Methodological and Stereological Considerations in Postmortem Psychiatric Brain Research

  • Ian Paul Everall
  • Paul J. Harrison
Part of the Neurobiological Foundation of Aberrant Behaviors book series (NFAB, volume 4)


There has been a renewal of interest in understanding the brain changes that may underlie schizophrenia. However, microscopic assessment of the brain is not a trivial task and there are many potential factors, which can affect experimental results. In this chapter we will review the major potential confounding variables to be considered and principles of quantifying microscopic indices such as cell number and size. In this regard the basic principles of stereology will be outlined as well as the basic ideas of quantifying cellular arrangement by the newly emerging tool of spatial pattern analysis. During these description examples from the published literature will be provided.


Major Depressive Disorder Superior Temporal Gyrus Postmortem Interval Complete Spatial Randomness Metachromatic Leukodystrophy 
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  1. Abusaad I, MacKay D, Zhao J, Stanford P, Collier DA, Everall IP. Stereological estimation of the total number of neurons in the murine hippocampus using the optical dissector. Journal of Comparative Neurology 1999; 408: 560–566.PubMedCrossRefGoogle Scholar
  2. Aherne WA, Dunnill MS. Morphometry. London: Edward Arnold. 1982Google Scholar
  3. Arnold SE, Gur RE, Shapiro RM, Fisher KR, Moberg PJ, Gibney MR, Gur RC, Blackwell P, Trojanowski JQ. Prospective clinicopathologic studies of schizophrenia: accrual and assessment of patients. American Journal of Psychiatry 1995; 152: 731–737.PubMedGoogle Scholar
  4. Barton AJL, Pearson RCA, Najlerahim A, Harrison PJ. Pre- and postmortem influences on brain RNA. Journal of Neurochemistry 1993; 61: 1–11.PubMedCrossRefGoogle Scholar
  5. Benes FM, Lange N. Two-dimensional versus three-dimensional cell counting a practical perspective. Trends in Neuroscience 2001; 24: 11–17.CrossRefGoogle Scholar
  6. Black JE, Kodish IM, Klintsova AY, Greenough WT, Uranova NA Quantitative study of human prefrontal pyramidal and interneurons: synaptic connectivity effects of schizophrenia and experience [abstract]. Schizophrenia Research 2001; 49 (suppl) 59.CrossRefGoogle Scholar
  7. Bonin G von. About quantitative studies in the cerebral cortex, Part I. Journal of Microscopy 1973; 99: 75–83.CrossRefGoogle Scholar
  8. Cairns NJ, Lantos PL. Brain tissue banks in psychiatric and neurological research Journal of Clinical Pathology 1996; 49: 870–873.Google Scholar
  9. Corsellis JAN Psychoses of obscure pathology. In: Greenfield’s Neuropathology, 3`d edition (eds Blackwood H, Corsellis JAN ). Edward Arnold, London, 1976: pp 903–915.Google Scholar
  10. Cotter D, Miskuel K, Al-Sarraj S, Wilkinson I, Paley M, Harrison MJ. & Everall, I. The investigation of postmortem brain volume using MRI: a comparison of planimetric and stereological approaches. J Neuroradiol 1999; 41: 493–496.CrossRefGoogle Scholar
  11. Cotter D, MacKay D, Beasley C, Kerwin R, Everall I. Reduced glial density and neuronal volume in major depressive disorder and schizophrenia in the anterior cingulate cortex. 106 Biennial Workshop on Schizophrenia, Schizophrenia Research 2000a; 41: 106.CrossRefGoogle Scholar
  12. Cotter D, Beasley CL, Kerwin R, Everall IP. The spatial pattern of parvalbumin immunoreactive neurons in the anterior cingulate cortex (BA 24b/c) in schizophrenia and bipolar disorder. 10th Biennial Workshop on Schizophrenia, Schizophrenia Research 2000b; 41: 107.Google Scholar
  13. Cruz Orive LM. On the estimation of particle number. Journal of Microscopy 1980; 120: 1527.Google Scholar
  14. Cullen TJ, Walker MA, Roberts H, Crow TJ, Harrison P, Esiri M. The mediodorsal nucleus of the thalamus in schizophrenia: a postmortem study. 106 Biennial Workshop on Schizophrenia, Schizophrenia Research 2000; 41: 5.CrossRefGoogle Scholar
  15. Davies J, Everall IP and Lantos PL. The contemporary AIDS database and brain bank lessons from the past. Journal of Neural Transmission 1993; 39 77–85.PubMedGoogle Scholar
