In isolated skeletal muscle arterioles, increasing transmural pressure causes an increase in constriction. This active myogenic response varies with age and fiber type. Increased transmural pressure activates both Ca2+-activated (KCa) potassium channels and voltage-dependent (Kv) potassium channels; these channels have a role in the negativefeedback pathways that modulate depolarization and myogenic constriction. We tested the hypothesis that increased KCa channel and Kv channel activity contribute to reduced myogenic responsiveness in skeletal muscle arterioles of aged rats. 1A arterioles were isolated from soleus, an oxidative muscle, and superficial gastrocnemius, a glycolytic muscle, of young (4 mos) and aged (24 mos) Fischer 344 rats. Myogenic responses were assessed by increasing intraluminal pressure (0-140 cm H2O) in increments of 20cm H2O. Vasoconstrictor response were determined in response to increasing concentrations of the KCa channel blocker, charybdotoxin (CTX; 10-10 to 10-7 M) and the Kv channel blocker, 4-Aminopyridine (4-AP; 10-5 to 10-2 M). To determine the role of potassium channels in modulating the myogenic response, cannulated arterioles from soleus and gastrocnemius were incubated with CTX (50 nM) and 4-AP (5mM) for 15 minutes prior to evaluation of the myogenic response. Increased Kv channel activity contributes to reduced myogenic constriction in soleus and gastrocnemius muscle arterioles from aged rats. In soleus muscle arterioles, KCa channel activity opposes myogenic tone in young but not old rats. In gastrocnemius muscle arterioles, treatment with CTX did not eliminate age-related differences in the myogenic response, and the KCa channel contribution to myogenic tone was, in fact, greater arterioles from young as compared to old rats. Kv channels contribute to greater myogenic constriction in soleus arterioles, KCa channels appear to be more active in gastrocnemius muscle arterioles as compared to soleus muscle arterioles. Therefore Kv and KCa channels are tonically active in skeletal muscle arterioles, contributing to a hyperpolarizing force that opposes myogenic constriction. Furthermore, increased Kv channel activity contributes to the age-related reduction of myogenic constriction in soleus and gastrocnemius muscle arterioles.

No description available.

Mounting evidence suggests that the normal biomechanical state of arteries may include a nearly equibiaxial intramural stress, and that arteries tend to undergo rapid and dramatic remodeling when perturbed from this normal state. Technical developments in the early 1980s and late 1990s enabled in vitro and ex vivo studies, respectively, of isolated perfused microvessels, and it is clear that they share many similarities in behavior with arteries. To date, however, there has been no systematic study of the effects of biaxial loading on the biomechanical behavior of arterioles. In this project, we describe a modification to a prior in vitro arteriole test system that allowed us to investigate the role of altered axial stretch on the passive, myogenic, and fully contracted biaxial behavior of isolated rat cremaster arterioles. We show that axial stretches from 85% to 110% of normal values induce modest changes in the measured circumferential and axial stress-stretch behavior and similarly in traditional measures of distensibility and myogenic index. Nevertheless, altered axial stretch has a dramatic affect on the biaxial state of stress and it appears that near equibiaxial stress occur at axial stretches larger than those used previously. Whereas this finding will not affect prior estimates of material and functional behavior, it may have important implications for the design of long-term ex vivo and in vivo studies wherein vessel growth and remodeling are critical.

Mounting evidence suggests that the normal biomechanical state of arteries may include a nearly equibiaxial intramural stress, and that arteries tend to undergo rapid and dramatic remodeling when perturbed from this normal state. Technical developments in the early 1980s and late 1990s enabled in vitro and ex vivo studies, respectively, of isolated perfused microvessels, and it is clear that they share many similarities in behavior with arteries. To date, however, there has been no systematic study of the effects of biaxial loading on the biomechanical behavior of arterioles. In this project, we describe a modification to a prior in vitro arteriole test system that allowed us to investigate the role of altered axial stretch on the passive, myogenic, and fully contracted biaxial behavior of isolated rat cremaster arterioles. We show that axial stretches from 85% to 110% of normal values induce modest changes in the measured circumferential and axial stress-stretch behavior and similarly in traditional measures of distensibility and myogenic index. Nevertheless, altered axial stretch has a dramatic affect on the biaxial state of stress and it appears that near equibiaxial stress occur at axial stretches larger than those used previously. Whereas this finding will not affect prior estimates of material and functional behavior, it may have important implications for the design of long-term ex vivo and in vivo studies wherein vessel growth and remodeling are critical.

