The normal aging process in humans is characterized by a number of hallmark changes including decreased white matter volume in the brain and accompanying cognitive decline. This is in contrast to neurodegenerative aging processes which involve acute pathology which results in neuronal cell death. Studying non-degenerative normal aging in humans can be difficult because of the high prevalence of neurodegenerative diseases in the population and other potentially confounding effects. Rhesus monkeys are an excellent model organism for the study of normal aging, as their aging process has been demonstrated to involve diminished white matter volume, but they do not suffer from neurodegenerative diseases such as Alzheimer's. In this study we seek to quantify levels of myelin degradation using confocal microscopy in regions of interest where it has been previously demonstrated that loss of white matter integrity results in lower levels of cognitive function across different treatment groups including aging monkeys, calorie restricted monkeys, and controls for calorie restricted monkeys. These areas include prefrontal white matter which is vital to executive function, the hippocampus which is integral to memory consolidation and the learning process, and finally the anterior, middle, and posterior cingulum bundle. The cingulum bundle contains a diverse variety of projections between cortical and subcortical regions, including but not limited to projections to and from the cingulate cortex which has been demonstrated to be vital for emotional processing, the limbic system, and a wide spectrum of other functions. We aim to quantify white matter degradation in these regions by using immunofluorescent tagging for healthy myelin basic protein (MBP) and degraded myelin basic protein (dMBP) and by measuring the colocalization between the two. For prefrontal white matter and hippocampus, we did not find significant differences in myelin degradation across treatment groups. In the cingulum bundle, however, we did find a significant effect of treatment on overall myelin damage throughout the bundle, and in particular we determined that there was a significant difference in colocalization in the anterior cingulum bundle between aging monkeys and control calorie restricted monkeys. Analysis of behavioral testing data yielded surprising results as we were unable to find a strong correlation between our measure for myelin degradation, and level of cognitive impairment. Our results indicate that there are likely differences in regional vulnerability to age related myelin damage across different white matter regions of the brain, however we would like to expand on this study to gain a more accurate understanding of how loss of white matter volume is distributed through the brain and the impact that has on cognitive outcomes.

Oocyte quality plays a critical role in establishment of pregnancies, embryo development, implantation and the health of offspring. The oocyte provides maternal factors necessary for the initial development of its embryo during the period of transcriptional silence. Despite the consistent increase in number of couples seeking assisted reproductive treatments, oocyte quality still remains as an obstacle in successful fertility treatments and the mechanisms governing the quality of oocyte are poorly understood. Among various factors that may potentially affect the quality of oocyte, the acquisition of oocyte developmental competence seems to mainly occur during the final stage of oocyte maturation. The correct temporal regulation of series of molecular events and the proper exchange of signals with surrounding follicular environment during this critical period will ensure the developmental competence of oocyte and its subsequent embryo. In order to identify molecular factors affecting oocyte quality, I have compared oocytes and cumulus cells of different qualities at a molecular level. I present in this thesis the pathways and molecules that may determine the developmental competence of oocyte as well as candidate molecular markers of oocyte and embryo quality. A cDNA microarray analysis was performed, comparing the transcriptomes of rhesus monkey MII oocytes of different qualities, high quality VVM oocytes and poor quality IVM oocytes. A small set of 59 Oocyte quality plays a critical role in establishment of pregnancies, embryo development, implantation and the health of offspring. The oocyte provides maternal factors necessary for the initial development of its embryo during the period of transcriptional silence. Despite the consistent increase in number of couples seeking assisted reproductive treatments, oocyte quality still remains