Early neurological deterioration is associated with poor functional outcomes in stroke patients, but the underlying mechanisms remain unclear. This study aims to understand the progression of stroke-related brain damage using a rhesus monkey model with ischemic occlusion. Multiparameter MRI was used to monitor the progressive evolution of the brain lesion following stroke. Resting-state functional MRI, dynamic susceptibility contrast perfusion MRI, diffusion tensor imaging, and T1- and T2-weighted scans were acquired prior to surgery and at 4–6 h, 48 h, and 96 h following the stroke. The results revealed a sudden increase in infarction volume after the hyper-acute phase but before 48 h on diffusion-weighted imaging (DWI), with a slight extension by 96 h. Lower relative cerebral blood flow (CBF) and time to maximum (Tmax) prior to the stroke, along with a progressive decrease post-stroke, were observed when compared to other stroke monkeys in the same cohort. Functional connectivity (FC) in the ipsilesional secondary somatosensory cortex (S2) and primary motor cortex (M1) exhibited an immediate decline on Day 0 compared to baseline and followed by a slight increase on Day 2 and a further decrease on Day 4. These findings provide valuable insights into infarction progression, emphasizing the critical role of collateral circulation and its impact on early neurological deterioration during acute stroke.

Chronic psychosocial stress is associated with increased risk of psychiatric disorders. Magnetic resonance spectroscopy (MRS) in humans has been used to show that glutamate levels in medial prefrontal cortex (mPFC) following acute stress exposure adapt to recent chronic stress levels. Here, we sought to determine the presence of this glutamate stress response adaptation in rhesus macaques, whose societies are maintained by dominance relationships that are enforced by agonistic interactions and result in chronic stress phenotypes seen in humans. We tested the hypothesis that change in mPFC glutamate after an acute stressor would be moderated by behavioral factors related to social subordination in a manner similar to that previously observed in humans. Seventeen adult female rhesus monkeys (Macaca mulatta, 13-23 yrs.) were observed over ten weeks to collect behavioral data and then received two MRS scans. The first scan occurred after acute stress manipulation involving relocation and isolation. The second control scan occurred after acclimation to the new location. As expected, we found that a behavioral measure of social subordination predicted an adaptive glutamate response such that animals experiencing more submissive behavior asymmetry (a behavioral measure related to social subordination) exhibited an attenuated glutamate response to the acute stressor. These data establish the use of MRS to measure the adaptive glutamate stress in non-human primates and will help further our understanding of the neurobiology of stress adaptation.

Chronic psychosocial stress links to increased risk of psychiatric disorders. Human Magnetic Resonance Spectroscopy (MRS) found glutamate levels in medial prefrontal cortex (mPFC) post-acute stress exposure adapt to recent chronic stress levels. We sought to back-translate this glutamate stress response adaptation to rhesus macaques, whose societies are maintained by dominance, enforced by agonistic interactions, and result in phenotypes seen in chronically stressed humans. We tested the hypothesis that change in mPFC glutamate after acute stressor would be moderated by sociobehavioral factors.

Somatosensory deficits are seen in most stroke patients and can improve during recovery from stroke as reported previously. However, the underlining mechanism of the recovery remains poorly understood. The nonhuman primate (NHP) model of stroke has demonstrated similar functional connectivity evolution as seen in stroke patients and the secondary somatosensory cortex (S2) in NHPs is recognized as a direct extrapolation of somatosensory cortex in human. In the present study, the progressive function alteration of S2 and its relationship with the cerebral blood flow (CBF) and T2 changes in the monkey brain during acute stroke were examined and evaluated.

