Department of Psychiatry, Education and Research Centre, Royal College of Surgeons in Ireland, Dublin, Ireland
Department of Neuroscience, Institute of Psychiatry
Section of the Neurobiology of Psychosis, Institute of Psychiatry
Department of Neuroscience, Institute of Psychiatry
Department of Biostatistics, Institute of Psychiatry, London, UK
Correspondence: Dr David R. Cotter, Beaumont Hospital, Dublin 9, Ireland. Tel: +353 1 809 3740; fax: +353 1 809 3741; e-mail: drcotter{at}rcsi.ie
Declaration of interest None. Funding detailed in Acknowledgements.
ABSTRACT
There is evidence that the superior temporal gyrus and Heschls gyrus within it are implicated in schizophrenia. We investigated neuronal and glial cell density and cortical thickness within Heschls gyrus, using the optical disector to estimate cell density within cortical layers 3 and 5 in tissue derived postmortem from people with diagnoses of major depressive disorder, schizophrenia and bipolar disorder, compared with normal controls (n=15 per group). No significant difference in neuronal or glial cell density or in cortical thickness was observed between the groups; our findings therefore provide no support for the presence of cellular pathology within Heschls gyrus in schizophrenia.
Heschls gyrus is located on the dorsal surface of the superior temporal gyrus and forms part of the primary auditory cortex. In schizophrenia the volume of the superior temporal gyrus and its posterior section, the planum temporale, are reduced (Barta et al, 1997) and both regions show activation during auditory hallucinations (Shergill et al, 2000). However, although Heschls gyrus forms the anterior border of the planum temporale and is functionally connected with it, few neuroimaging studies have focused on this gyrus and consistent changes have not been observed (Sumich et al, 2002). There has been no quantitative neuropathological study of this region in schizophrenia (Harrison, 1999). Therefore, we proposed to characterise neuronal and glial cell density and cortical thickness within Heschls gyrus in people with schizophrenia, major depressive disorder and bipolar disorder. We focused on layers 3 and 5 of this region because previous investigations have indicated that the pyramidal neurons of these layers (Pierri et al, 2001) might be particularly vulnerable in schizophrenia.
METHOD
Human brain specimens were obtained from the Stanley Foundation Brain Consortium. The specimens were derived from 60 individuals: 15 were normal controls, 15 had had a diagnosis of schizophrenia, 15 of bipolar disorder and 15 of major depressive disorder. Demographic, clinical and histological information relating to this sample is given by Torrey et al (2000).
Tissue was available from one hemisphere of each brain. Hemispheres were fixed in 10% phosphate buffered formalin, and the temporal lobes and superior temporal gyrus were dissected into a series of 1 cm-thick blocks. Each block was embedded in paraffin wax and tissue sections of 25 µm thickness were cut; one of these was randomly selected and stained with cresyl violet. This series of tissue sections, spanning the rostrocaudal extent of the superior temporal gyrus, was examined and sections containing Heschls gyrus (Brodmann area 41) were identified. Where more than one gyrus was present, only the most anterior was categorised as the Heschls gyrus. Four tissue sections that included Heschls gyrus were selected for examination from the specimens derived from each individual.
Neurons and glial cells within cortical layers 3 and 5 were identified by standard criteria and stereologically based quantitation was undertaken as described for a previous study of this brain series (Cotter et al, 2001). We used an optical disector of depth 10 µm with guard depths above and below of 3 µm. Cell density estimations were made with the aid of image analysis software (Kinetic Imaging version 5.0 for Windows, Histometrix, Liverpool, UK) according to the stereological optical disector method. The dimensions of the disector used were 50.5 µm x 37.5 µm in the x and y axes. We defined the cortical layers of interest by tracing the layer borders at x20 magnification, and a systematic random sampling strategy was employed within these. An average of 75 fields were counted per layer per case for neuronal and glial estimations, and an average of 169 neurons and 215 glia were counted in each layer of each case. Values for the coefficient of error of the density estimates were all less than 5%. Neuronal and glial cell densities are expressed as cells x 103 per mm3. The width of each cortical layer was assessed using an image analysis system (Image-Pro Plus 4.0 for Windows, Media Cybernetics, Silver Spring, Maryland, USA) and methods described by Chana et al (2003).
