abstract: Opioid use rates and related deaths continue to be a public health crisis; while there are many contributing factors to opioid use disorders, criteria for diagnosis include problems related to social functioning. Previous research indicates that laboratory rats, which are frequently used as animal models of addiction-related behaviors, are capable of prosocial behavior. The following collection of studies were performed to determine the effects of heroin on prosocial behavior in rats, as well as the role of the insula in both self-administration of heroin and prosocial behaviors. All of the experiments were conducted utilizing an established model of prosocial behavior in rats in which a performing rat releases a cagemate from a restrainer. The occurrence of and latency to free the confined rat was recorded. After baseline rescuing behavior was established, rats were allowed to self-administer heroin (0.06 mg/kg/infusion i.v.), and subsequent experimental conditions were imposed. Experimental conditions, in a series of different studies, included comparing heroin reinforcers with sucrose, chemogenetically modulating the insular cortex (both stimulatory and inhibitory processes) and administering excitotoxic lesions in the insula. There were significant differences in saving behaviors between heroin and sucrose groups demonstrating an opioid induced loss of prosocial behavior. Modulating the insula chemogenetically resulted in some restoration of these opioid related deficits, and insular lesions did not significantly impact prosocial behaviors, however, there were significant differences between rates of heroin intake in lesioned animals versus non-lesioned controls. Taken together, these results demonstrate the deleterious effects of heroin on prosocial behaviors and offer further support for the role of the insula in both addiction and social constructs. / Dissertation/Thesis / Doctoral Dissertation Psychology 2020

Dense Deposit Disease (DDD) causes chronic renal dysfunction which progresses to end-stage renal disease in about half of patients within 10 years of diagnosis. Deficiency of and mutations in complement Factor H (CFH) are associated with the development of DDD, suggesting that dysregulation of the alternative pathway (AP) of the complement cascade is important in disease pathophysiology. Patients with DDD are studied to determine whether specific allele variants of the genes of the alternative pathway of the complement system segregate preferentially with the DDD. We have screened coding and intronic regions of genes of the complement system in DDD cases and controls using PCR, restriction digest and bidirectional sequencing. We have been able to identify novel mutations, allele variants and haplotypes in several genes of the complement system which are associated with the DDD phenotype based on statistical analyses. Since we have identified several genes associated with DDD, we have determined possible gene-gene interactions using computational analyses. We have found a strong synergistic interaction between polymorphisms in CFH and C3. To ascertain if the associated allele variants had a functional impact in the complement activity of an individual, we have obtained blood samples from normal individuals and measured AP complement activity and genotyped CFH and C3 for these samples. Association between AP activity and genotypes is analyzed using statistical methods. Significant association of CFH and C3 variants with AP complement activity has been observed. We also have generated a mice deficient of CFH and Factor D (CFD). CFH deficient mice develop renal pathology similar to DDD in humans. Renal function and complement activity have been determined in the double knockout in comparison to CFH deficient and CFD deficient mice. Results have shown that absence of Factor D can inhibit complement activation in CFH mice. Our data imply that DDD is a complex genetic disease and that genes of the AP complement system contribute to level of complement activity and the pathogenesis of DDD. With this study, we hope to develop an effective diagnosis and treatment plan for DDD patients.

No description available.

京都大学 / 0048 / 新制・課程博士 / 博士(人間・環境学) / 甲第21845号 / 人博第874号 / 新制||人||210(附属図書館) / 2018||人博||874(吉田南総合図書館) / 京都大学大学院人間・環境学研究科共生人間学専攻 / (主査)教授 石原 昭彦, 教授 久代 恵介, 教授 神﨑 素樹 / 学位規則第4条第1項該当 / Doctor of Human and Environmental Studies / Kyoto University / DGAM

<div>An abdominal aortic aneurysm (AAA) is a pathological dilation of the abdominal aorta, as defined by a 50% increase in diameter or a diameter greater than 3 cm. While typically asymptomatic, there is a risk that the AAA will rupture, causing massive hemorrhaging and high mortality rates. Thus, once detected, the clinician must choose between surveillance and elective surgical repair. The first option carries the risk of rupture; the second risks complications and graft failure. Currently, clinical metrics of rupture risk are dependent on diameter and growth rate. However, a number of studies have indicated that, although rupture risk does increase with increased diameter, there are also a large number of patients with aneurysms for which the diameter criteria is insufficient. There remains a strong need to 1) determine better estimates of rupture risk in order to accurately assess the need for surgery and 2) improve surgical treatment to reduce perioperative risk. </div><div><br></div><div>Herein, we use ultrasound in mice to address these two prevalent uncertainties in aneurysm development and treatment. First, we further develop a murine aneurysm model that forms large aneurysms with distal thrombus. To increase the applicability of the model, we modulate aneurysm growth by altering elastase concentration and lysyl oxidase inhibition. We show that initial elastase concentration impacts aneurysm size, which is driven in part by a change in the degree of initial degradation of the aortic wall. We also demonstrate that lysyl oxidase inhibition (via BAPN) remains necessary for expansion even after the initial aneurysm formation and that removal of the lysyl oxidase inhibitor effectively stops growth in this model. As a final point, we show that female mice develop larger aneurysms than the males using this model. Then, with the aim of improving surgical treatment options, we explore the patency of various design parameters involved in tissue-engineered vascular grafts. To do so, we assess the allowable parameter design space of murine textile arterial grafts, so as to lead to better selection of key design components. Overall, the findings in this thesis demonstrate the applicability of ultrasound in small animals to improve aneurysm diagnostic and treatment options.</div>