BACKGROUND:The oxygen extraction fraction (OEF) indicates the brain's oxygen consumption and can be estimated by using the quantitative susceptibility mapping (QSM) MRI technique. Recent studies have suggested that OEF alteration following stroke is associated with the viability of at-risk tissue. In the present study, the temporal evolution of OEF in the monkey brain during acute stroke was investigated using QSM.METHODS:Ischemic stroke was induced in adult rhesus monkeys (n = 8) with permanent middle cerebral artery occlusion (pMCAO) by using an interventional approach. Diffusion-, T2-, and T2*-weighted images were conducted on day 0, day 2, and day 4 post-stroke using a clinical 3T scanner. Progressive changes in magnetic susceptibility and OEF, along with their correlations with the transverse relaxation rates and diffusion indices, were examined.RESULTS:The magnetic susceptibility and OEF in injured gray matter of the brain significantly increased during the hyperacute phase, and then decreased significantly on day 2 and day 4. Moreover, the temporal changes of OEF in gray matter were moderately correlated with mean diffusivity (MD) (r = 0.52; p = 0.046) from day 0 to day 4. Magnetic susceptibility in white matter progressively increased (from negative values to near zero) during acute stroke, and significant increases were seen on day 2 (p = 0.08) and day 4 (p = 0.003) when white matter was significantly degenerated. However, significant reduction of OEF in white matter was not seen until day 4 post-stroke.CONCLUSION:The preliminary results demonstrate that QSM-derived OEF is a robust approach to examine the progressive changes of gray matter in the ischemic brain from the hyperacute phase to the subacute phase of stroke. The changes of OEF in gray matter were more prominent than those in white matter following stroke insult. The findings suggest that QSM-derived OEF may provide complementary information for understanding the neuropathology of the brain tissue following stroke and predicting stroke outcomes.

Background: Somatosensory deficits are frequently seen in acute stroke patients and may recover over time and affect functional outcome. However, the underlying mechanism of function recovery remains poorly understood. In the present study, progressive function alteration of the secondary somatosensory cortex (S2) and its relationship with regional perfusion and neurological outcome were examined using a monkey model of stroke. Methods and materials: Rhesus monkeys (n = 4) were induced with permanent middle cerebral artery occlusion (pMCAo). Resting-state functional MRI, dynamic susceptibility contrast perfusion MRI, diffusion-weighted, T1 and T2 weighted images were collected before surgery and at 4-6, 48, and 96 h post stroke on a 3T scanner. Progressive changes of relative functional connectivity (FC), cerebral blood flow (CBF), and CBF/Tmax (Time to Maximum) of affected S2 regions were evaluated. Neurological deficits were assessed using the Spetzler approach. Results: Ischemic lesion was evidently seen in the MCA territory including S2 in each monkey. Relative FC of injured S2 regions decreased substantially following stroke. Spetzler scores dropped substantially at 24 h post stroke but slightly recovered from Day 2 to Day 4. Relative FC progressively increased from 6 to 48 and 96 h post stroke and correlated significantly with relative CBFand CBF/Tmax changes. Conclusion: The present study revealed the progressive alteration of function connectivity in S2 during acute stroke. The preliminary results suggested the function recovery might start couple days post occlusion and collateral circulation might play a key role in the recovery of somatosensory function after stroke insult. The relative function connectivity in S2 may provide additional information for prediction of functional outcome in stroke patients.

With the ultimate goal of developing a more representative animal model of Alzheimer's disease (AD), two female amyloid-β-(Aβ) precursor protein-transgenic (APPtg) rhesus monkeys were generated by lentiviral transduction of the APP gene into rhesus oocytes, followed by in vitro fertilization and embryo transfer. The APP-transgene included the AD-associated Swedish K670N/M671L and Indiana V717F mutations (APPSWE/IND) regulated by the human polyubiquitin-C promoter. Overexpression of APP was confirmed in lymphocytes and brain tissue. Upon sacrifice at 10 years of age, one of the monkeys had developed Aβ plaques and cerebral Aβ-amyloid angiopathy in the occipital, parietal, and caudal temporal neocortices. The induction of Aβ deposition more than a decade prior to its usual emergence in the rhesus monkey supports the feasibility of creating a transgenic nonhuman primate model for mechanistic analyses and preclinical testing of treatments for Alzheimer's disease and cerebrovascular amyloidosis.

