Spatial Frequency-Based Analysis of Mean Red Blood Cell Speed in Single Microvessels: Investigation of Microvascular Perfusion in Rat Cerebral Cortex

BACKGROUND: Our previous study has shown that prenatal exposure to X-ray irradiation causes cerebral hypo-perfusion during the postnatal development of central nervous system (CNS). However, the source of the hypo-perfusion and its impact on the CNS development remains unclear. The present study developed an automatic analysis method to determine the mean red blood cell (RBC) speed through single microvessels imaged with two-photon microscopy in the cerebral cortex of rats prenatally exposed to X-ray irradiation (1.5 Gy). METHODOLOGY/PRINCIPAL FINDINGS: We obtained a mean RBC speed (0.9±0.6 mm/sec) that ranged from 0.2 to 4.4 mm/sec from 121 vessels in the radiation-exposed rats, which was about 40% lower than that of normal rats that were not exposed. These results were then compared with the conventional method for monitoring microvascular perfusion using the arteriovenous transit time (AVTT) determined by tracking fluorescent markers. A significant increase in the AVTT was observed in the exposed rats (1.9±0.6 sec) as compared to the age-matched non-exposed rats (1.2±0.3 sec). The results indicate that parenchyma capillary blood velocity in the exposed rats was approximately 37% lower than in non-exposed rats. CONCLUSIONS/SIGNIFICANCE: The algorithm presented is simple and robust relative to monitoring individual RBC speeds, which is superior in terms of noise tolerance and computation time. The demonstrative results show that the method developed in this study for determining the mean RBC speed in the spatial frequency domain was consistent with the conventional transit time method

Complex-valued analysis of arterial spin labeling–based functional magnetic resonance imaging signals

Cerebral blood flow-dependent phase differences between tagged and control arterial spin labeling images are reported. A biophysical model is presented to explain the vascular origin of this difference. Arterial spin labeling data indicated that the phase difference is largest when the arterial component of the signals is preserved but is greatly reduced as the arterial contribution is suppressed by postinversion delays or flow-crushing gradients. Arterial vasculature imaging by saturation data of activation and hypercapnia conditions showed increases in phase accompanying blood flow increases. An arterial spin labeling functional magnetic resonance imaging study yielded significant activation by magnitude-only, phase-only, and complex analyses when preserving the whole arterial spin labeling signal. After suppression of the arterial signal by postinversion delays, magnitude-only and complex models yielded similar activation levels, but the phase-only model detected nearly no activation. When flow crushers were used for arterial suppression, magnitude-only activation was slightly lower and fluctuations in phase were dramatically higher than when postinversion delays were used. Although the complex analysis performed did not improve detection, a simulation study indicated that the complex-valued activation model exhibits combined magnitude and phase detection power and thus maximizes sensitivity under ideal conditions. This suggests that, as arterial spin labeling imaging and image correction methods develop, the complex-valued detection model may become helpful in signal detection. Magn Reson Med, 2009. © 2009 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/64527/1/22106_ftp.pd

OCT methods for capillary velocimetry

To date, two main categories of OCT techniques have been described for imaging hemodynamics: Doppler OCT and OCT angiography. Doppler OCT can measure axial velocity profiles and flow in arteries and veins, while OCT angiography can determine vascular morphology, tone, and presence or absence of red blood cell (RBC) perfusion. However, neither method can quantify RBC velocity in capillaries, where RBC flow is typically transverse to the probe beam and single-file. Here, we describe new methods that potentially address these limitations. Firstly, we describe a complex-valued OCT signal in terms of a static scattering component, dynamic scattering component, and noise. Secondly, we propose that the time scale of random fluctuations in the dynamic scattering component are related to red blood cell velocity. Analysis was performed along the slow axis of repeated B-scans to parallelize measurements. We correlate our purported velocity measurements against two-photon microscopy measurements of RBC velocity, and investigate changes during hypercapnia. Finally, we image the ischemic stroke penumbra during distal middle cerebral artery occlusion (dMCAO), where OCT velocimetry methods provide additional insight that is not afforded by either Doppler OCT or OCT angiography

Pericyte-mediated regulation of capillary diameter: a component of neurovascular coupling in health and disease

Because regional blood flow increases in association with the increased metabolic demand generated by localised increases in neural activity, functional imaging researchers often assume that changes in blood flow are an accurate read-out of changes in underlying neural activity. An understanding of the mechanisms that link changes in neural activity to changes in blood flow is crucial for assessing the validity of this assumption, and for understanding the processes that can go wrong during disease states such as ischaemic stroke. Many studies have investigated the mechanisms of neurovascular regulation in arterioles but other evidence suggests that blood flow regulation can also occur in capillaries, because of the presence of contractile cells, pericytes, on the capillary wall. Here we review the evidence that pericytes can modulate capillary diameter in response to neuronal activity and assess the likely importance of neurovascular regulation at the capillary level for functional imaging experiments. We also discuss evidence suggesting that pericytes are particularly sensitive to damage during pathological insults such as ischaemia, Alzheimer’s disease and diabetic retinopathy, and consider the potential impact that pericyte dysfunction might have on the development of therapeutic interventions and on the interpretation of functional imaging data in these disorders