In normal tissues, blood supply is closely matched to tissue demand for wide ranges of oxygen demand and arterial pressure. This suggests that multiple mechanisms regulate blood flow. Theoretical models can be used to analyze these interacting mechanisms. One proposed mechanism for metabolic flow regulation involves the saturation-dependent release of ATP by red blood cells, which triggers an upstream conducted response signal and arteriolar vasodilation. To analyze this mechanism, oxygen and ATP levels are calculated along a flow pathway of seven representative segments, including two vasoactive arteriolar segments. The conducted response signal is dependent on ATP concentration. Arteriolar tone depends on the conducted response signal, local wall shear stress and wall tension. Arteriolar diameters are calculated based on vascular smooth muscle mechanics. The model can account for increases in perfusion consistent with experimental findings at low and moderate oxygen consumption rates despite the opposing effects of the myogenic and shear-dependent responses. Autoregulation, the maintenance of nearly constant blood flow as arterial pressure varies, is assessed in the presence or absence of the myogenic, shear-dependent and/or metabolic responses. The model results indicate that the combined effects of myogenic and metabolic regulation overcome the vasodilatory effect of the shear-dependent response to generate autoregulatory behavior. Capillary recruitment has been shown to increase the capacity for oxygen delivery during exercise. In the model, capillary density is assumed to depend on small arteriole diameter. The model predicts a significant increase in the range over which perfusion can be regulated when recruitment is included. Oscillations in diameter and tone are predicted under certain conditions, suggesting a novel mechanism for vasomotion. The conditions that give rise to oscillations are analyzed. It is shown that the appearance of oscillations depends in a complex way on a number of system parameters. In summary, the theoretical model provides a quantitative assessment of the myogenic, shear-dependent and metabolic responses that affect blood flow regulation and identifies a role for capillary recruitment and vasomotion in the control of blood flow.

Integrins are transmembrane heterodimeric proteins that link extracellular matrix (ECM) to cytoskeleton and have been shown to function as mechanotransducers in non-muscle cells. Synthetic integrin-binding peptide triggers Ca2+ mobilization and contraction in vascular smooth muscle cells (VSMCs) from rat afferent arteriole, indicating that interactions between ECM and integrins modulate vascular tone. RGD, an integrin binding peptide, triggered contraction in cultured VSMCs as observed by Electric Cell-Substrate Impedance Sensing technique. To examine whether integrins transduce extracellular mechanical stress into intracellular Ca2+ signaling events in VSMCs, unidirectional mechanical force was applied to freshly isolated renal VSMCs through paramagnetic beads coated with fibronectin (FN, natural ligand of α5β1 integrin in VSMCs). Pulling of fibronectin-coated beads with electromagnet triggered Ca2+ sparks, followed by global Ca2+ mobilization. Paramagnetic beads coated with low-density lipoprotein (LDL), whose receptors are not linked to cytoskeleton, were minimally effective in triggering Ca2+ sparks and global Ca2+ mobilization. Pre-incubation with ryanodine, cytochalasin-D, or colchicine substantially reduced the occurrence of Ca2+ sparks triggered by fibronectin-coated beads. Binding of VSMCs with antibodies specific to the extracellular domains of alpha5 and beta1 integrins triggered Ca2+ sparks simulating the effects of fibronectin-coated beads. Anti-β2- integrin antibody served as the negative control. Traction force microscopy studies showed that only the force transduced via integrins could potentially trigger cytoskeletal remodeling in cultured VSMCs. Atomic force microscopy revealed a significant increase in surface roughness in VSMCs when treated with RGD peptide though there was no difference in the maximum deflection of the force curves. Pre-incubation of microperfused afferent arterioles with ryanodine or integrin specific binding peptide inhibited pressure-induced myogenic constriction. In conclusion, integrins transduce mechanical force into intracellular Ca2+ signaling events in renal VSMCs. Integrin-mediated mechanotransduction is probably involved in myogenic response of afferent arterioles. Thus, integrins can potentially act as sensors for myogenic response phenomenon and affect the autoregulatory mechanism in the vasculature.

