Intrauterine hyperexposure to dexamethasone of the common marmoset monkey revealed normal cerebral metabolite concentrations in adulthood as assessed by quantitative proton magnetic resonance spectroscopy in vivo.

BACKGROUND: Animal models of human brain disorders often have to rely on non-human primates because of their immunological, physiological, and cognitive similarities to humans. METHODS: Localized proton magnetic resonance spectroscopy was performed to assess cerebral metabolite profiles of male common marmoset monkeys in vivo and to determine putative alterations of adult brain metabolism in response to intrauterine hyperexposure to the synthetic glucocorticoid hormone dexamethasone. RESULTS: Excellent spectral quality allowed for absolute quantification of the concentrations of major metabolites in predominantly white matter, gray matter, and thalamus. Marmoset monkeys intrauterinely hyperexposed to dexamethasone revealed normal neurochemical profiles at adulthood. CONCLUSIONS: Prenatally applied dexamethasone does not lead to persistent metabolic alterations affecting adult brain integrity.

Compatibility of glass-guided recording microelectrodes in the brain stem of squirrel monkeys with high-resolution 3D MRI.

Knowledge of the precise position of recording microelectrodes within the brain of a non-human primate is essential for a reliable exploration of very small anatomic structures. This work demonstrates the compatibility of a newly developed glass-guided microelectrode design and microfeed equipment with high-resolution 3D magnetic resonance imaging (MRI). T1- and T2-weighted images allow for the non-invasive visualization of chronically implanted microelectrodes within the brain stem of squirrel monkeys in vivo. Neural extracellular multi-unit recordings proved the functionality of the microelectrode before and after the use of 3D MRI suggesting the preservation of normal brain tissue at the tip of the electrode. Because histology confirmed the absence of lesions attributable to MRI, the approach offers an interactive monitoring during the course of neuroethological experiments. Consequently, MRI may become an in vivo alternative to common histological post mortem verifications of electrode tracks and hence may avoid the early sacrificing of primates after only a small number of experiments.

In vivo diffusion tensor mapping of the brain of squirrel monkey, rat, and mouse using single-shot STEAM MRI.

The purpose was to assess the potential of half Fourier diffusion-weighted single-shot STEAM MRI for diffusion tensor mapping of animal brain in vivo. A STEAM sequence with image acquisition times of about 500 ms was implemented at 2.35 T using six gradient orientations and b values of 200, 700, and 1200 s mm(-2). The use of half Fourier phase-encoding increased the signal-to-noise ratio by 45% relative to full Fourier acquisitions. Moreover, STEAM-derived maps of the relative anisotropy and main diffusion direction were completely free of susceptibility-induced signal losses and geometric distortions. Within measuring times of 3 h, the achieved resolution varied from 600x700x1000 microm3 for squirrel monkeys to 140x280x720 microm3 for mice. While in monkeys the accessible white matter fiber connections were comparable to those reported for humans, detectable fiber structures in mice focused on the corpus callosum, anterior commissure, and hippocampal fimbria. In conclusion diffusion-weighted single-shot STEAM MRI allows for in vivo diffusion tensor mapping of the brain of squirrel monkeys, rats, and mice without motion artifacts and susceptibility distortions.

Magnetization transfer MRI of mouse brain reveals areas of high neural density.

Extending applications of magnetization transfer contrast (MTC) in magnetic resonance imaging (MRI) of the human central nervous system, this work quantitatively describes MTC of the murine brain. As a novel finding, complementing T(1)- and T(2)-weighted MRI, MTC allows for the distinction of densely packed gray matter from normal gray and white matter. Examples include the Purkinje cell layer and the granular cell layer in the mouse cerebellum as well as the delineation of the CA3 subfield of the hippocampus relative to surrounding hippocampal gray matter and white matter tracts such as the hippocampal fimbria. Using a kainate lesion model, the CA3 hyperintensities in MTC and T(1)-weighted MRI are assigned to the densely packed somata of pyramidal cells.

Substance P receptor antagonist and clomipramine prevent stress-induced alterations in cerebral metabolites, cytogenesis in the dentate gyrus and hippocampal volume.