Background:The corticospinal tract (CST) is a major tract for motor function. It can be impaired by stroke. Its degeneration is associated with stroke outcome. Diffusion tensor imaging (DTI) tractography plays an important role in assessing fiber bundle integrity. However, it is limited in detecting crossing fibers in the brain. The crossing fiber angular resolution of intra-voxel structure (CFARI) algorithm shows potential to resolve complex fibers in the brain. The objective of the present study was to improve delineation of CST pathways in monkey brains scanned by conventional DTI.Methods:Healthy rhesus monkeys were scanned by diffusion MRI with 128 diffusion encoding directions to evaluate the CFARI algorithm. Four monkeys with ischemic occlusion were also scanned with DTI (b = 1000 s/mm2, 30 diffusion directions) at 6, 48, and 96 hours post stroke. CST fibers were reconstructed with DTI and CFARI-based tractography and evaluated. A two-way repeated MANOVA was used to determine significances of changes in DTI indices, tract number, and volumes of the CST between hemispheres or post-stroke time points.Results:CFARI algorithm revealed substantially more fibers originated from the ventral premotor cortex in healthy and stroke monkey brains than DTI tractography. In addition, CFARI showed better sensitivity in detecting CST abnormality than DTI tractography following stroke.Conclusion:CFARI significantly improved delineation of the CST in the brain scanned by DTI with 30 gradient directions. It showed better sensitivity in detecting abnormity of the CST following stroke. Preliminary results suggest that CFARI could facilitate prediction of function outcomes after stroke.

Background:Investigation of stroke lesion has mostly focused on grey matter (GM) in previous studies and white matter (WM) degeneration during acute stroke is understudied. In the present study, monkeys were utilized to investigate the alterations of GM and WM in the brain following ischemic occlusion using diffusion tensor imaging (DTI).Methods:Permanent middle cerebral artery occlusion (pMCAO) was induced in rhesus monkeys (n=6) with an interventional approach. Serial DTI was conducted on a clinical 3T in the hyperacute phase (2-6 hours), 48, and 96 hours post occlusion. Regions of interest in GM and WM of lesion areas were selected for data analysis.Results:Mean diffusivity (MD), radial diffusivity (RD), and axial Diffusivity (AD) in WM decreased substantially during hyperacute stroke, as similar as those seen in GM. No obvious fractional anasotropy (FA) changes were seen in GM and WM during hyper acute phase. until 48 hours post stroke when significant fiber losses were oberved also. Pseudo-normalization of MD, AD, and RD was seen at 96 hours. Pathological changes of WM and GM were observed in ischemic areas at 8, 48, and 96 hours post stroke. Relative changes of MD, AD and RD of WM were correlated negatively with infarction volumes at 6 hours post stroke.Conclusion:The present study revealed the microstructural changes in gray matter and white matter of monkey brains during acute stroke by using DTI. The preliminary results suggest axial and radial diffusivity (AD and RD) may be sensitive surrogate markers to assess specific microstructural changes in white matter during hyper-acute stroke.

Amitriptyline is a tricyclic antidepressant commonly prescribed in humans for pain and sleep disorders and in non-human primates for self-injurious behaviors. Here, we report a clinical case on the teratogenic effect of maternal-fetal amitriptyline exposure.

Objective: Alfaxalone has been used increasingly in biomedical research and veterinary medicine of large animals in recent years. However, its effects on the cerebral blood flow (CBF) physiology and intrinsic neuronal activity of anesthetized brains remain poorly understood. Methods: Four healthy adult rhesus monkeys were anesthetized initially with alfaxalone (0.125 mg/kg/min) or ketamine (1.6 mg/kg/min) for 50 min, then administrated with 0.8% isoflurane for 60 min. Heart rates, breathing beats, and blood pressures were continuously monitored. CBF data were collected using pseudo-continuous arterial spin-labeling (pCASL) MRI technique and rsfMRI data were collected using single-shot EPI sequence for each anesthetic. Results: Both the heart rates and mean arterial pressure (MAP) remained more stable during alfaxalone infusion than those during ketamine administration. Alfaxalone reduced CBF substantially compared to ketamine anesthesia (grey matter, 65 22 vs. 179 +/- 38 ml/100g/min, p<0.001; white matter, 14 +/- 7 vs. 26 +/- 6 ml/100g/min, p < 0.05); In addition, CBF increase was seen in all selected cortical and subcortical regions of alfaxalone-pretreated monkey brains during isoflurane exposure, very different from the findings in isoflurane-exposed monkeys pretreated with ketamine. Also, alfaxalone showed suppression effects on functional connectivity of the monkey brain similar to ketamine. Conclusion: Alfaxalone showed strong suppression effects on CBF of the monkey brain.The residual effect of alfaxalone on CBF of isoflurane-exposed brains was evident and monotonous in all the examined brain regions when used as induction agent for inhalational anesthesia. In particular, alfaxalone showed similar suppression effect on intrinsic neuronal activity of the brain in comparison with ketamine. These findings suggest alfaxalone can be a good alternative to veterinary anesthesia in neuroimaging examination of large animal models. However, its effects on CBF and functional connectivity should be considered.

