When considering the array of terrestrial locomotor behaviors, bipedalism is a particularly rare way of moving about the landscape. In fact, humans are the only obligate terrestrial mammalian bipeds. Therefore, understanding both how and why it evolved is particularly intriguing. However, there is debate over why the evolution of bipedalism occurred and there is a large gap in knowledge for the mechanisms that underpin the evolution of these adaptive morphologies. One complicating factor for sorting out which models best explain how our hominin ancestors became bipedal is that they all rely on the same set of traits. Moreover, many of the traits that are thought to be diagnostic of bipedalism are only linked by association and have not been experimentally tested. That is, they do not appear in non-human primates and other quadrupeds. Therefore, addressing why the evolution of bipedalism occurred requires understanding the adaptive significance of traits linked with bipedalism. In this dissertation, I use an experimental approach employing both human and animal models to explore links between morphology and behavior and to tease apart the adaptive significance of particular traits. For the human portion of the dissertation, I use an inverse dynamics approach (estimating muscle forces from kinematic, kinetic, and anatomical data) to determine how modern human anatomy functions while walking using ape-like postures to clarify the links between morphology and energy costs in different mechanical regimes to determine the adaptive significance of postcranial anatomy. The results from this portion of the dissertation suggest that adopting different joint postures results in higher energy costs in humans due to an increase in active muscle volumes at the knee. These results lead to two conclusions important for understanding the evolution of human bipedalism. One is that human anatomy maintains low energy costs of walking in humans compared to chimpanzees regardless of lower limb postures. Second, the results suggest that erect trunk posture may be an important factor in reducing energy costs, therefore indicating that lumbar lordosis (the curvature of the lower spine) is important for reducing costs. For the animal portion of the dissertation, I use rats as a model for the quadrupedal-to-bipedal transition and experimentally induce bipedal posture and locomotion under a variety of loading conditions to determine if traits consistent with the evolution of bipedalism occur and under what conditions. This experimental design also has the ability to determine if there is a role for developmental plasticity in generating bipedal morphology to help answer the question how the evolution of bipedalism occurred. I find that inducing bipedal behaviors in a quadrupedal animal generates morphology consistent with human bipedal traits and that loading conditions have specific effects in different skeletal elements and at particular joints. I also find that there is a plausible role for developmental plasticity in generating adaptive bipedal morphology in the earliest hominins. Overall, the results from the experimental procedures in this dissertation were able to clarify links between behavior and bipedal morphology, demonstrate a plausible role for developmental plasticity in early adaptation to bipedal behavior in australopiths, determine the adaptive significance of human postcranial anatomy, and the ways in which postcranial anatomy reduces costs.

Molecular and genetic analysis of synaptic signaling in Drosophila has yielded many insights into nervous system development, properties of synaptic transmission, and how long-lasting changes in neurons occur. Synaptic signaling components required for synaptic transmission and pathways leading to nervous system plasticity are typically conserved from insects to humans. The role of proteins and genes in synaptic function in flies can be analyzed from the level of a single synapse to complex behaviors in the whole organism. Because of a fully sequenced genome and the ease of mutagenesis in flies, genetic screens have been useful in identifying novel regulators of synaptic transmission and long-term memory.In flies, conditional mutations affecting synaptic transmission at nerve terminals often lead to temperature sensitive paralysis. In a screen for mutations that interact with Drosophila shibirets mutants, the stoned gene was identified as a regulator of synaptic vesicle cycling. Stoned encodes two neuronally expressed proteins, stonedA and B, which are required for synaptic vesicle recycling and normal synaptic transmission. However, the exact functions of the two stoned proteins are not fully understood. We investigate distinct roles of the stoned proteins here and show that stoned has a novel role in synaptic growth.Memory in flies can be divided into genetically distinct phases based on the requirement for protein synthesis and activation of the transcription factor CREB. Novel regulators of long-term olfactory avoidance memory were isolated in a mutant screen in flies. Mutants in the Drosophila gene lk6, homologous to the translational regulator MNK, have defects in long-term olfactory avoidance memory. We find that lk6 is highly expressed in the fly nervous system, and is activated by and functions downstream of Ras/ERK signaling in fly neurons. Insights provided here from Drosophila add to the evidence that MNK may be the link between ERK signaling and the regulation of translation in long-term plasticity.Ultimately, understanding synaptic function has therapeutic potential to aid in alleviation of nervous system dysfunction. Insight into the molecular pathways underlying plasticity and long-term memory gained from studies in flies, mollusks, and rodents has been pivotal in the development of potential drugs to aid in memory deficits in humans.

