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| Published in final edited form as: J Int Neuropsychol Soc. 2009 September ; 15(5): 671–683. doi:10.1017/S1355617709990233. Molecular neurodevelopment: An in vivo31P-1H MRSI study Gerald Goldstein1, Kanagasabai Panchalingam2, Richard J. McClure2, Jeffrey A. Stanley6, Vince D. Calhoun7,8, Godfrey D. Pearlson9, and Jay W. Pettegrew2,3,4,5 1VA Pittsburgh Healthcare System, Pittsburgh, PA 2Department of Psychiatry, University of Pittsburgh School of Medicine, University of Pittsburgh, Pittsburgh, PA 3Department of Neurology, University of Pittsburgh School of Medicine, University of Pittsburgh, Pittsburgh, PA 4Department of Behavioral and Community Health Sciences, University of Pittsburgh School of Medicine, University of Pittsburgh, Pittsburgh, PA 5Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA 6Psychiatry and Behavioral Neurosciences, Wayne State University School of Medicine, Detroit, MI 7The Mind Research Network, Albuquerque, NM 8Department of Electrical and Computer Engineering, University of New Mexico, Albuquerque, NM 9Department of Psychiatry, Yale University, Hartford, CT Abstract Synaptic development and elimination are normal neurodevelopmental processes which if altered could contribute to various neuropsychiatric disorders. 31P-1H magnetic resonance spectroscopic imaging and structural MRI exams were conducted on 106 healthy children ages 6–18 years in order to identify neuromolecular indices of synaptic development and elimination. Over the age range studied, age-related changes in high-energy phosphate (phosphocreatine), membrane phospholipid metabolism (precursors and breakdown products), and gray matter were found. These neuromolecular and structural indices of synaptic development and elimination are associated with development of several cognitive domains and changes in gray matter volume. Monitoring of these molecular markers is essential for devising treatment strategies for neurodevelopmental disorders. Keywords MRS; Neuroimaging; Metabolism; Cognition; Multiple Regression INTRODUCTION The four major stages that characterize human brain development are: 1) neuronal proliferation; 2) migration of neurons to specific sites throughout the CNS; 3) organization of the neuronal circuitry; and 4) myelination of the neuronal circuitry (Volpe, 1995). |
The third stage of human brain development, organization of the neural circuitry, is most active from the sixth month of gestation to young adulthood. The major events associated with neuronal circuitry organization include: 1) proper alignment, orientation, and layering of cortical neurons; 2) dendritic and axonal differentiation; 3) synaptic development; 4) synaptic elimination (cell death and/or selective elimination of neuronal processes); and 5) glial proliferation and differentiation. These processes overlap with the timing of normal development of cognitive function and the onset of neurodevelopmental and psychiatric disorders such as attention deficit disorder, autism and schizophrenia. Normal synaptic elimination occurs during early adolescence in non-human primates (Rakic et al., 1986; Bourgeois & Rakic, 1993) and humans (Huttenlocher, 1979; Huttenlocher et al., 1982; Huttenlocher, 1990; Huttenlocher & Dabholkar, 1997). Synaptic elimination in non-human primates is generally observed to occur synchronously in all regions (i.e., homochronous) (Rakic et al., 1986) but is heterochronous in humans (Huttenlocher et al., 1997). Normal synaptic elimination is predominantly of presumptive excitatory asymmetric junctions on dendritic spines (Smiley & Goldman-Rakic, 1993) which probably utilize amino acids, such as L-glutamate, as the neurotransmitter (Storm-Mathisen & Otterson, 1990). Perinatal insults, intrauterine disturbances, and perhaps environmental influences in childhood and adolescence can potentially result in disordered neuronal circuitry (Birch & Gussow, 1970). This study focuses on molecular and structural indices related to synaptic development and elimination.