  16. Davies J, Everall IP, Weich S, McLaughlin J, Scaravilli F and Lantos PL. HIV-associated brain pathology in the United Kingdom: an epidemiological study. AIDS 1997; 11 1145–1150.PubMedCrossRefGoogle Scholar
  17. Davies J, Everall, IP, Weich S, Glass J, Sharer LR, Cho ES, Bell JE, Mattenyi C, Gray F, Scaravilli F, Lantos PL. HIV-associated brain pathology: a comparative international study. Neuropathology and Applied Neurobiology 1998; 24 118–124.PubMedCrossRefGoogle Scholar
  18. Delesse MA. Procede mechanique pour determines la composition des roches (extrait). CR Academy of Science (Paris) 1847; 25: 544.Google Scholar
  19. Diggle Pi, Lange N, Benes FM. Analysis of variance for replicated spatial point patterns in clinical neuroanatomy. Journal of the American Statistical Association 1991; 86: 618–625.CrossRefGoogle Scholar
  20. Diggle PJ. Statistical analysis of spatial point patterns. Academic Press. New York. 1983 Eastwood SL, Harrison PJ Hippocampal synaptic pathology in schizophrenia, bipolar disorder and unipolar depression: a study of complexin mRNAs. Molecular Psychiatry 2000; 5 425–432Google Scholar
  21. Efron B. The jackknife, the bootstrap and other resampling plans. Society for Industrial and Applied Mathematics. CBMS-NSF Monograph 38. Philadelphia. 1983Google Scholar
  22. Gundersen HJ. The nucleator. Journal of Microscopy 1988; 151: 3–21.CrossRefGoogle Scholar
  23. Gundersen HJ, Osterby R. Optimizing sampling efficiency of stereological studies in biology: or ‘do more less well!’. Journal of Microscopy 1981; 121: 65–73.PubMedCrossRefGoogle Scholar
  24. Gundersen HJG, Bendtsen TF, Korbo L, Marcussen N, Moller A, Nielsen K, Nyengaard JR, Pakkenberg B, Sorensen FB, Vesterby A, West MJ. Some new simple efficient stereological methods and their use in pathological research and diagnosis. Acta Pathologica Microbiologica et Immunologica Scandinavica 1988a; 96: 379–394.CrossRefGoogle Scholar
  25. Gundersen HJG, Bagger P, Bendtsen TF, Evans SM, Korbo L, Marcussen N, Moller A, Nielsen K, Nyengaard JR, Pakkenberg B, Sorensen FB, Vesterby A, West MJ. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. Acta Pathologica Microbiologica et Immunologica Scandinavica 1988b; 96: 857–881.CrossRefGoogle Scholar
  26. Harris KM Structure, development, and plasticity of dendritic spines. Current Opinion in Neurobiology 2000; 9: 343–348Google Scholar
  27. Harrison PJ Advances in postmortem molecular neurochemistry and neuropathology: Examples from schizophrenia research. British Medical Bulletin 1996; 52: 527–538PubMedCrossRefGoogle Scholar
  28. Harrison PJ The neuropathological effects of antipsychotic drugs. Schizophrenia Research 1999; 40: 87–99PubMedCrossRefGoogle Scholar
  29. Harrison PJ, Kleinman JE Methdological issues. In: The Neuropathology of Schizophrenia: Progress and interpretation (eds PJ Harrison and GW Roberts ), Oxford University Press, Oxford, 2000: pp 339–350.Google Scholar
  30. Harrison PJ, Lewis DA The neuropathology of schizophrenia. In Schizophrenia, 2nd edition (eds Hirsch S, Weinberger DR). Blackwells, Oxford (in press), 2001Google Scholar
  31. Harrison PJ, Proctor AW, Barton AJL et al. Terminal coma affects messenger RNA detection in postmortem human temporal cortex. Molecular Brain Research 1991; 9: 161–164.PubMedCrossRefGoogle Scholar
  32. Harrison PJ, Barton AJL, Procter AW, Bowen DM, Pearson RCA The effects of Alzheimer’s disease, other dementias and premorten course upon amyloid ß precursor protein messenger RNAs in frontal cortex. Journal of Neurochemistry 1994; 62 635–644PubMedCrossRefGoogle Scholar
  33. Harrison Pi, Heath PR, Eastwood SL, Burnet PWJ, McDonald B, Pearson RCA The relative importance of premortem acidosis and postmortem interval for human brain gene expression studies: selective mRNA vulnerability and comparison with their encoded proteins. Neuroscience Letters 1995; 200: 151–154.PubMedCrossRefGoogle Scholar
  34. Haug H. History of Morphometry. Journal of Neuroscience Methods 1986; 18: 1–17.PubMedCrossRefGoogle Scholar
  35. Haug H, kuhl S, Mecke E, Sass N-L, Wasner K. The significance of morphometric procedures in the investigation of age changes in cytoarchitectonic structures of the human brain. Journal fur Hirnforschung 1984; 25: 353–374.PubMedGoogle Scholar