Phenotypic plasticity, the ability of a genotype to express different phenotypes in different environments, allows organisms to cope with variation in resources and invade novel environments. Biologists have long been fascinated with the costs and tradeoffs that generate and maintain variation in plasticity, such as possible increases in brain size and delays in reproduction associated with the evolution of learning. However, the costs of plasticity vary: many studies have failed to find costs of plasticity, the degree of costs often vary with the system or environments considered, and many costs of plasticity are variable even within the lifetime of an individual. This research adopts a developmental perspective to predict the degree and incidence of costs of plasticity, using host learning in butterflies as a case study. Learning, a mechanism of plasticity that develops through a trial-and-error sampling process, should result in developmental costs and allocation of energy towards development (at the expense of reproduction). Furthermore, costs of learning should be less pronounced in environments for which organisms have innate biases and for learned traits underlain by short-term memory, relative to long-term memory (which requires more developmental re-structuring). This research found support for all three predictions across three levels of costs: behavioral costs, tissue costs, and fecundity trade-offs. Butterflies exhibited genetic variation in their ability to learn to recognize different colored hosts. Genotypes with higher proxies for long-term memory emerged with relatively larger neural investment and smaller reproductive investment. In contrast to these costs of long-term learning, proxies of short-term learning were only correlated with increased exploration of a range of possible resources (types of non-hosts) early in the host-learning process. Family-level costs of plasticity emerged from the ability to learn to locate a red host, for which butterflies do not have an innate bias. Costs of learning were also induced by learning itself: following exposure to novel (red) host environments, individual butterflies, regardless of genetic background, increased exploratory behavior, increased neural investment, and re-allocated energy away from reproduction towards other functions (e.g., flight). Considering developmental mechanisms helps to predict how costs will influence the evolution of learning and plasticity.

Reproductive traits are often thought of as fixed, genetically determined properties. However, such traits are often dynamic, exhibiting different expression patterns depending on context. Both internal state and external environment can have a strong effect on how traits are expressed. Variation in these factors across the lifetime of an individual should select for flexibility in trait expression, rather than fixation.My dissertation work examines how mating behavior and testes size respond to several previously unexplored contextual factors, using Rhagoletis juglandis, the walnut fly, as a model system. For mating behavior, I predicted that differences in female reproductive state (egg load) and experience with host resource would impact mating decisions. For testes size, I predicted that social environment (sex ratio) and changes in resource environment would determine testes size.Behavioral observations of flies showed that a large egg load increased the likelihood of copulation, while prior experience with host fruit decreased copulation time. These results are the first to distinguish effects of experience on physiological state from other effects of experience in the context of mating behavior.Manipulation of the sex ratio revealed that males develop larger testes when reared in an environment with many potential competitors. This is the first study to show that that allocation to a male reproductive organ can change depending on the sex ratio. My studies showed that resource environment is also important in determining testes investment patterns. When adult males are deprived of protein, they develop smaller testes. A stable isotope analysis of testes further confirms that resource environment is important for testes development. Males rely more on nitrogen derived at the larval stage than at the adult stage, but adult carbon sources are a large component of testes mass.In sum, this dissertation demonstrates the importance of context in the expression of reproductive traits. Recent research has shown that such traits can respond more dynamically to context than previously thought, but this area of research is young. My results help provide a greater understanding of the processes shaping the evolution of reproductive traits.

No description available.