as an obstacle in successful fertility treatments and the mechanisms governing the quality of oocyte are poorly understood. Among various factors that may potentially affect the quality of oocyte, the acquisition of oocyte developmental competence seems to mainly occur during the final stage of oocyte maturation. The correct temporal regulation of series of molecular events and the proper exchange of signals with surrounding follicular environment during this critical period will ensure the developmental competence of oocyte and its subsequent embryo. In order to identify molecular factors affecting oo was identified as differentially expressed between the two types of oocytes. These mRNAs are involved in steroid metabolism, cell-cell interactions, cellular homeostasis, cell adhesion, mRNA stability and translation. In addition, the overexpression of several imprinted genes in IVM oocytes were detected, indicating a possible loss of correct epigenetic programming during IVM. These results indicate that normal oocyte-somatic cell interactions may be disrupted during IVM and the interruptions of these interactions during the final phase of oocyte maturation may be the prime cause of reduced developmental competence of IVM oocytes. To elucidate oocyte quality factors linked to the cumulus cell phenotype, the transcriptomes of two types of rhesus monkey cumulus cells, IVM and VVM, were compared using a cDNA microarray analysis. In contrast to a relatively small difference between IVM and VVM oocytes, a large number of genes were differentially expressed between IVM and VVM rhesus cumulus cells. Moreover, a much larger number of differential mRNA expressions were observed comparing the transitions from pre-maturation cumulus cells to the IVM and VVM cumulus cells. The results from these array comparisons indicated that the cumulus cells may fail to achieve successfully normal gene regulation during IVM and thus make a remarkable amount of changes in gene expression to compensate for the loss. Numerous genes involved in lipid metabolism are incorrectly regulated during IVM, and the synthesis of sex hormones appears not suppressed during IVM. In addition, a panel of 24 cumulus cell markers of oocyte quality was identified. Genetic effects on oocyte quality were explored by comparing transcriptomes of oocytes obtained from two different inbred mouse strains, B6 and D2, and F1 hybrid. A clustering analysis and statistical tests showed that the transcriptome of F1 oocytes is more similar to the B6 transcriptome than to the D2 at both GV and MII stages. Also, comparison analyses of GV stage oocyte transcriptomes with MII oocyte transcriptomes from three different mouse strains indicated that the number of overdominance genes at the MII stage is bigger than the number of overdominance genes at the GV stage. In order to investigate how the genes gain the overdominance during the GV to MII transition, overdominance genes were categorized according to their mRNA expression patterns at GV and MII stages. The results showed that more than 80% of overdominance genes belong to one of the four major transition groups. The further evaluation of changes in array intensities from GV to MII stage transition revealed that F1 oocytes and inbred strain oocytes differentially regulate the mRNA abundance during oocyte maturation and that the differential regulation of mRNA abundance by the F1 genotype is responsible for the increase of the number of overdominance genes during maturation from GV stage to MII stage. A mRNA sequence analysis indicated that the gain of overdominant low in F1 mRNA expression pattern during maturation may be regulated by 3'UTR motif elements. The number of dominance genes also increase during GV to MII transition. At both GV and MII stages, there are more genes with B6 dominant mRNA expression pattern than those with D2 dominance pattern. Lipid metabolism, small molecule biochemistry and cell death are biofunctions overrepresented in both dominance and overdominance genes. In addition, `blebbing' was identified as a biofunction significantly downregulated in the F1 and B6 MII eggs, indicating that the cellular function may be involved in oocyte maturation. / Molecular Biology and Genetics / Accompanied by one .pdf file: YLEE_SupplementalTables.pdf

