive behavior in BALB/cJ and A/J mice. Genes Brain Behav 10:57–68. doi:10.1111/j.1601-183X.2010.00640.x
Dulawa SC, Holick KA, Gundersen B, Hen R (2004) Effects of chronic fluoxetine in animal models of anxiety and depression. Neuropsychopharmacology 29:1321–1330. doi:10.1038/sj.npp.1300433
Englander MT, Dulawa SC, Bhansali P, Schmauss C (2005) How stress and fluoxetine modulate serotonin 2C receptor pre-mRNA editing. J Neurosci 25:648–651. doi:10.1523/JNEUROSCI.3895-04.2005
Eppig JT, Blake J a, Bult CJ et al (2012) The Mouse Genome Database (MGD): comprehensive resource for genetics and genomics of the laboratory mouse. Nucleic Acids Res 40:D881–D886. doi:10.1093/nar/gkr974
Fairless AH, Dow HC, Toledo MM et al (2008) Low sociability is associated with reduced size of the corpus callosum in the BALB/cJ inbred mouse strain. Brain Res 1230:211–217. doi:10.1016/j.brainres.2008.07.025
Fonio E, Golani I, Benjamini Y (2012) Measuring behavior of animal models: faults and remedies. Nat Methods 9:1167–1170. doi:10.1038/nmeth.2252
Hilakivi LA, Lister RG (1989) Comparison between BALB/cJ and BALB/cByJ mice in tests of social behavior and resident-intruder aggression. Aggress Behav 15:273–280
Holick KA, Lee DC, Hen R, Dulawa SC (2008) Behavioral effects of chronic fluoxetine in BALB/cJ mice do not require adult hippocampal neurogenesis or the serotonin 1A receptor. Neuropsychopharmacology 33:406–417. doi:10.1038/sj.npp.1301399
JAX Notes (1990) A brief history of the two substrains of BALB/c, BALB/cJ, and BALB/cByJ. Available from Animal Resources. 443. http://jaxmice.jax.org/jaxnotes/archive/443a.html
Jiao J, Nitzke A, Doukas D et al (2011) Antidepressant response to chronic citalopram treatment in eight inbred mouse strains. Psychopharmacology 213:509–520
Keane TM, Goodstadt L, Danecek P et al (2011) Mouse genomic variation and its effect on phenotypes and gene regulation. Nature 477:289–294. doi:10.1038/nature10413
Kessler S, Elliott GR, Orenberg EK, Barchas JD (1977) A genetic analysis of aggressive behavior in two strains of mice. Behav Genet 7:313–321
Koboldt DC, Steinberg KM, Larson DE et al (2013) The next-generation sequencing revolution and its impact on genomics. Cell 155:27–38. doi:10.1016/j.cell.2013.09.006
Kumar V, Kim K, Joseph C et al (2013) C57BL/6 N mutation in Cytoplasmic FMRP interacting protein 2 regulates cocaine response. Science 342:1508–1512. doi:10.1126/science.1245503
Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25:1754–1760. doi:10.1093/bioinformatics/btp324
Li H, Handsaker B, Wysoker A et al (2009) The sequence alignment/map format and SAM tools. Bioinformatics 25:2078–2079. doi:10.1093/bioinformatics/btp352
Li H, Dow HC, Tan J et al (2012) Sociability and brain development in BALB/cJ and C57BL/6 J mice. Behav Brain Res 228:299–310. doi:10.1016/j.bbr.2011.12.001
Mehta M, Schmauss C (2011) Strain-specific cognitive deficits in adult mice exposed to early life stress. Behav Neurosci 125:29–36. doi:10.1037/a0021952
Miczek KA, O’Donnell JM (1978) Intruder-evoked aggression in isolated and nonisolated mice: effects of psychomotor stimulants and L-dopa. Psychopharmacology 57:47–55. doi:10.1007/BF00426957
Norcross M, Mathur P, Enoch AJ et al (2008) Effects of adolescent fluoxetine treatment on fear-, anxiety- or stress-related behaviors in C57BL/6 J or BALB/cJ mice. Psychopharmacology 200:413–424. doi:10.1007/s00213-008-1297-2