  36. Heckers S, Heinsen H, Geiger B, Beckmann H. Hippocampal neuron number in schizophrenia. A stereological study. Archives of General Psychiatry 1991; 48: 1002–1008.PubMedCrossRefGoogle Scholar
  37. Howard CV, Reed MG.Unbiased Stereology. Three Dimensional Measurement in Microscopy. Bios Scientific Publishers, Oxord, UK. 1998Google Scholar
  38. Hyman BT, Gomez-Isla T, Irizarry MC. Stereology: a practical primer for neuropathology. Journal of Neuropathology and Experimental Neurology 1998; 57: 305–310.PubMedCrossRefGoogle Scholar
  39. Neuropathology Group of the Medical Research Council Cognitive Function and Ageing Study (MRC CFAS) Pathological correlates of late-onset dementia in a multicentre, community-based population in England and Wales. Lancet 2001; 357, 169–175Google Scholar
  40. Johnston NL, Cerevnak J, Shore AD, Torrey EF, Yolken RH. Multivariate analysis of RNA levels from postmortem human brain as measured by three different methods of RT-PCR. Journal of Neuroscience Methods 1997; 77: 83–92.PubMedCrossRefGoogle Scholar
  41. Jones L, Mall N, Byne W. Localization of schizophrenia-associated thalamic volume loss. Society of Neuroscience Abstracts 1998; 24: 985Google Scholar
  42. Kingsbury AE, Foster OJ, Nisbet AP Cairns N, Bray L, Eve DJ, Lees AJ, Marsden CD.. Tissue pH as an indicator of mRNA preservation in human postmortem brain. Molecular Brain Research 1995; 28: 311–318.PubMedCrossRefGoogle Scholar
  43. Ongur D, Drevets WC, Price JL. Glial reduction in the subgenual prefrontal cortex in mood disorders. Proceedings of the Natlional Academy of Sciences USA. 1998; 95: 13290–13295.CrossRefGoogle Scholar
  44. Pakkenberg B. Pronounced reduction of total neuron number in mediodorsal thalamic nucleus and nucleus accumbens in schizophrenics. Archives of General Psychiatry. 1990; 47: 1023–1028.PubMedCrossRefGoogle Scholar
  45. Popken GJ, Bunney Jr WE, Potkin SG, Jones EG Subnucleus-specific loss of neurons in medial thalamus of schizophrenics. Proc Natl Acad Sci USA 2000; 97: 9276–9280PubMedCrossRefGoogle Scholar
  46. Rajkowska G, Miguel-Hidalgo JJ, Wei J, et al. Morphometric evidence for neuronal and glial prefrontal pathology in major depression. Biological Psychiatry 1999; 45: 1085–1098.PubMedCrossRefGoogle Scholar
  47. Ravid R, Swaab D The Netherlands brain bank — a clinico-pathological link in aging and dementia research. Journal of Neural Transmission 1993; 39: 143–153 supplementGoogle Scholar
  48. Ripley BD. The second-order analysis of stationary point processes. Journal of Applied Probability. 1976; 13: 255–266.CrossRefGoogle Scholar
  49. Roberts GW, Harrison PJ Gliosis and its implications for the disease process. In: The Neuropathology of Schizophrenia: Progress and interpretation (eds PJ Harrison and GW Roberts ), Oxford University Press, Oxford, 2000: pp 137–150.Google Scholar
  50. Selemon LD, Lidow MS, Goldman-Rakic PS Increased volume and glial density in primate prefrontal cortex associated with chronic antipsychotic drug exposure. Biological Psychiatry 1999: 46: 161–172.PubMedCrossRefGoogle Scholar
  51. Steno DC. The unbiased estimation of number and sizes of arbitrary particles using the disector. Journal of Microscopy 1984; 134: 127–136.CrossRefGoogle Scholar
  52. Stevens JR Enough of pooled averages: been there, done that. Biological Psychiatry 1997; 41: 633–635Google Scholar
  53. Stewart MG. Quantitative methods in neuroanatomy. John Wiley and Sons, Chichester. 1992 Torrey EF, Webster M, Knable M, Johnston N, Yolken RH. The Stanley Foundation Brain Collection and neuropathology consortium. Schizophrenia Research 2000; 44: 151–155.Google Scholar
  54. Upton GJG, Fingleton B. The Use of Mapped Plant Locations. In Spatial Data Analysis by Example Volume 1 ( Upton GJG, Fingleton B). John Wiley & Sons, Chichester, 1985: pp 70–93.Google Scholar
  55. Young KA, Manaye KF, Liang CL, Hicks PB, German DC Reduced number of mediodorsal and anterior thalamic neurons in schizophrenia. Biological Psychiatry 2000; 47: 944–953PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Ian Paul Everall
  • Paul J. Harrison

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