BACKGROUND:Diffusion MRI (dMRI) data acquisition protocols are well-established on modern high-field clinical scanners for human studies. However, these protocols are not suitable for the chimpanzee (or other large-brained mammals) because of its substantial difference in head geometry and brain volume compared with humans. Therefore, an optimal dMRI data acquisition protocol dedicated to chimpanzee neuroimaging is needed.METHODS:A multi-shot (4 segments) double spin-echo echo-planar imaging (MS-EPI) sequence and a single-shot double spin-echo EPI (SS-EPI) sequence were optimized separately for in vivo dMRI data acquisition of chimpanzees using a clinical 3T scanner. Correction for severe susceptibility-induced image distortion and signal drop-off of the chimpanzee brain was performed and evaluated using FSL software. DTI indices in different brain regions and probabilistic tractography were compared. A separate DTI data set from n=34 chimpanzees (13 to 56 years old) was collected using the optimal protocol. Age-related changes in diffusivity indices of optic nerve fibers were evaluated.RESULTS:The SS-EPI sequence acquired dMRI data of the chimpanzee brain with approximately doubled the SNR as the MS-EPI sequence given the same scan time. The quality of white matter fiber tracking from the SS-EPI data was much higher than that from MS-EPI data. However, quantitative analysis of DTI indices showed no difference in most ROIs between the SS-EPI and MS-EPI sequences. The progressive evolution of diffusivity indices of optic nerves indicated mild changes in fiber bundles of chimpanzees aged 40 years and above.CONCLUSION:The single-shot EPI-based acquisition protocol provided better image quality of dMRI for chimpanzee brains and is recommended for in vivo dMRI study or clinical diagnosis of chimpanzees (or other large animals) using a clinical scanner. Also, the tendency of FA decrease or diffusivity increase in the optic nerve of aged chimpanzees was seen but did not show significant age-related changes, suggesting aging may have less impact on optic nerve fiber integrity of chimpanzees, in contrast to previous results for both macaque monkeys and humans.

The dorsolateral prefrontal cortex (DLPFC) and ventral lateral prefrontal cortex (VLPFC) play critical but different roles in working memory (WM) processes. Resting-state functional MRI (rs-fMRI) was employed to investigate the effects of neonatal hippocampal lesions on the functional connectivity (FC) between the hippocampus (H) and the DLPFC and VLPFC and its relation to WM performance in adult monkeys. Adult rhesus monkeys with neonatal H lesions (Neo-H, n = 5) and age- and gender-matched sham-operated monkeys (Neo-C, n = 5) were scanned around 10 years of age. The FC of H-DLPFC and H-VLPFC in Neo-H monkeys was significantly altered as compared to controls, but also switched from being positive in the Neo-C to negative in the Neo-H. In addition, the altered magnitude of FC between right H and bilateral DLPFC was significantly associated with the extent of the hippocampal lesions. In particular, the effects of neonatal hippocampal lesion on FC appeared to be selective to the left hemisphere of the brain (i.e. asymmetric in the two hemispheres). Finally, FC between H and DLPFC correlated with WM task performance on the SU-DNMS and the Obj-SO tasks for the control animals, but only with the H-VLPFC and SU-DNMS task for the Neo-H animals. In conclusion, the present rsfMRI study revealed that the neonatal hippocampal lesions significantly but differently altered the integrity in the functional connectivity of H-DLPFC and H-VLPFC. The similarities between the behavioral, cognitive and neural alterations in Neo-H monkeys and Schizophrenia (SZ) patients provide a strong translational model to develop new therapeutic tools for SZ.