Palmer AA, Dulawa SC (2010) Murine Warriors or Worriers: the saga of comt1, B2 SINE elements, and the future of translational genetics. Front Neurosci 4:177. doi:10.3389/fnins.2010.00177
Potter M (1985) History of the BALB/c family. Curr Top Microbiol Immunol 122:1–5
Poulter MO, Du L, Zhurov V et al (2010) Plasticity of the GABA(A) receptor subunit cassette in response to stressors in reactive versus resilient mice. Neuroscience 165:1039–1051. doi:10.1016/j.neuroscience.2009.11.028
Sankoorikal GMV, Kaercher KA, Boon CJ et al (2006) A mouse model system for genetic analysis of sociability: C57BL/6 J versus BALB/cJ inbred mouse strains. Biol Psychiatry 59:415–423. doi:10.1016/j.biopsych.2005.07.026
Savignac HM, Dinan TG, Cryan JF (2011) Resistance to early-life stress in mice: effects of genetic background and stress duration. Front Behav Neurosci 5:13. doi:10.3389/fnbeh.2011.00013
Takahashi JS, Shimomura K, Kumar V (2008) Searching for genes underlying behavior: lessons from circadian rhythms. Science 322:909–912. doi:10.1126/science.1158822
Tarantino LM, Sullivan PF, Meltzer-Brody S (2011) Using animal models to disentangle the role of genetic, epigenetic, and environmental influences on behavioral outcomes associated with maternal anxiety and depression. Front Psychiatry 2:44. doi:10.3389/fpsyt.2011.00044
Velez Lady, Sokoloff G, Miczek KA et al (2010) Differences in aggressive behavior and DNA copy number variants between BALB/cJ and BALB/cByJ substrains. Behav Genet 40:201–210. doi:10.1007/s10519-009-9325-5
Vijayvargiya N, Fairless AH, Abel T et al (2013) Development of home cage social behaviors in BALB/cJ vs. C57BL/6 J mice. Behav Brain Res 237:338–347. doi:10.1016/j.bbr.2012.08.051
Wahlsten D (1974) Heritable aspects of anomalous myelinated fibre tracts in the forebrain of the laboratory mouse. Brain Res 68:1–18. doi:10.1016/0006-8993(74)90530-7
Wahlsten D (1989a) Genetic and developmental defects of the mouse corpus callosum. Experientia 45:828–838. doi:10.1007/BF01954057
Wahlsten D (1989b) Deficiency of the corpus callosum: incomplete penetrance and substrain differentiation in BALB/c mice. J Neurogenet 5:61–76. doi:10.3109/01677068909167265
Wahlsten D, Colbourne F, Pleus R (2003) A robust, efficient and flexible method for staining myelinated axons in blocks of brain tissue. J Neurosci Methods 123:207–214. doi:10.1016/S0165-0270(02)00359-X
Wang L, Jiao J, Dulawa SC (2011) Infant maternal separation impairs adult cognitive performance in BALB/cJ mice. Psychopharmacology 216:207–218. doi:10.1007/s00213-011-2209-4
Williams R, Lim JE, Harr B et al (2009) A common and unstable copy number variant is associated with differences in Glo1 expression and anxiety-like behavior. PLoS ONE 4:e4649. doi:10.1371/journal.pone.0004649
Yalcin B, Wong K, Agam A et al (2011) Sequence-based characterization of structural variation in the mouse genome. Nature 477:326–329. doi:10.1038/nature10432
Acknowledgments
This study was supported by R01MH079103 to AAP, R25GM066522 to CBC, and a Samsung Scholarship to CJ.
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Sittig, L.J., Jeong, C., Tixier, E. et al. Phenotypic instability between the near isogenic substrains BALB/cJ and BALB/cByJ. Mamm Genome 25, 564–572 (2014). https://doi.org/10.1007/s00335-014-9531-1
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DOI: https://doi.org/10.1007/s00335-014-9531-1