Investigators have shown that elephants exhibit consistent individual differences in behavior by rating elephants using personality adjectives. These adjectives, however, are not based on pre-defined measurements of the behaviors performed. Instead, they are based on the observers’ interpretations of an animal’s behavioral patterns, therefore making them subject to observer bias. Furthermore, elephants have a capacity for learning; thus, they may alter their behavioral patterns over time. This behavioral plasticity in itself might be a way of measuring consistent behavioral differences among individuals. With this in mind, I approached elephant personality as a multivariate problem. I used behavioral observations collected from female elephants in Addo Elephant National Park, South Africa. Instead of grouping behaviors into subjective categories prior to analysis (as is often done in studies of elephant behavior), I used ordination methods to determine which correlations among behaviors were important for defining personality. Ordination methods were performed on matrices of the behavior data set and on subsets of behaviors for each age class. I calculated the angular differences among major axes of covariation from the ordinations of subsets to determine if the behaviors that defined personalities differed by age class. I also defined personalities by centroids (in multidimensional space) for non-metric multidimensional scaling (NMDS) scores of each individual and dispersion of NMDS scores for each individual as a measure of behavioral plasticity. I analyzed the effects of plasticity and age on personality of individual elephants using a non-parametric multivariate analysis of variance. Major axes of covariation were not well defined and therefore not useful in describing differences among groups. The interaction of age and behavioral plasticity did have a significant effect on the personalities of individuals as defined by ordination centroid scores. This suggests that incorporating plasticity may be a helpful measurement in quantifying consistent behavioral differences among individuals.

Primary visual cortex is a classic model to study experience-dependent brain plasticity. In early life, if one eye is deprived of normal vision, there can be a dramatic change in the ocular dominance of the striate cortex such that the large majority of neurons lose responsiveness to the deprived eye and, consequently, the ocular dominance distribution shifts in favor of the open eye. Interestingly, the visual experience dependent plasticity following monocular deprivation (MD) occurs during a transient developmental period, which is called the critical period. MD hardly induces ocular dominance plasticity beyond critical period. The mechanisms underlying ocular dominance plasticity during the critical period are not fully understood. It has been proposed that long-term depression (LTD) may underlie the loss of cortical neuronal responsiveness to the deprived eye. However, discordant results have been reported in terms of the role of LTD and LTP in visual plasticity due to the lack of specific blockers. Here we report the prevention of the normally-occurring ocular dominance (OD) shift to the open eye following MD by using a specific long-term depression (LTD) blocking peptide derived from the GluR2 subunit of the a-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptor (AMPAR). We were able to prevent the shift of OD to the open eye with systemic or local administration of the GluR2 peptide. Both electrophysiological and anatomical approaches were taken to demonstrate the peptide effect. Moreover, enhancing LTD with D-serine, a NMDA receptor co-agonist, brought back the ocular dominance plasticity in adult mice subject to four-day MD and, therefore, reopened the critical period. Our data indicate that LTD plays an essential role in visual plasticity during the critical period and the developmental regulation of LTD may account for the closure of critical period in adult. In an additional study, we have found anisomycin, a protein synthesis inhibitor, produces a time-dependent decline in the magnitude of the field EPSP (fEPSP) in mouse primary visual cortex and that this anisomycin-mediated fEPSP depression occludes NMDA receptor dependent LTD. In contrast, another two protein synthesis inhibitors, emetine and cycloheximide, have no effect either on baseline synaptic transmission and or on LTD. We propose that anisomycin-LTD might be mediated by p38 MAP kinase since anisomycin is also a potent activator of the P38/JNK MAPK pathway. In agreement with notion, the decline of the fEPSP caused by anisomycin can be rescued by the application of the P38 inhibitor SB203580, but not by the JNK inhibitor SP600125. The occlusion of LFS-LTD by anisomycin-induced fEPSP decline suggests that common mechanisms may be shared between the two forms of synaptic depression. Consistent with this view, bath application of the membrane permeant peptide discussed above, which specifically blocks regulated AMPA receptor endocytosis, thereby preventing the expression of LFS-LTD, prior to anisomycin treatment significantly reduced the anisomycin-induced decline of the fEPSP. In conclusion, this study indicates that anisomycin produces long-lasting depression of AMPA receptor-mediated synaptic transmission by activating P38 MAPK-mediated endocytosis of AMPA receptors in neonatal mouse visual cortex.