L’atteinte de la fonction endothéliale représente une phase précoce de l’athérosclérose, un stade où les patients sont généralement asymptomatiques. Il existe donc un intérêt certain à détecter la dysfonction endothéliale. Nous avons développé une technique de mesure des variations de flot artériel au niveau des membres supérieurs, basée sur la spectroscopie proche infrarouge (NIRS). Cette approche permettrait d’étudier le niveau d’atteinte vasculaire et probablement de quantifier le degré de dysfonction endothéliale périphérique lors d’une hyperémie réactive. L'expérience a été exécutée sur deux cohortes de 13 et de 15 patients et a été comparée à la pléthysmographie par jauge de contrainte (SGP) qui est considérée comme une méthode de référence. Par la suite, nous avons caractérisé la réponse endothéliale par modélisation de la courbe hyperémique du flot artériel. Des études préliminaires avaient démontré que la réponse hyperémique adoptait majoritairement une forme bi-modale. Nous avons tenté de séparer les composantes endothéliales-dépendantes et endothéliales-indépendantes de l’hyperémie. La quantification des deux composantes de la réaction hyperémique permet de calculer un indice de la ‘santé’ du système endothélial local. Cet indice est nommé le ηfactor. Les résultats montrent une forte corrélation des mesures de flots entre la technique développée et la méthode de référence (r=0.91). Nous avons conclu que NIRS est une approche précise pour la mesure non-invasive du flot artériel. Nous avons obtenu une bonne répétabilité (ICC = 0.9313) pour le ηfactor indiquant sa robustesse. Cependant des études supplémentaires sont nécessaires pour valider la valeur de diagnostic du facteur défini. Mots clés: hyperémie réactive, réponse myogénique, oxyde nitrique, athérosclérose, spectroscopie proche infrarouge / Atherosclerotic diseases are mainly caused by coronary and peripheral blood vessel disorders. Endothelial dysfunction represents an early phase in these diseases, when patients are generally asymptomatic. We developed a technique, based on near infrared spectroscopy (NIRS), for measurement of arterial blood flow variations in limbs during reactive hyperemia. The technique allows the study of the level of vascular impairment and probably quantifying the level of endothelial dysfunction at peripheral arteries. The experiment was performed on two cohorts of 13 and 15 patients and was compared to strain gauge plethysmography (SGP) which is considered as gold standard. Afterward, we characterized endothelial reaction during reactive hyperemia through blood flow variations by modeling the hyperemic curve. Preliminary studies have shown that the hyperemic response generally adopts a bimodal form. The first peak was attributed to myogenic reaction that is endothelial independent and the second one to local endothelial cells reaction. The quantification of the two hyperemic response components makes it possible to calculate an index of ‘health’ for local endothelial cells, named ηfactor. The results showed a strong correlation (r = 0.91) of blood flow measurements between the developed method and the gold standard. We concluded that NIRS is a precise technique for non-invasive measurement of blood flow. Moreover, we found a high repeatability (ICC = 0.9313) of the ηfactor in repeated measurements indicating its robustness. Nonetheless, more studies are required to validate the diagnosis value of the defined factor. Key words: reactive hyperemia, myogenic response, endothelial dependent vasodilatation, nitric oxide, atherosclerosis, near infrared spectroscopy (NIRS)

Malfunction of the system which regulates the bloodflow in the brain is a major cause of stroke and dementia, costing many lives and many billions of pounds each year in the UK alone. This regulatory system, known as cerebral autoregulation, has been the subject of much experimental and mathematical investigation yet our understanding of it is still quite limited. One area in which our understanding is particularly lacking is that of the role of nitric oxide, understood to be a potent vasodilator. The interactions of nitric oxide with the better understood myogenic response remain un-modelled and poorly understood. In this thesis we present a novel model of the arteriolar control mechanism, comprising a mixture of well-established and new models of individual processes, brought together for the first time. We show that this model is capable of reproducing experimentally observed behaviour very closely and go on to investigate its stability in the context of the vasculature of the whole brain. In conclusion we find that nitric oxide, although it plays a central role in determining equilibrium vessel radius, is unimportant to the dynamics of the system and its responses to variation in arterial blood pressure. We also find that the stability of the system is very sensitive to the dynamics of Ca²⁺ within the muscle cell, and that self-sustaining Ca2+ waves are not necessary to cause whole-vessel radius oscillations consistent with vasomotion.