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease caused by the death of motor neurons in the spinal cord and brain. ALS is a genetically complex disease; diverse mutations cause motor neuron death by disrupting various interrelated pathways. To date, no therapy targeting a single factor can rescue motor neuron loss, nor is it known how or why sub-populations of motor neurons are particularly vulnerable in disease. Many studies have pointed to the Transforming Growth Factor Beta (TGF-𝝱) signaling superfamily as a modifier of disease in human patients and in animal models. Here, we have used the SOD1G93A model of ALS to investigate if and how TGF-𝝱 signaling in motor neurons changes pathology in these animals. In the first part of this study we characterize canonical TGF-𝝱 activation in motor neurons in SOD1G93A animals compared to controls. We have found that a vulnerable motor neuron subpopulation upregulates TGF-𝝱RII, a receptor necessary and unique to the classical arm of the TGF-𝝱 signaling family, in a disease dependent manner. Despite the upregulation of TGF-𝝱RII in these cells, there is not a corresponding activation of downstream canonical TGF-𝝱 effectors in diseased motor neurons. Through in vivo genetic manipulation we found that TGF-𝝱RII is dispensable in motor neurons, but that ablation of TGF-𝝱RI, a key receptor in multiple arms of the TGF-𝝱 superfamily, decreases motor neuron survival in SOD1G93A animals. To further understand how this manipulation changes TGF-𝝱 activation in motor neurons, we performed iterative indirect immunoflourescence imaging. We have identified that knocking out TGF-𝝱RI from motor neurons disrupts downstream canonical TGF-𝝱 activation in these cells. To identify how TGF-𝝱 signaling changes gene expression in these cells we have used Visium, a spatial RNAseq method, on lumbar spinal cords from these animals We have identified and are currently investigating potential downstream targets of TGF-𝝱 signaling in motor neurons in SOD1G93A animals. These data suggest that motor neurons rely on TGF-𝝱 signaling for survival in disease and that TGF-𝝱 signaling is important to the biology of a known vulnerable population of motor neurons.

Genomic modifications occur slowly across generations, whereas short-term epigenetic transgenerational inheritance of adaptive phenotypes may be immediately beneficial to large numbers of individuals, acting as a bridge for survival when adverse environments occur. In this study we used dietary exposure to crude oil as an example of an environmental stressor to assess its effects from the molecular to the organismal levels in piscine and avian animal models. In addition, we assessed the role of the parental exposures on their offspring F1 generation. The research developed in this dissertation has contributed to several areas of investigation including molecular biology, animal physiology, and evolutionary biology. The quantitative information from these studies may be utilized to supplement information regarding the proximate and ultimate effects of environmental stressors on fish and bird populations. Furthermore, this information may be used as additional support for understanding the conservation of the responses from the molecular to the whole organismal levels across the vertebrate taxa, as well as their implications for population survival and maintenance. Additionally, the zebrafish (Danio rerio), the Siamese fighting fish (Betta splendens) and the king quail (Coturnix chinensis) have proven to be excellent models to start building a strong basis for understanding the effects of environmental stressors and transgenerational epigenetic phenomena using a multi-level approach. Furthermore, as more raw data and information is discovered, the concatenation of development, organismal variation, epigenetics inheritance, natural selection, speciation and evolution is being slowly decrypted.