The neuropeptide substance P and its receptor, the neurokinin 1 receptor (NK(1)R) have been proposed as possible targets for new antidepressant therapies. The present study investigated the effect of the NK(1)R antagonist L-760,735 and the tricyclic antidepressant clomipramine in the chronic psychosocial stress paradigm of adult male tree shrews. Animals were subjected to a 7-day period of psychosocial stress before the onset of daily oral administration of L-760,735 (10 mg kg(-1) day(-1)) or clomipramine (50 mg kg(-1) day(-1)). The psychosocial stress continued throughout the treatment period of 28 days. Brain metabolite concentrations were determined in vivo by proton magnetic resonance spectroscopy. Cell proliferation in the dentate gyrus and hippocampal volume were measured post mortem. Stress significantly decreased in vivo concentrations of N-acetyl-aspartate (-14%), creatine and phosphocreatine (-15%) and choline-containing compounds (-15%). The proliferation rate of the granule precursor cells in the dentate gyrus was reduced (-45%), and hippocampal volume was decreased (-14%). The stress-induced changes of brain metabolites, hippocampal volume and dentate cytogenesis rate were prevented by concomitant drug administration. Elevated myo-inositol concentrations after both treatments hint to an astrocytic enhancement. These results suggest that-despite a different pharmacological profile-the NK(1)R antagonist L-760,735, a member of a novel class of antidepressant drugs, has comparable neurobiological efficacy to tricyclic antidepressants such as clomipramine.

High-resolution 3D MRI of mouse brain reveals small cerebral structures in vivo.

This work demonstrates technical approaches to high-quality magnetic resonance imaging (MRI) of small structures of the mouse brain in vivo. It turns out that excellent soft-tissue contrast requires the reduction of partial volume effects by using 3D MRI at high (isotropic) resolution with linear voxel dimensions of about 100-150 microm. The long T(2)* relaxation times at relatively low magnetic fields (2.35 T) offer the benefit of a small receiver bandwidth (increased signal-to-noise) at a moderate echo time which together with the small voxel size avoids visual susceptibility artifacts. For measuring times of 1-1.5 h both T(1)-weighted (FLASH) and T(2)-weighted (Fast Spin-Echo) 3D MRI acquisitions exhibit detailed anatomical insights in accordance with histological sections from a mouse brain atlas. Preliminary applications address the identification of neuroanatomical variations in different mouse strains and the use of Mn(2+) as a T(1) contrast agent for neuroaxonal tracing of fiber tracts within the mouse visual pathway.

Synaptic plasticity and tianeptine: structural regulation.

Stress-induced structural and cellular alterations in the hippocampus can contribute to the pathophysiology of depression. The reversal of these alterations may be a mechanism by which antidepressants achieve their therapeutic effect. The aim of the present study was therefore to investigate the effect of tianeptine on stress-induced structural changes and alterations in cerebral metabolites. To this end, psychosocially stressed male tree shrews were treated with tianeptine. A combination of in vivo and postmortem methods was used to evaluate the antidepressant treatment on the preservation of neuronal plasticity. It was found that all stress-induced effects were prevented by the administration of tianeptine. It is concluded that these findings provide experimental evidence for recent theories that impairment of neuronal viability and neuroplasticity might be important causal factors in mood disorders, suggesting tianeptine as a potential stimulator of neural resilience.

Gender-specific alterations of cerebral metabolites with aging and cortisol treatment.

Excess availability of the adrenocortical glucocorticoid hormone cortisol has been correlated with structural brain changes and a decline of cognitive functions during aging. Pertinent studies need to consider gender as a potential confound because of sexual dimorphism in the regulation of hypothalamus-pituitary-adrenal axis activity. In vivo localized proton magnetic resonance spectroscopy of male and female tree shrews revealed similar concentrations of cerebral metabolites in young adult animals but gender-specific alterations with aging as well as in response to cortisol treatment. In comparison with adult tree shrews, aged males had reduced concentrations of N-acetylaspartate (-33%; P<0.01) and total creatine (-34%; P< 0.01). These findings are in line with the occurrence of neuronal loss. In contrast, aged females exhibited increased concentrations of choline-containing compounds (+27%; P<0.05) which--together with a tendency for increased creatine (+24%) and myo-inositol (+14%)--is indicative of glial proliferation. After chronic administration of cortisol (4 mg/day for 28 days), male but not female tree shrews showed a specific reduction of the choline-containing compounds (-29%; P< 0.05). The observed sex differences with age are likely to result from differences in the regulation of stress-related hormones which is further supported by the gender-specific responses to cortisol.