Imagerie multimodale des corrélats vasculaires du vieillissement cérébral

RÉSUMÉ Plusieurs décennies de recherche ont permis de démontrer que le vieillissement a des effets sur une multitudes de composantes du cerveau. En particulier, des preuves s'accumulent à l'effet qu'il existe un lien entre le fonctionnement du cerveau au niveau cognitif et la santé du système vasculaire, notamment le débit sanguin cérébral (DSC) qui diminue avec l'âge, ainsi que la santé cardiorespiratoire qui pourrait corréler à la performance cognitive selon certaines études. Plusieurs techniques d'imagerie cérébrale couramment utilisées en recherche, telles que le signal dépendant du niveau d'oxygénation du sang en imagerie par résonance magnétique (BOLDIRM), se basent sur les corrélats vasculaires de l'activité des neurones. Cela en fait des outils propices pour l'étude des effets vasculaires du vieillissement, qui influencent directement les signaux mesurés. Cette thèse a utilisé plusieurs techniques d'imagerie cérébrale basées sur l'hémodynamique pour étudier les effets du vieillissement sur le cerveau à différentes échelles spatiales, chez l'humain et dans un modèle animal chez le rat. Dans un premier temps, la microscopie biphotonique a été utilisée pour mesurer la vitesse des globules rouges, le diamètre et la densité des capillaires ainsi que l'hématocrite local dans près de 1000 capillaires chez 12 rats Long-Evans jeunes (3 mois) et 12 rats âgés (24 mois) anesthésiés. Il a été mesuré que la vitesse des globules rouges et le diamètre étaient plus élevés dans les capillaires de rats âgés (par 48 et 7% respectivement), tandis que l'hématocrite et la densité volumiques des capillaires étaient plus faibles (par 32 et 20%). Ces résultats suggèrent que la diminution du DSC avec l'âge serait surtout attribuable à une baisse de densité vasculaire. En second lieu, l'IRM et la spectroscopie résolue en temps de vol ont permis de mesurer le débit, l'oxygénation (sO2) et la concentration totale d'hémoglobine (HbT) dans les cerveaux d'humains jeunes (18-30 ans) et âgés (62-72 ans), en plus de la réponse à une tâche cognitive de Stroop en termes de BOLD et de DSC. La capacité cardiorespiratoire des sujets et été mesurée par un test de VO2max. Nous avons mesuré, dans le cortex préfrontal gauche sollicité par la tâche de Stroop, des valeurs plus faibles de DSC (par 19%), sO2 (par 6%) et HbT (par 21%) chez les sujets âgés. Dans le groupe âgé, les mesures de sO2 étaient corrélées à la performance cognitive dans la tâche Stroop ainsi qu'au VO2max, mais pas celles de DSC ni de HbT. Ces résultats suggèrent un effet protecteur de l'exercice physique sur la santé cognitive dans le vieillissement, dont les mécanismes seraient liés à une amélioration de l'oxygénation cérébrale. Enfin, les mêmes groupes de rats jeune et âgé ont été soumis à un stimulus vasodilatateur, l'hypercapnie, afin de mesurer la réponse hémodynamique à l'aide de----------ABSTRACT Several decades of research have demonstrated that aging affects a multitude of components in the brain. In particular, evidence is accumulating on the relation between brain function and vascular health, including cerebral blood flow (CBF), which decreases with age, and cardiopulmonary health which could correlate with cognitive performance according to some studies. Several brain imaging techniques commonly used in research, such as the blood oxygenation level dependent signal in magnetic resonance imaging (BOLD-MRI), are based on the vascular correlates of neural activity. This makes them suitable tools for the study of the vascular effects of aging, which directly influence the measured signals. This thesis used several imaging modalities based on hemodynamics to study the effects of aging on the brain at different spatial scales, in humans and in an animal model, the rat. Initially, two-photon microscopy was used to measure the velocity of red blood cells (RBCs), the diameter and the density of capillaries and the local hematocrit in nearly 1000 capillaries in 12 young (3 months-old) and 12 aged anesthetized Long-Evans rats (24 months-old). We measured higher RBCs velocity and diameter in the capillaries of aged rats (by 48 and 7 % respectively), while the hematocrit and volumetric capillary density were lower (by 32 and 20 %). These results suggest that the decrease in CBF with age is due primarily to a decrease in vascular density. Second, MRI and time-resolved spectroscopy were used to measure the CBF, oxygenation (sO2) and total hemoglobin concentration (HbT) in the brains of young (18-30 years-old) and elderly (62-72 years-old) humans, in addition to the response to a cognitive Stroop task in terms of BOLD and CBF. Cardiorespiratory fitness was measured by a VO2max test. In the left prefrontal cortex activated by the Stroop task, we measured lower values of CBF (by 19%), sO2 (by 6%) and HbT (by 21%) in the elderly. In the older group, measures of sO2 were correlated with Stroop task cognitive performance and with VO2max, while CBF and HbT were not. These results suggest a protective effect of physical activity on cognitive health in aging, mediated by an improvement in cerebral oxygenation. Finally, the same groups of young and old rats were subjected to a vasodilating stimulus, hypercapnia, for measuring the hemodynamic response with several imaging modalities. The data demonstrated a decrease in the hemodynamic response to hypercapnia in terms of CBF, HbT and HbO (oxygenated hemoglobin) in aged rats, suggesting decreased vascular reactivity. The vessels' compliance could also be reduced with age, as the ratio o