BACKGROUND:Multi-compartment diffusion models such as Neurite Orientation Dispersion and Density Imaging (NODDI) have been increasingly used for diffusion MRI (dMRI) data processing in biomedical research. However, those models usually require multiple HARDI shells that may increase scanning duration substantially, and their application can be hindered in uncooperative patients (like infants) accordingly. Also, it is highly expected that the same dataset can be explored with multiple diffusion models for retrieving complementary information. METHODS:Multiple gradient-encoding schemes which consisted of 4-6 shells, moderate b-values (bmax =1,500 or 2,000 s/mm2), and 32-80 gradient directions were explored. The corresponding time of acquisition (TA) for a single scan ranged from 3 to 8 minutes respectively. The dMRI protocols were tested on macaque monkeys using a 3T clinical setting. The data were analysed using both NODDI and diffusion basic spectrum imaging (DBSI) models. RESULTS:The maps of orientation dispersion index (ODI) and CSF were consistent across the 4-6 shell sampling schemes. However, the corresponding intra-cellular volume fraction (ICVF) maps showed reduced pixel counts [1,100±98 (80 directions) vs. 806±70 (32 directions), one slice] in white matter when fewer gradient directions or lower b-value was applied. The hindered diffusion and CSF ratio maps were comparable across these sampling schemes. The maps of restricted diffusion ratio varied across the schemes. However, its mean ratios (0.23±0.02 vs. 0.22±0.01) and pixel counts (1,540±70 vs. 1,510±38, one slice) between the schemes of 80 and 32 directions with b=2,000 s/mm2 were comparable. CONCLUSIONS:The present study reports a fast multi-shell dMRI data acquisition and processing strategy which allows for obtaining complementary information about microstructural alteration and inflammation from a single dMRI data set with both NODDI and DBSI models. The proposed approach may be particularly useful for characterizing the neurodegenerative disorders in uncooperative patients like children or acute stroke patients in which brain injury is associated with inflammation.

Background: Previous Diffusion Tensor Imaging (DTI) studies have demonstrated the temporal evolution of stroke injury in grey matter and white matter can be characterized by DTI indices. However, it still remains not fully understood how the DTI indices of white matter are altered progressively during the hyperacute (first 6 hours) and acute stage of stroke (≤ 1 week). In the present study, DTI was employed to characterize the temporal evolution of infarction and white matter injury after stroke insult using a macaque model with permanent ischemic occlusion. Methods and materials: Permanent middle cerebral artery (MCA) occlusion was induced in rhesus monkeys (n=4, 10-21 years old). The brain lesion was examined longitudinally with DTI during the hyperacute phase (2-6 hours, n=4), 48 hours (n=4) and 96 hours (n=3) post-occlusion. Results: Cortical infarction was seen in all animals. The Mean Diffusivity (MD) in lesion regions decreased substantially at the first time point (2 hours post stroke) (35%, p <0.05, compared to the contralateral side) and became pseudo-normalized at 96 hours. In contrast, evident FA reduction was seen at 48 hours (39%, p <0.10) post-stroke. MD reduction in white matter bundles of the lesion area was much less than that in the grey matter during the hyper-acute phase but significant change was observed 4 hours (4.2%, p < 0.05) post stroke . Also, MD pseudonormalisation was seen at 96 hours post stroke. There was a significant correlation between the temporal changes of MD in white matter bundles and those in whole lesion areas during the entire study period. Meanwhile, no obvious fractional anisotropy (FA) changes were seen during the hyper-acute phase in either the entire infarct region or white matter bundles. Significant FA alteration was observed in entire lesion areas and injured white matter bundles 48 and 96 hours post stroke. The stroke lesion in grey matter and white matter was validated by pathological findings. Conclusion: The temporal evolution of ischemic injury to the grey matter and white matter from 2 to 96 hours after stroke onset was characterized using a macaque model and DTI. Progressive MD changes in white matter bundles are seen from hyperacute phase to acute phase after permanent MCA occlusion and temporally correlated with the MD changes in entire infarction regions. MD reduction in white matter bundles is mild in comparison with that in the grey matter but significant and progressive, indicating it may be useful to detect early white matter degeneration after stroke.