A insuficiência cardíaca (IC) é uma síndrome que cursa com sintomas como intolerância ao exercício e sinais como o edema periférico (EP). Prejuízo na constrição miogênica (CM) acarreta em aumento da pressão hidrostática capilar e EP. Sabendo dos efeitos benéficos do treinamento físico (TF) sobre o sistema cardiovascular na IC, hipotetizou-se que o TF aeróbio é eficaz em restaurar o prejuízo na CM em artérias tibiais caudais (ATC) e o EP induzido pela IC. A IC induziu aumento no volume da pata o qual foi revertido pelo TF. A CM em ATC foi atenuada nos ICs, e o TF restaurou a mesma por vias como ROCK e razão SERCA-2/Fosfolambano, aumentando à cinética e a sensibilidade ao Ca2+. O TF restaurou o aumento no diâmetro interno (Di) observado nas ATC dos IC. Conclui-se, que as ATCs de ratos com IC apresentam prejuízo da CM e remodelamento eutrófico para fora os quais foram restaurados pelo TF. Os ajustes na CM induzidos pelo TF estão associados a uma melhora na mobilização do Ca2+i e na sensibilização das proteínas contráteis. / Heart failure (HF) is a syndrome with symptoms as exercise intolerance and signs as peripheral edema (PE). Impairment of myogenic constriction (MC) leads to enhancement of capillary hydrostatic pressure and PE. Knowing about the beneficial effects of physical training (PT) on cardiovascular system of HF, we hypothesize that PT is efficient in restore the impaired MC of caudal tibial artery (CTA) and PE induced by HF. HF increases paw volume, which was reversed by PT. The MC of CTA was blunted in HF, and PT restored it by pathway such as ROCK and SERCA-2/Phospholamban ratio, that increases Ca2+ kinetics and sensitization. PT restores the enhanced internal diameter (Di) observed in CTA of HF. Taken together, the CTAs of HF rats present an impairment of MC and eutrophic outward remodelling which are restored by PT. The PT adjustments observed in MC are associated with an improvement in Ca2+ mobilization and contractile protein sensitization.

L’atteinte de la fonction endothéliale représente une phase précoce de l’athérosclérose, un stade où les patients sont généralement asymptomatiques. Il existe donc un intérêt certain à détecter la dysfonction endothéliale. Nous avons développé une technique de mesure des variations de flot artériel au niveau des membres supérieurs, basée sur la spectroscopie proche infrarouge (NIRS). Cette approche permettrait d’étudier le niveau d’atteinte vasculaire et probablement de quantifier le degré de dysfonction endothéliale périphérique lors d’une hyperémie réactive. L'expérience a été exécutée sur deux cohortes de 13 et de 15 patients et a été comparée à la pléthysmographie par jauge de contrainte (SGP) qui est considérée comme une méthode de référence. Par la suite, nous avons caractérisé la réponse endothéliale par modélisation de la courbe hyperémique du flot artériel. Des études préliminaires avaient démontré que la réponse hyperémique adoptait majoritairement une forme bi-modale. Nous avons tenté de séparer les composantes endothéliales-dépendantes et endothéliales-indépendantes de l’hyperémie. La quantification des deux composantes de la réaction hyperémique permet de calculer un indice de la ‘santé’ du système endothélial local. Cet indice est nommé le ηfactor. Les résultats montrent une forte corrélation des mesures de flots entre la technique développée et la méthode de référence (r=0.91). Nous avons conclu que NIRS est une approche précise pour la mesure non-invasive du flot artériel. Nous avons obtenu une bonne répétabilité (ICC = 0.9313) pour le ηfactor indiquant sa robustesse. Cependant des études supplémentaires sont nécessaires pour valider la valeur de diagnostic du facteur défini. Mots clés: hyperémie réactive, réponse myogénique, oxyde nitrique, athérosclérose, spectroscopie proche infrarouge / Atherosclerotic diseases are mainly caused by coronary and peripheral blood vessel disorders. Endothelial dysfunction represents an early phase in these diseases, when patients are generally asymptomatic. We developed a technique, based on near infrared spectroscopy (NIRS), for measurement of arterial blood flow variations in limbs during reactive hyperemia. The technique allows the study of the level of vascular impairment and probably quantifying the level of endothelial dysfunction at peripheral arteries. The experiment was performed on two cohorts of 13 and 15 patients and was compared to strain gauge plethysmography (SGP) which is considered as gold standard. Afterward, we characterized endothelial reaction during reactive hyperemia through blood flow variations by modeling the hyperemic curve. Preliminary studies have shown that the hyperemic response generally adopts a bimodal form. The first peak was attributed to myogenic reaction that is endothelial independent and the second one to local endothelial cells reaction. The quantification of the two hyperemic response components makes it possible to calculate an index of ‘health’ for local endothelial cells, named ηfactor. The results showed a strong correlation (r = 0.91) of blood flow measurements between the developed method and the gold standard. We concluded that NIRS is a precise technique for non-invasive measurement of blood flow. Moreover, we found a high repeatability (ICC = 0.9313) of the ηfactor in repeated measurements indicating its robustness. Nonetheless, more studies are required to validate the diagnosis value of the defined factor. Key words: reactive hyperemia, myogenic response, endothelial dependent vasodilatation, nitric oxide, atherosclerosis, near infrared spectroscopy (NIRS)