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2017-04-24T14:23:01Z No. of bitstreams: 1 2016 - Carlos Augusto dos Santos Sousa.pdf: 2340153 bytes, checksum: 5ef373f242c2c4700a9a9e55280bc62c (MD5) / Made available in DSpace on 2017-04-24T14:23:01Z (GMT). No. of bitstreams: 1 2016 - Carlos Augusto dos Santos Sousa.pdf: 2340153 bytes, checksum: 5ef373f242c2c4700a9a9e55280bc62c (MD5) Previous issue date: 2016-02-03 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Morphology studies provide knowledge that allow us to understand how animals interact with the natural environment or in captivity. In this context, the comparative anatomy of the formation of the brachial plexus awakens interest since the nineteenth century and remains one of the most intriguing topics of contemporary anatomy. The aim of this study was to describe the origin and the antimeric distribution of the brachial plexus nerves in Macaca mulatta, as well as the innervated muscles. Ten male rhesus monkeys (Macaca mulatta) were used, from the Non-human Primates? Breeding Department at the Laboratory Animals Breeding Centre (Cecal/Fiocruz), donated to the Animal Anatomy Department of the Rural Federal University of Rio de Janeiro (UFRRJ). The specimens were fixed in formaldehyde solution by infusion of 10% solution. They were subsequently wrapped in a low-density polythene container with 500 liters of formaldehyde 30% solution over a period of 12 months. After this period, they were washed in running water and subjected to X-ray examinations of the neck at the Small Animals Veterinary Hospital of the UFRRJ to characterize the number of cervical vertebrae. Then, they had both antimeres dissected aiming at the exposure of the origins and the nerves arising from the brachial plexus. Data were presented both in absolute frequency and in simple percentage. In 11 (55%) animals the resulting nerves were constituted by the connections between the ventral spinal branches C5, C6, C7, C8 and T1. In 5 (25%) animals, the participants roots were C4, C5, C6, C7, C8, T1 and T2. In 2 (10%) animals C5, C6, C7, C8, T1 and T2. In the other 2 (10%) animals the formation of the plexus was observed from C6, C7, C8, T1 and T2. The ventral branches formed three nerve trunks: cranial, middle and caudal. The suprascapular nerves, subscapular, axillary, musculocutaneous, radial, median, ulnar innervated the intrinsic muscles and the subclavian nerve innervated the thoracodorsal, medial cutaneous arm and forearm, long thoracic, cranial pectoral and caudal pectoral innervate extrinsic muscles. The results obtained in this study contribute to the comparative anatomy of primates and to the information for applied research, serving as basis for clinical and surgical procedures that uses this species as an animal model. / Estudos morfol?gicos fornecem conhecimentos que permitem entender o modo como os animais interagem com o ambiente natural ou em cativeiro. O objetivo desse estudo foi descrever a origem e a distribui??o antim?rica dos nervos do plexo braquial em Macaca mulatta, assim como dos m?sculos inervados. Foram utilizados 10 cad?veres de Macaca mulatta do sexo masculino, oriundos do Servi?o de Cria??o de Primatas N?o Humanos do Centro de Cria??o de Animais de Laborat?rio (Cecal/Fiocruz) doados a ?rea de Anatomia Animal da Universidade Federal Rural do Rio de Janeiro (UFRRJ). Os esp?cimes foram fixados com perfus?o de solu??o de formalde?do a 10%. Posteriormente, foram acondicionados em caixas de polietileno de baixa densidade com capacidade de 500 litros contendo solu??o de formalde?do a 30% por um per?odo de 12 meses. Ap?s este per?odo, foram lavados em ?gua corrente e submetidos a exames radiogr?ficos da regi?o cervical no Hospital Veterin?rio de Pequenos Animais da UFRRJ para a caracteriza??o do n?mero de v?rtebras cervicais. Em seguida, foram dissecados at? a exposi??o das origens e dos nervos oriundos do plexo braquial. Os dados foram representados em frequ?ncia absoluta e percentual simples. Em 11 (55%) os nervos resultantes foram constitu?dos das conex?es entre os ramos espinhais ventrais de C5, C6, C7, C8 e T1. Em 5 (25%) as ra?zes participantes foram C4, C5, C6, C7, C8, T1 e T2. Em 2 (10%) de C5, C6, C7, C8, T1 e T2. Em outros 2 (10%) verificamos a constitui??o do plexo a partir de C6, C7, C8, T1 e T2. Os ramos ventrais formaram tr?s troncos nervosos: cranial, m?dio e caudal. Os nervos supraescapular, subescapulares, axilar, musculocut?neo, radial, mediano, ulnar inervaram a musculatura intr?nseca e os nervos subcl?vios, toracodorsal, tor?cico longo, peitoral cranial e peitoral caudal inervaram a musculatura extr?nseca. Tamb?m foram registrados os nervos cut?neos oriundos do plexo braquial, sendo eles o nervo cut?neo medial do bra?o, nervo cut?neo medial do antebra?o e ramos para a musculatura cut?nea do tronco. Os dados descritos neste estudo contribuem para a anatomia comparada de primatas e fornecem informa??es para a pesquisa aplicada, servindo como base para procedimentos cl?nico-cir?rgicos em que venha a se utilizar esta esp?cie como modelo experimental.