Les maladies ischémiques du myocarde sont actuellement la principale cause de mortalité dans les pays développés. Malgré la diminution des délais de reperfusion et l’amélioration de la prise en charge médicale, la mortalité de l’infarctus du myocarde reste stable. Le développement de nouvelles solutions thérapeutiques reste donc nécessaire. Parmi ces nouveaux traitements, les dérivés cellulaires d'origine cardiaque (obtenues après mise en culture de fragments de biopsies) semblent prometteurs. Ces cellules sont obtenues à partir de biopsies myocardiques qui mises en culture vont spontanément s’organiser en cluster multicellulaires tridimensionnels appelés cardiosphères. La modification des conditions de culture permet ensuite d’obtenir une population de cellules uniques appelés cellules dérivés des cardiosphères (CDCs). Ces CDCs sont capables de diminuer la taille de l’infarctus dans l’infarctus constitué (études précliniques et cliniques) ainsi que dans l’infarctus aigu (ischémie reperfusion). Cependant plusieurs études réalisées chez le petit animal suggèrent que 1. Les cardiosphères pourraient avoir un potentiel thérapeutique supérieur à celui des CDCs et 2. Les exosomes sécrétés par les CDCs semblent être les médiateurs de leur effet, et l’injection des exosomes seuls pourraient permettre d’obtenir le même effet. Néanmoins, aucune étude n’a jusqu’à présent évalué les cardiosphères et les exosomes dans un modèle pre-clinique de gros animal. L’objectif de cette thèse sera donc de comparer les effets des cardiosphères, des CDCs et des exosomes sécrétés par les CDCs dans des modèles porcins d’ischémie reperfusion et d’infarctus constitué.Méthodes : Dans un premier temps, la technique de délivrance du traitement, la dose optimale et la sécurité seront évaluées ; dans un 2e temps des études randomisées contre placebo seront réalisées pour évaluer l’efficacité. Le modèle animal utilisé sera un modèle porcin d’un infarctus du myocarde reperfusé réalisé par occlusion de l’artère interventriculaire antérieure (après la première diagonale) par un ballon d’angioplastie. Pour les études d’ischémie reperfusion, les animaux seront traités 30 minutes après la reperfusion et suivi pendant 48 heures. Pour les études dans l’infarctus constitué, les animaux seront traités 4 semaines après l’infarctus puis suivi 4 semaines supplémentaires. L’efficacité sera évaluée par histologie (étendue du no-reflow et taille de l’infarctus) pour l’ischémie reperfusion, et par IRM (taille de la cicatrice et fonction ventriculaire gauche) pour l’infarctus constitué.Résultats attendus : nous espérons confirmer les résultats obtenus chez le petit animal à savoir démontrer une efficacité de cardiosphères supérieure à celle des CDCs, et une efficacité des exosomes au moins égale à celles des CDCs mais avec un profil de sécurité (notamment immunologique) supérieur. / Background: Ischemic heart disease is the first cause of death in western countries. Despite early reperfusion and improvement of medical care, myocardial infarction (MI) mortality remains constant. Therefore new treatments are desirable. Among those new treatments, heart derived cells (obtained from cardiac explants) are promising. Those cells are grown from cardiac explants which, in culture, will spontaneously self-organized in three-dimensional multicellular cluster named cardiospheres. When replated in adherent surface, those cardiospheres will yield to a population of single cells named cardiosphere-derived cells (CDCs). Allogenic CDCs have been shown to decrease infarct size both in convalescent MI (pre-clinical and clinical studies) and in ischemia-reperfusion. However several small animal studies suggest that 1. Cardiospheres may be more potent than CDCs and 2. CDC-secreted exosomes are likely to be the mediator of CDC effect and their injection may recapitulate the effect of CDCs. Nevertheless, no study has assessed the efficacy of cardiospheres and CDC-derived exosomes in a relevant pre-clinical large-animal model. We aim to compare the effect of cardiospheres, CDCs and CDC-derived exosomes in pig models of convalescent MI (adverse remodeling) and acute MI (ischemia reperfusion).Methods: First, delivery, optimal dose and safety will be optimized. Then, randomized placebo-controlled study will be performed to assess efficacy. For all studies, MI will be performed by balloon occlusion of the left anterior descending artery after the 1st diagonal. For the acute studies, pigs will be treated 30 minutes after reperfusion and followed for 48 hours. For the chronic studies, pigs will be treated 4 weeks after MI and followed for 4 additional weeks. Efficacy will be evaluated by histology (no-reflow and infarct size) for the acute studies and by MRI (scar size and left ventricular function) for the chronic studies.Expected results: we expect to confirm the results obtained in small animal models. Efficacy of cardiospheres may be better than CDCs, and exosomes should be at least as effective as CDCs but with a more favorable safety profile (especially immunological).

Anxiety disorders are the most common class of mental disorders in the United States, and they both promote and exacerbate disorders of substance abuse. Mounting evidence of sex differences in the relationship between anxiety disorders and alcoholism supports the potential existence of an anxiety-dependent vulnerability to alcohol abuse in women compared with men. One potential point of overlap in the physiological systems involved in anxiety response and reward processing is the amygdala. Here, a model of chronic stress in rodents was employed to probe changes in the electrophysiological and biochemical properties of the amygdala at a post-stress baseline and during a post-stress first exposure to alcohol. Electrophysiological data revealed that neurons in the centromedial amygdala were more responsive to stimulation in the basolateral amygdala in females compared with males, but a history of chronic stress altered the female response to match that of males with or without a history of chronic stress. Protein analysis of postsynaptic glutamatergic receptor expression and phosphorylation in the amygdala did not indicate any differences based on sex or exposure to stress or alcohol. These data demonstrate a sex difference in stress-induced alterations in amygdala circuitry and indicate a potential role for this circuitry in the comorbidity of anxiety disorders and alcoholism.

No description available.