Stress-induced changes in cerebral metabolites, hippocampal volume, and cell proliferation are prevented by antidepressant treatment with tianeptine.

Stress-induced structural remodeling in the adult hippocampus, involving debranching and shortening of dendrites and suppression of neurogenesis, provides a cellular basis for understanding the impairment of neural plasticity in the human hippocampus in depressive illness. Accordingly, reversal of structural remodeling may be a desirable goal for antidepressant therapy. The present study investigated the effect of tianeptine, a modified tricyclic antidepressant, in the chronic psychosocial stress model of adult male tree shrews (Tupaia belangeri), a model with high validity for research on the pathophysiology of major depression. Animals were subjected to a 7-day period of psychosocial stress to elicit stress-induced endocrine and central nervous alterations before the onset of daily oral administration of tianeptine (50 mg/kg). The psychosocial stress continued throughout the treatment period of 28 days. Brain metabolite concentrations were determined in vivo by proton magnetic resonance spectroscopy, cell proliferation in the dentate gyrus was quantified by using BrdUrd immunohistochemistry, and hippocampal volume was measured post mortem. Chronic psychosocial stress significantly decreased in vivo concentrations of N-acetyl-aspartate (-13%), creatine and phosphocreatine (-15%), and choline-containing compounds (-13%). The proliferation rate of the granule precursor cells in the dentate gyrus was reduced (-33%). These stress effects were prevented by the simultaneous administration of tianeptine yielding normal values. In stressed animals treated with tianeptine, hippocampal volume increased above the small decrease produced by stress alone. These findings provide a cellular and neurochemical basis for evaluating antidepressant treatments with regard to possible reversal of structural changes in brain that have been reported in depressive disorders.

Mapping of retinal projections in the living rat using high-resolution 3D gradient-echo MRI with Mn2+-induced contrast.

This study describes the neuroaxonal tracing of the visual pathway in the living rat using high-resolution T1-weighted 3D gradient-echo MRI (195 x 195 x 125 microm3) at 8, 24, 48, and 72 h after intraocular Mn2+ injection (0.1 microl of 1 M aqueous MnCl2). Best results were obtained at 24 h postinjection, revealing a continuous pattern of anterograde labeling from the retina, optic nerve, and chiasm to the contralateral optic tract, the dorsal and ventral lateral geniculate nucleus, the superior colliculus and its brachium, the olivary pretectal nucleus, the nucleus of the optic tract, and the suprachiasmatic nucleus. These results underline the feasibility of repeated MRI tract tracing in living animals after a single injection of Mn2+. The approach is expected to advance studies of neuroaxonal function in behaving animals with special emphasis on applications in developmental neurobiology.

Psychosocial stress, glucocorticoids, and structural alterations in the tree shrew hippocampus.

Animal models for chronic stress represent an indispensable preclinical approach to human pathology since clinical data point to a major role of psychological stress experiences, acute and/or chronic, to the development of behavioral and physiological disturbances. Chronic emotional arousal is a consequence of various types of social interactions, and one major neurohumoral accompaniment is the activation of the classic stress circuit, the limbic--hypothalamic--pituitary--adrenocortical (LHPA) axis. The adrenocortical glucocorticoid hormones cortisol and corticosterone are principal effectors within this circuit since they affect neurotransmission and neuroendocrine control, thus having profound effects on mood and behavior. Using the experimental paradigm of chronic psychosocial stress in tree shrews, we investigated the impact of aversive chronic social encounters on hippocampal structure and function. In chronically stressed animals, we observed dendritic atrophy of hippocampal pyramidal neurons and an impairment of neurogenesis in the dentate gyrus. However, a stress-induced loss of hippocampal neurons was not observed in this animal model. This review summarizes our recent results on structural changes occurring during chronic stress in neurons of the hippocampus and their potential influence on learning and memory. We discuss whether these changes are reversible and to what extent glucocorticoids might be responsible for the stress-induced effects.

Effect of chronic psychosocial stress and long-term cortisol treatment on hippocampus-mediated memory and hippocampal volume: a pilot-study in tree shrews.