Comparative research on event memory has typically focused on the binding of spatial and temporal information in memory, but much less is known about how animals remember information about the source of their memories (i.e., whether the event is something they performed themselves or whether they observed it). The purpose of the present study was to examine how rhesus monkeys (n = 8) and 3- to 4- year-old children (n = 20) remember this information along with other relevant event features (object identity, spatial location, temporal properties and contextual features) in working memory. In Experiment 1, rhesus monkeys completed five different delayed matching-to-sample tasks to assess independent encoding of these five event components. In Experiment 2, the monkeys either performed or observed an event and then had to respond to a randomly selected pair of memory tests used in the previous experiment. In Experiment 3, children were presented with the same memory task, but were given a brief demonstration to learn how to perform the task. Both children and monkeys responded to these tests using photos and shapes (for the identity and spatial tests) and icons (for the temporal, agency and context tests). The monkeys demonstrated significantly above-chance performance on the identity, spatial, temporal and agency tasks. The children were above chance on the one component the monkeys had difficulty with (context), but conversely demonstrated difficulty on the temporal memory test. There was evidence of feature integration in both monkeys and children. Specifically, the children were significantly more likely to respond correctly to the second memory test if they had also been correct on the first memory test. Two of five rhesus monkeys also showed this effect, indicating that for these individuals, the features were integrated in working memory. Implications of this research are discussed in relation to self-awareness and episodic memory research in children and nonhuman species.

Comparative research on event memory has typically focused on the binding of spatial and temporal information in memory, but much less is known about how animals remember information about the source of their memories (i.e., whether the event is something they performed themselves or whether they observed it). The purpose of the present study was to examine how rhesus monkeys (n = 8) and 3- to 4- year-old children (n = 20) remember this information along with other relevant event features (object identity, spatial location, temporal properties and contextual features) in working memory. In Experiment 1, rhesus monkeys completed five different delayed matching-to-sample tasks to assess independent encoding of these five event components. In Experiment 2, the monkeys either performed or observed an event and then had to respond to a randomly selected pair of memory tests used in the previous experiment. In Experiment 3, children were presented with the same memory task, but were given a brief demonstration to learn how to perform the task. Both children and monkeys responded to these tests using photos and shapes (for the identity and spatial tests) and icons (for the temporal, agency and context tests). The monkeys demonstrated significantly above-chance performance on the identity, spatial, temporal and agency tasks. The children were above chance on the one component the monkeys had difficulty with (context), but conversely demonstrated difficulty on the temporal memory test. There was evidence of feature integration in both monkeys and children. Specifically, the children were significantly more likely to respond correctly to the second memory test if they had also been correct on the first memory test. Two of five rhesus monkeys also showed this effect, indicating that for these individuals, the features were integrated in working memory. Implications of this research are discussed in relation to self-awareness and episodic memory research in children and nonhuman species.

Analysis of rhesus monkey alphoid DNA suggests that it arose by tandem duplication of an ancestral monomer unit followed by independent variation within two adjacent monomers (one becoming more divergent than the other) before their amplification as a dimer unit to produce tandem arrays. The rhesus monkey alphoid DNA is a tandemly repeated, 343-bp dimer; the consensus dimer is over 98% homologous to the alphoid dimers reported for baboon and bonnet monkey, 81% homologous to the African green monkey alpha monomer, and less than 70% homologous to the more divergent human alphoid DNAs. The consensus dimer consists of two wings (I and II, 172 and 171 bp, respectively) that are only 70% homologous to each other, but share seven regions of exact homology. These same regions are highly conserved among the consensus sequences of the other cercopithecid alphoid DNAs. The three alpha-protein binding sites reported for African green monkey alpha DNA by F. Strauss and A. Varshavsky (Cell 37: 889-901, 1984) occur in wings I and II, but with one site altered in wing I. Two cloned dimer segments are 98% homologous to the consensus, each containing 8 single-base-pair differences within the 343-bp segment. Surprisingly, 37% of these differences occur in regions that are evolutionarily conserved in the alphoid consensus sequences, including the alpha-protein binding sites. Sequence variation in this highly repetitive DNA family may produce unique nucleosomal architectures for different members of an alphoid array. These unique architectures may modulate the evolution of these repetitive DNAs and may produce unique centromeric characteristics in primate chromosomes.