Aerobic exercise reverses aging-induced depth-dependent decline in cerebral microcirculation

Aging is a major risk factor for cognitive impairment. Aerobic exercise benefits brain function and may promote cognitive health in older adults. However, underlying biological mechanisms across cerebral gray and white matter are poorly understood. Selective vulnerability of the white matter to small vessel disease and a link between white matter health and cognitive function suggests a potential role for responses in deep cerebral microcirculation. Here, we tested whether aerobic exercise modulates cerebral microcirculatory changes induced by aging. To this end, we carried out a comprehensive quantitative examination of changes in cerebral microvascular physiology in cortical gray and subcortical white matter in mice (3-6 vs. 19-21 months old), and asked whether and how exercise may rescue age-induced deficits. In the sedentary group, aging caused a more severe decline in cerebral microvascular perfusion and oxygenation in deep (infragranular) cortical layers and subcortical white matter compared with superficial (supragranular) cortical layers. Five months of voluntary aerobic exercise partly renormalized microvascular perfusion and oxygenation in aged mice in a depth-dependent manner, and brought these spatial distributions closer to those of young adult sedentary mice. These microcirculatory effects were accompanied by an improvement in cognitive function. Our work demonstrates the selective vulnerability of the deep cortex and subcortical white matter to aging-induced decline in microcirculation, as well as the responsiveness of these regions to aerobic exercise

Spectral Domain-optical Coherence Tomography for the Assessment of Cerebrovascular Plasticity

Vascular pathologies represent the leading causes of mortality worldwide, accounting for 31% of all deaths in 2012. Cerebral hypoxia is a condition that often manifests as a result of these medical conditions. Remarkably, the nervous system has evolved mechanisms to compensate for oxygen deprivation. The dilation of existing vessels and the growth of new blood vessels are two prominent physiological responses to hypoxia, both of which play a critical role in maintaining cerebral homeostasis. More recently, exercise has been shown to induce a mild state of hypoxia in the brain, leading to several robust morphological changes within the cerebrovascular system (e.g., angiogenesis, vasodilation). Thus, exercise serves as a viable model for investigating hypoxia-induced adaptations. The present study introduces spectral domain optical coherence tomography (SD-OCT) as a novel technique for examining these micro-level changes in the rat motor cortex. SD-OCT produces high resolution, three-dimensional angiograms, and allows for moderately invasive imaging within the same animal at multiple time points. The independent effect of exercise training on cerebrovascular structure and function has never been explored using SD-OCT. Thus, the primary goal of this study was to determine the relative efficacy of SD-OCT utility. To validate this novel technology, we employed SD-OCT in the examination of exercise-dependent blood vessel growth, as well as real-time capillary dilation in response to a laboratory-induced condition of hypoxia (i.e., 10% oxygen). In addition, histology data was collected to provide comparative measures for statistical analyses. At the start of this investigation, animals were pseudo-randomly assigned to one of two groups: 26-week voluntary exercise (VX), or an inactive control (IC). Upon completing the exercise treatment, animals were anesthetized and prepared for imaging. Vascular anatomy and blood velocity data was captured during three experimental conditions: [1] normal oxygen baseline, [2] hypoxia – 10% oxygen, and [3] normoxia, return to baseline. A two-way analysis of variance revealed a significant difference in total blood vessel density between treatment groups, independent of condition. That is, VX animals had a greater density of blood vessels in the scanned region of interest when compared to IC. These findings were confirmed using unbiased stereology techniques to analyze tissue in the scanned region of interest. Furthermore, statistical analyses revealed a significant increase in small arteriole diameter in both VX and IC animals. However, the dilation captured by SD-OCT was significantly greater in VX animals when compared to IC. In sum, exercise induces potent adaptations that promote greater flexibility during hypoxia. Moreover, these micro-level changes can be effectively probed using SD-OCT

Basic and Clinical Understanding of Microcirculation

Microcirculation is key to providing enough nutrition and oxygen from head to toe. This is possible only through an extensive network of blood vessels spread around the body. Effect of microcirculation abnormalities stretch beyond one’s comprehension. The effects could be felt at any age, from the foetal life to the adulthood. The chapters present in this book describe how these abnormalities could lead to diseases such as atherosclerosis, thrombosis, diabetes, hypertension. Disorders of microcirculation could be related to the structural and/or functional damage to the inner lining of the blood vessels. Early identification of these disorders could benefit many ailments including cardiovascular and cerebrovascular diseases such as heart attack and stroke