Statistical analysis
Neuronal and glial cell density in layers 3 and 5, and the thickness of
each of the six cortical layers, were analysed using the same statistical
methods applied in our previous study of this brain series
(Cotter et al, 2001).
A forward selection procedure with 10% inclusion threshold was used to
identify demographic and histological variables predicting outcome. Group
comparisons of cell densities and layer thickness were adjusted for potential
confounders and empirical predictors. For cortical thickness the
Bonferroni-adjusted significance level was 0.05/6=0.0083, and for cell density
the adjusted significance level was 0.05/2=0.025. For the analysis of cell
density we used Poisson modelling to compare densities between groups. The
random-effects Poisson models were fitted using the procedure GLMM in the
statistical package Genstat 5 (for Windows; VSN International Ltd), which
employs Schalls method to fit a generalised linear mixed model.
Accumulated analysis of deviance using the experimental method was used to
test for differences between the patient groups and the control group.
Standard multiple regression models were used to assess group differences in
layer thickness. All layer thickness analyses were carried out in the
Statistical Package for the Social Sciences, version 11 for Windows. Cortical
thickness data are summarised by their means, and densities by their
medians.
RESULTS
All modelling of layer thickness and cell density was adjusted for gender, postmortem interval and fixation time.
Cortical layer thickness
The forward selection procedure identified age as a predictor of total
cortical layer thickness (F(1, 55)=4.3, P=0.044),
with thickness estimated to decrease by 71.3 µm with every additional 10
years of age at death. Hence, layer-wise group comparisons were also adjusted
for age. No group difference in layer-wise cortical thickness was observed,
other than an increase in layer 4 in major depressive disorder
(P=0.047), but this effect could not be maintained after adjusting
for multiple layer-wise comparisons (data presented as a data supplement to
the online version of this paper and available from the authors on
request).
Cell density
Summaries of the neuronal and glial densities and the results of the
statistical comparisons are shown in Table
1. Tissue pH was found to be an empirical predictor of neuronal
density (Wald test 2(1)=12.1, P=0.0005) and glial
density (Wald test
2(1)=18.4, P<0.0001). Hence,
the layer-wise group comparisons were also adjusted for tissue pH. There was
no significant group difference in neuronal densities. We detected decreased
glial density in major depressive disorder relative to controls in layer 3
(P=0.038), but this finding did not remain significant after
adjusting for two layer-wise comparisons.
|
DISCUSSION
Our main findings are that cortical thickness and neuronal and glial cell densities in cortical layers 3 and 5 of Heschls gyrus are unaltered in schizophrenia and affective disorders. These are important negative findings for schizophrenia, as this region has been implicated in these disorders by neuroimaging investigations but has not yet been assessed using cytoarchitectural morphometric methods. We now demonstrate that there is no overt cellular abnormality within these cortical layers in this region in schizophrenia. Previous morphometric studies of bipolar disorder and major depression within prefrontal cortical regions have demonstrated deficits in glial cell densities and counts (Rajkowska et al, 1999; Cotter et al, 2001). Our negative cell counting data on Heschls gyrus in these disorders suggest that glial cell deficit in affective disorders might be localised to prefrontal cortical association regions, sparing primary sensory cortical regions. Taken together, these findings indicate that the cellular neuropathology of major psychiatric disorders demonstrates regional vulnerability. A current challenge is to determine the basis of such regional vulnerabilities.
ACKNOWLEDGMENTS
The study was funded by the Wellcome Trust, the Medical Research Council (UK) and the Theodore and Vada Stanley Foundation. Post-mortem brains were donated by the Stanley Foundation Brain Bank Consortium, courtesy of Drs Llewellyn B. Bigelow, Juraj Cervenak, Mary M. Herman, Thomas M. Hyde, Joel Kleinman, Jose D. Paltan, Robert M. Post, E. Fuller Torrey, Maree J. Webster and Robert Yolken.
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Received for publication May 8, 2003. Revision received December 8, 2003. Accepted for publication March 20, 2004.
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