Adopted individuals have an increased risk for a variety of psychopathological disorders. Studies of the effects adoption in humans are difficult to perform because of the difficulty separating genetic risk and treatment effects. This is a developmental study investigating the effects of at birth adoption using a nonhuman primate model. Three experimental paradigms were used to assess maternal treatment, stress-related behavior, and physiology late in infancy and again later in life. Rhesus monkeys were reared for their first six months of life by either their biological mother or an unrelated, lactating adult female. Adoptions occurred immediately following birth. At six months of age, both groups were exposed to four, 4-day mother-infant separations. Behavioral observations and plasma stress hormones were used to compare the two group's responses to the separation stressor. Maternal treatments were also compared. In a second experiment performed about three years later when subjects were adolescents or young adults, an unfamiliar intruder was placed outside their home pen and stress-related behavioral responses were again measured. In the third experiment, adolescent subjects were allowed free access to a sweetened alcohol solution and daily alcohol consumption was measured across 8-10 weeks. Analyses showed that adopted subjects exhibited more behavior withdrawal and higher ACTH during the Acute and Chronic phases of the separation than infants reared by their biological mothers. This persisted when subjects were again tested with an intruder stressor 1-3 years later, with adopted subjects still showing more behavioral withdrawal during the Intruder Challenge stressor. Adopted subjects also differ in their relationship with their mother, showing more independence at an early age in non-stressful environments. Paradoxically, alcohol intake was lower in adolescents raised by an adoptive mother. Differences in maternal treatment and mismatches in temperament between the adopted mother and her infant are potential mechanisms that lead to the increased stress and anxiety in subjects raised by an adopted mother.

Operant conditioning using positive reinforcement techniques has been used extensively in the management of nonhuman primates in both zoological and laboratory settings. Based on a large body of previous research that demonstrates the utility of such techniques in reducing stress, abnormal behavior, and aggression, this research project was intended to develop and test the usefulness of habituation and counter-conditioning techniques in reducing the fear-responses of singly-housed male rhesus macaques living in the laboratory environment. Additionally, we investigated the variable of temperament as it relates to the reduction of fear-responsivity and overall training success. Based on a Wilcoxon Matched-Pairs Sign Test, we found that animals exposed to desensitization training were significantly likely to show a reduction in the rate at which they engaged in cringing toward humans (exact significance = .016, one-tailed, N ties = 6), cringing in general (exact significance = .016, one-tailed, N ties = 6), and in stress-related behaviors (exact significance = .016, one-tailed, N ties = 6). Animals exposed to basic husbandry training or exposed to no training at all were not significantly likely to show a reduction in the rates of these behaviors. When these same behaviors were analyzed in terms of duration of behavior, desensitization-exposed animals were significantly likely to show reduction in the amount of time spent cringing toward humans (exact significance = .016, one-tailed, N ties = 6), but not in cringing behaviors in general or in stress-related behaviors. Neither the husbandry-exposed group nor the group exposed to no training showed a significant number of subjects exhibiting a reduction in duration of any of these behaviors. Additionally, initial temperament assessments were found to significantly predict the relative ability of subjects exposed to training to acquire trained behaviors such that animals generally ranked as more inhibited in terms of temperament also ranked as slower learners based on a Wilcoxon Matched-Pairs Signed-Ranks test, z = -.316, p = .752 (two-tailed). Results of this study could enhance both laboratory animal welfare and laboratory animal research, and could be a first step in developing techniques for reducing fearful behavior in rhesus monkeys in the laboratory environment.