Conclusions
These conclusions are based on cross-sectional analysis and there would be clear advantages to testing our subjects longitudinally. We have done so, and will report the findings in a future publication. We would note though that it is not likely that the cross-sectional data were markedly influenced by cohort effects since all subjects were recruited from the same community over a period of only four years. There is the possibility of gender differences which we could not evaluate here because of small sample sizes. A final consideration is that the study was limited to an axial slice of brain measures of PCr, sPME, sPDE, and NAA and different results may be found at different regional locations. Since we had the capability of obtaining MRS measures from various brain regions, we will present the findings in a future report.
Acknowledgments
This work was supported in part by an NIHCD/NIH HD-39799 grant (JWP). We thank Terry Bradbury for conducting neuropsychological testing. Indebtedness is also expressed to the Medical Research Service and the VISNIV Mental Illness, Research, Education and Clinical Center (MIRECC), Department of Veterans Affairs for support of this work. We thank Harriet Marshman, Deborah Wetzler and Dennis McKeag for help in conducting the study.
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Figure 1.
(A) 31P MRSI voxel grid shifts (outlined in yellow) superimposed on the middle MRI axial slice (bottom right) for: a) prefrontal cortex; b) basil ganglia; c) superior temporal cortex; d) inferior parietal cortex; e) centrum semiovale; and f) occipital regions. Voxel size is 45×45×30 mm3. (B) Segmentation images of: a) gray matter; b) white matter; and c) CSF and extracortical matter where the intensity is proportional to the tissue type of that image. The matrix size of the images is 256 in the sagittal direction by 124 in the coronal direction, reflecting the 124 slices that were acquired for the 3-dimensional SPGR sequence. Right and left 31P prefrontal voxels (yellow boxes) are superimposed on the images.
Figure 2.
Quantification of a typical in vivo 31P magnetic resonance spectroscopic imaging spectrum with 5 Hz line broadening from a single voxel (30×30×30 mm3 ) of a study subject. The acquired spectrum is modeled in the time domain with Gaussian-damped sinusoids and by omitting both the first 0.75 ms and first 2.75 ms of the free induction decay using the MarquardtLevenberg algorithm. Both the short (0.75 ms) and long (2.75 ms) delay time (DT) models are shown superimposed on the acquired 31P spectra and the modeled resonances are identified on the right. The difference between the two time domain fits results in the bottom trace containing the intermediate correlation time components.
Figure 3.
An example of quantifying a short TE 1H MRSI spectrum of a control subject using the proposed acquisition protocol and LC Model fitting. The acquired spectrum with no line broadening is superimposed on the modeled and baseline spine function and the residual is below. The quantified macromolecule signal is indicated in a separate trace.
Figure 4.
(A) Scatterplots of PCr, sPME/sPDE, NAA, gray matter volume versus age fitted with a LOESS curve with 95% confidence intervals. (B) Scatterplots of composite scores for cognitive domains (Language, Memory, Visual Spatial, Executive Function) versus age fitted with a LOESS curve with 95% confidence intervals.
Figure 5.
Z-score plots of PCr, sPME/sPDE, NAA, and gray matter volume versus age.
Figure 6.
Z-score plots of PCr, gray matter volume, cognitive domain composite scores (Language, Memory, Visual Spatial, Executive Function).
Table 1
Neuropsychological test variables used within each cognitive domain.
DOMAIN TEST VARIABLES
Language Abbreviated WISC for children Total Verbal raw scores
Wechsler Individual Scale of Intelligence (WASI) for Total Verbal raw scores Adults
Wechsler Individual Achievement Test (WIAT) Reading, Spelling raw scores
Clinical Evaluation of Language Fundamentals (CELF) Concepts and Directions raw scores
Peabody Picture Vocabulary Tests Raw scores
| Executive Function | Wechsler Similarities and Matrix Reasoning Subtests Wisconsin Card Sorting Test | Raw scores Perseverative Errors |
| Visual Spatial | Wechsler Block Design | Raw score |
| Visual Motor Integration Test | Raw score | |
| Test of Visual Perception | Spatial Relations Subtest- raw score | |
| Memory | Wide Range Assessment of Memory and Learning (WRAML) | Picture Memory, Design Memory, Verbal Learning, Story Memory and Number/Letter Subtests |