We investigated the impact of chronic psychosocial stress and long-term cortisol treatment on hippocampus-mediated memory processes and hippocampal volume in male tree shrews. By combining cognitive tests on a hole board, magnetic resonance imaging (MRI), and saliva cortisol analysis, we were able to follow in individual animals the stress- and cortisol-induced temporal effects on HPA axis activity and the hippocampus and its executive functions during 15 weeks. Four weeks of either cortisol treatment or psychosocial stress affected hippocampus-mediated memory. Cortisol-induced impairments were observed only at the end of the treatment phase while in stressed animals a negative effect on hippocampus-mediated memory was monitored after 7 weeks of recovery. A trend towards a reduction of the hippocampal volume was found in both experimental groups. The present preclinical study demonstrated that major life events, such as psychosocial stress, or chronic cortisol treatment leave traces in hippocampus-dependent memory. This finding requires systematic analysis to understand how brain areas critical for information processing such as the hippocampal formation are affected by stress and stress hormones.

Brain water diffusion in normal and creatine-supplemented rats during transient global ischemia.

Brain water diffusion in response to transient global ischemia (12 min), reperfusion (60 min), and cardiac arrest was monitored by localized proton magnetic resonance spectroscopy. The trace of the apparent diffusion coefficient tensor (ADC(Av)) was determined at high temporal resolution (10 sec) to assess the putative neuroprotective potential of oral creatine (Cr) in rats that received 2.2 g Cr-monohydrate per kg body weight per day for 10 days (n = 8) relative to controls (n = 9). Cr-fed rats revealed a statistically significant increase of the cerebral concentration ratio of Cr to choline-containing compounds (20%). The decrease of the ADC(Av) value during acute ischemia showed a three-phasic behavior in line with energy depletion, cytotoxic edema, and brain cooling. In Cr-fed rats, slightly less severe and mildly delayed diffusion changes during ischemia and similar beneficial trends during early reperfusion did not reach statistical significance. Magn Reson Med 42:798-802, 1999.

Proton MRS of oral creatine supplementation in rats. Cerebral metabolite concentrations and ischemic challenge.

Proton magnetic resonance spectroscopy (MRS) was employed to determine the concentrations of N-acetylaspartate (NAA), total creatine (tCr), choline-containing compounds (Cho), myo-inositol (Ins), glucose (Glc), and lactate (Lac) in rat brain before and after 10 days of oral supplementation of 2.6 g Cr-monohydrate per kg body weight per day. Measurements were performed both in vitro (n = 16) and in vivo (n = 6). The neuroprotective potential of oral Cr was assessed by dynamically monitoring brain Glc and Lac in response to transient global ischemia (12 min). In comparison to controls the in vitro concentrations of Cr (13.1 +/- 9.3%) and Ins (12.7 +/- 14. 0%) were significantly increased in Cr-fed rats. Under in vivo conditions, the data revealed trends for elevated tCr (4.7%) and Ins (10.6%) which were enhanced in the concentration ratios of tCr:Cho (10.2%) and Ins:Cho (17.8%). Together with an increased Glc level (27.3%), the observation of a statistically significant decrease of brain Lac (-38.5 +/- 19.3%) in Cr-fed rats may reflect a shift of the energy metabolism from non-oxidative toward oxidative glycolysis. One hour after global ischemia most of the metabolic differences between Cr-fed rats and controls were retained. The increased Glc level (44.4 +/- 33.3%) reached statistical significance, but the accumulation of Lac and its time course during ischemia and early reperfusion showed no differences between Cr-fed rats and controls.

Volumetric MRI measurements of the tree shrew hippocampus.

Protocols suitable for repeated magnetic resonance imaging (MRI) studies of the tree shrew's brain were established. This included the development of (i) a technique for prolonged inhalation anesthesia by endotracheal intubation; (ii) a reproducible fixation of the animal's head in a stereotaxic frame and finally (iii) the set-up of the hardware (rf coil) and software (MRI sequences) of the MRI system. The endotracheal intubation as well as the repeated and prolonged anesthesia showed no complications. The in vivo measurements of the tree shrew's hippocampal formation revealed a high reproducibility. Right and left hippocampal volume was determined as 85.2 mm3 +/- 8% and 87.4 mm3 +/- 10%, respectively. The utility of MRI in delineating alterations in brain anatomy was demonstrated in three animals receiving cortisol via the drinking water (5 mg/animal/day). After a 4-week treatment, in two of the three tree shrews a reduction in hippocampal volume was observed. Thus, the MRI protocols used here allow for repeated and non-invasive measurements of changes in hippocampal anatomy within the same animal and to monitor the temporal dynamics of structural alterations within this brain structure.