An adult rhesus macaque developed seizures after the induction of ischemic stroke. Initially, on the day of surgery, a focal ischemic lesion was present exclusively in the right caudate nucleus. By 48 h after stroke induction, the lesion had extended into the putamen, when a seizure was observed. Our report highlights the temporal changes in infarction of unilateral basal ganglia after acute stroke and the accompanying clinical symptoms. This unusual case may provide additional information regarding the involvement of the basal ganglia in seizures, given that prior case reports and studies usually have not described the temporal and spatial evolution of the lesion before clinical symptoms emerge.

Object Isoflurane is a commonly used volatile anesthetic agent in clinical anesthesia and biomedical research. Prior study suggested the cerebral blood flow (CBF) and default mode network (DMN) could be changed after prolonged administration of isoflurane. The normal maintenance doses of isoflurane may vary from light (similar to 0.75%) to deep (similar to 1.5 or 2%) anesthesia. However, it is not clear how the duration effects are affected by the altered doses. The present study is aimed to examine if the duration effects are affected when isoflurane concentration is altered within normal maintenance doses. Materials and methods: Adult rhesus monkeys (n = 5, 8-12 years old, 8-10 kg) were anesthetized and maintained at isoflurane levels 0.89 +/- 0.03%, 1.05 +/- 0.12%, or 1.19 +/- 0.08%. CBF and DMN of monkeys were examined using arterial spin-labeling perfusion and resting state functional MRI techniques. Results: the functional connectivity (FC) in the dominant DMN (posterior cingulate cortex (PCC) to anterior cingulated cortex (ACC) or media prefrontal cortex (MPFC)) decreased substantially and similarly during 4-h administration of isoflurane at any given maintenance dosage. CBF changes varied with isoflurane dosage. At the low dose (similar to 0.89%), CBF decreased in most brain regions. In contrast, no obvious changes was seen in those regions (except for the subcortex) when higher doses of isoflurane were applied. Conclusion: FC in DMN was reduced substantially during prolonged administration of isoflurane. The FC reduction was not varying significantly with maintenance doses of isoflurane but the duration effect on CBF was dose-dependent. Such duration effects of isoflurane administration on DMN and CBF should be considered in the interpretation of the outcome in related neuroimaging studies of anesthetized subjects.

With the advent of parallel imaging MRI techniques, whole-body MRI is being increasingly used in clinical diagnosis. However, its application in preclinical research using large animals remains very limited. In the present study, the whole-body MRI techniques for adult macaque monkeys were explored using a conventional clinic 3T scanner. The T1, T2 anatomical images, and MR angiography of adult macaque whole bodies were illustrated. The preliminary results suggest whole-body MRI can be a robust tool to examine multiple organs of non-human primate (NHP) models from head to toe non-invasively and simultaneously using a conventional clinical setting. As NHPs are intensely used in biomedical research such as HIV/AIDS and vaccine discovery, whole body MRI techniques can have a wide range of applications in translational research using NHPs.

Non-human primates (NHPs) resemble most aspects of humans in brain physiology and anatomy and are excellent animal models for translational research in neuroscience, biomedical research and pharmaceutical development. Cerebral blood flow (CBF) offers essential physiological information of the brain to examine the abnormal functionality in NHP models with cerebral vascular diseases and neurological disorders or dementia. Arterial spin labeling (ASL) perfusion MRI techniques allow for high temporal and spatial CBF measurement and are intensively used in studies of animals and humans. In this article, current high-resolution ASL perfusion MRI techniques for quantitative evaluation of brain physiology and function in NHPs are described and their applications and limitation are discussed as well.