Le suivi des changements dans les populations de cellules immunitaires tels que les lymphocytes, monocytes et cellules dendritiques (DC) au cours de maladies infectieuses comme le virus de l'immunodéficience humaine (VIH) chez l’homme ou son équivalent chez le singe (VIS) est crucial. Grâce aux récentes avancées technologiques en cytométrie en flux, il est maintenant possible de mesurer et d’analyser simultanément jusqu'à 14 paramètres individuels à l’échelon cellulaire. L'objectif de ce travail consiste en la mise au point de 2 panels multicouleurs de 12 anticorps permettant d'analyser simultanément les principales populations de cellules immunitaires, respectivement chez l’humain et le macaque rhésus. Au terme de ce travail, il est maintenant possible de mesurer précisément tous les principaux acteurs du système immunitaire, à savoir les lymphocytes T CD4+ et T CD8+, les lymphocytes B, les cellules NK et NKT, les sous-populations de monocytes, et toutes les sous-populations de cellules dendritiques connues à ce jour, en utilisant une approche multiparamétrique de cytométrie en flux. Ce protocole d’analyse est réalisé sur du sang total, il est rapide, il n’implique pas de technique d’isolation cellulaire, et requiert une quantité minimum de sang. De plus, l’analyse de chaque population cellulaire est plus précise grâce à une contamination minimum entre les populations séparées. L’intérêt de ce travail est d’étudier les interactions entre les différentes populations de cellules immunitaires durant l’infection par VIH chez l’homme, ou VIS chez le singe ou potentiellement d‘autres maladies, et en particulier de mieux comprendre le rôle important que les cellules dendritiques jouent dans la progression de ces maladies. / Monitoring changes in immune cell populations such as lymphocytes, monocytes and dendritic cells (DC) during infectious diseases like human immunodeficiency virus (HIV) or its counterpart in rhesus monkeys (SIV) is crucial. Thanks to recent technological advances in flow cytometry, it is now possible to measure and analyze simultaneously up to 14 individual parameters at the single cell level.The goal of this work is to develop 2 multicolor flow cytometry panels comprising of 12 antibodies, allowing measuring simultaneously the main immune cells population, respectively in humans and rhesus monkeys. After 2 years of development and optimization, we can now measure precisely all the main actors of the immune system, that is CD4+ and CD8+ T lymphocytes, B lymphocytes, NK and NKT cells, the 3 monocyte subsets, and all the dendritic cell subsets known today, by using a multicolor flow cytometry approach. This assay is done on whole blood, it is rapid to do, it does not involve a cell isolation technique, and it requires only a minimum amount of blood. Moreover, the analysis of each population is much more precise because of a minimum contamination between different cell populations. The advantage of this work is to study interactions between different cell populations of immune cells during HIV infection in humans, or SIV infection in monkeys, or potentially other diseases, and in particular to better understand the important role that dendritic cells might play in disease progression.

<p>My thesis examines how eugenicist and psychologist Robert M. Yerkes’ experimental intelligence research helped to situate the non-human primate as the ideal research subject for human betterment research in the twentieth century U.S. Yerkes believed that the primate was the ideal research subject to address questions of human betterment and social welfare, specifically best to create methods of evaluating the imagined threat of intellectual disability. While Yerkes has been studied extensively in the history of psychology, primatology, and eugenics, rarely have his separate contributions to these fields been placed in conversation with one another. Placing the primate at the center of Yerkes’ work allows for all three fields to engage with one another in a new perspective. By analyzing Yerkes’ publications about the Multiple-Choice Experiment within the context of the American eugenics’ movement, we can see how the primate came to hold a central position in U.S. scientific research, the advancement of human welfare and betterment, and as a means of defining what it means to be human. This story offers a glimpse into this longer process of how the primate came to occupy this position, but even a glimpse offers historians of the American eugenics’ movement new questions. What was the role of the non-human animal in the formulation of American eugenic theories? How have we historically used the natural world in our attempts to separate ourselves from it? And can we truly reconcile a history with eugenics if we continue to ignore the role of animals within it, they who today exist unquestionably within the status of the sub-human?</p>