Proton T2 relaxation of cerebral metabolites during transient global ischemia in rat brain.

Putative changes of metabolite T2 relaxation times were investigated before and after a 20-min period of global ischemia in rat brain in vivo (n = 10) using localized proton MRS at different echo times (2.35 T). Neither absolute T2 relaxation times (TE = 20-270 ms) nor time courses of T2-weighted metabolite signals (TE = 135 ms) revealed statistically significant changes during the occlusion or early reperfusion relative to pre-ischemic baseline. These findings are in line with reports of relaxation changes at much later stages and further demonstrate that altered T2 relaxation is not a confounding factor in diffusion-weighted long-TE proton MRS during early ischemic events.

Metabolic alterations in brain autopsies: proton NMR identification of free glycerol.

Metabolic alterations in bovine brain homogenate were examined as a function of post mortem interval (PMI) using high-resolution proton NMR spectroscopy. In particular, while lactate, glutamate, glutamine, creatine and inositols as well as the total concentration of trimethyl-ammonium compounds remained constant, prominent changes due to the hydrolysis of N-acetylaspartate to acetate and aspartate as well as the decomposition of glycerophosphocholine into free choline and glycerol correlated linearly with the duration of PMI (3-195 h). The spectroscopic identification of the latter process was confirmed by proton NMR studies of model solutions as well as of extracts of mammalian brain showing high levels of free glycerol. Since the methylene resonances of glycerol overlap with the proton resonances of myo-inositol, care should be taken in the interpretation of both in vitro and in vivo brain spectra.

Dialysis and transplantation affect cerebral abnormalities of end-stage renal disease.

Localized short echo time proton magnetic resonance spectroscopy was performed to determine whether chronic and end-stage renal failure, hemodialysis, continuous ambulatory peritoneal dialysis, or renal transplantation result in alterations of cerebral water and metabolites in humans. Hemodialysis patients show an increased cerebral concentration of myo-inositol (+ 14%; P < .05). Increased metabolite ratios are found for myo-inositol/creatine (+14%; P <.01) and choline containing compounds choline/creatine (+10%; P < .01) and are more marked in gray than in white matter. N-acetylaspartate and total creatine concentrations are unaffected. Compared to hemodialysis, continuous ambulatory peritoneal dialysis patients show a larger increase in choline and less elevated myo-inositol. Acutely, hemodialysis significantly decreases the cerebrospinal fluid content of the examined brain regions, but metabolite changes are small compared to the persistent alterations in patients receiving hemodialysis or continuous ambulatory peritoneal dialysis. Undialyzed patients with chronic renal failure do not differ from patients on hemodialysis, but cerebral metabolite changes are completely reversed by transplantation. Cerebral metabolic effects of end-stage renal disease differ from Alzheimer's disease, which is associated with markedly reduced N-acetylaspartate, increased myo-inositol, and normal choline concentrations. The small but significant cerebral metabolic disorders associated with renal failure and dialysis may be a consequence of osmotic dysregulation.

Probable Alzheimer disease: diagnosis with proton MR spectroscopy.

PURPOSE: To distinguish probable Alzheimer disease (AD) from other dementias (ODs) and normality in the elderly. MATERIALS AND METHODS: A double-blind trial of proton magnetic resonance (MR) spectroscopy was performed, principally in gray matter, in the occipital cortex of 114 patients with dementia (AD [n = 65], OD [n = 39], or frontal lobe dementia [FLD] [n = 10]), 98 patients without dementia, and 32 healthy control subjects. RESULTS: Reduced levels of N-acetylaspartate (NAA) (P < .0005) and increased levels of myo-inositol (MI) (P < .0005) characterize AD. Patients with OD had significantly reduced levels of NAA (P < .01) but normal levels of MI (P [vs AD] < .0005). When MI/NAA was used, AD was distinguished from normality with 83% sensitivity and 98% specificity. When MI/creatine was used, OD was distinguished from AD and FLD with a negative predictive rate of 80%, sensitivity of 82%, and specificity of 64%. CONCLUSION: Hydrogen-1 MR spectroscopy enables identification of mild to moderate AD with a specificity and sensitivity that suggest clinical utility.