The automotive industry is adopting lightweight materials to improve emissions and fuel economy. Magnesium (Mg) alloys are the lightest of engineering metals, but work is required to assess their structural strength, especially for spot-welded applications. In the present research, fatigue behavior of magnesium spot-welds was characterized and compared with steel and aluminum spot-welds. A fatigue model was proposed to predict the failure location and crack initiation life in magnesium structures. The material under investigation, AZ31B-H24 Mg alloy, and its spot-welds were characterized from microstructural and mechanical perspectives. Microstructure and hardness of the base metal (BM) and different regions in the spot-welds were studied. Under cyclic loading, the BM had an asymmetric hysteresis loop. Cyclic behavior of magnesium spot-welds was measured using different specimen configurations, and the effect of geometrical factors on fatigue life was evaluated. A constitutive model was developed to model the asymmetric hardening behavior of wrought magnesium alloys under cyclic loading. An algorithm for numerical implementation of the proposed model was developed. The numerical formulation was programmed into a user material subroutine to run with the commercial finite element software Abaqus/Standard. The proposed model was verified by solving two problems with available solutions. A number of available fatigue models, as well as a new model proposed in this research were assessed by predicting fatigue life of magnesium spot-welds. The new model used a strain energy damage parameter. All models were evaluated by comparing the predicted and experimental fatigue lives for different Mg spot-welded specimens. The effect of considering the asymmetric hardening behavior of wrought magnesium alloys on the accuracy of the fatigue life prediction was not significant for the available experimental data. This was attributed to the limited experimental data on spot-welded specimens. The proposed material model and fatigue damage parameter were verified by simulating a real-life structure manufactured and fatigue tested by the US Automotive Materials Partnership. The results obtained from the proposed asymmetric model were compared with available symmetric simulation results and experimental data. The asymmetric material model along with the proposed damage parameter resulted in more accurate prediction of fatigue failure location and life.

Olfaction is vitally important to animals in all environments and is used to identify food, habitat, conspecifics and predators. Some odors, like pheromones or the pungent smell of spoiled foods, can trigger pre-existing behavioral responses that appear to require no learning. Most odors, however, are only attended to as a result of prior experience. It is believed that different types of odors are processed in different olfactory pathways in the forebrain. This thesis examines the relationship between innate and learned olfactory behaviors and the anatomy of the neural pathways that underlie them, using the zebrafish olfactory system as a model. I first characterized an appetitive olfactory behavior, which is displayed promptly by zebrafish when they encounter amino acid odors. A similar appetitive behavior can also be learned by the fish for another, initially neutral odorant, if it is repeatedly paired with food rewards. Zebrafish can therefore respond to, and learn to respond to certain odors. I then conducted an in-depth anatomical analysis of the structure and distribution of glomeruli in the zebrafish olfactory system. Glomeruli are spheroidal synaptic aggregates that organize and shape olfactory information that arrives in the brain. Throughout the development of zebrafish, I identified two distinct populations of glomeruli. One population consisted of 25 individually identifiable, anatomically stereotypic glomeruli that closely resembled specialized glomeruli in mammals and insects. These glomeruli were already formed during embryonic development and persisted in remarkably stable configurations throughout later developmental stages. I hypothesize that the 25 individually identifiable glomeruli constitute stable olfactory pathways (i.e., for innate olfactory behaviors). Most glomeruli, however, were anatomically variable and displayed different distributions within coarsely circumscribed regions in the zebrafish olfactory bulbs. The development of these glomeruli could be modified by sensory experience, suggesting that they may comprise plastic olfactory pathways that subserve the establishment of learned olfactory behaviors. Collectively my results show that innate and learned olfactory behaviors may indeed be represented in different olfactory pathways, and that these types of pathways may be located in both main and accessory olfactory systems.

The LIM-Kinase family of proteins (LIMK) plays an important role in actin dynamics through its regulation of ADF/cofilin. A subtype of LIMK, LIMK1, is mostly expressed in neuronal tissues with high levels in the mature synapse. Previous studies from the Zhen Ping Jia laboratory have shown that LIMK1-/- mice exhibit abnormal spine morphology as well as altered hippocampal synaptic plasticity. LIMK1 has been shown to interact with CREB during neuronal development (Yang et al., 2004). We propose that LIMK1 is able to phosphorylate CREB in response to a synaptic activity. We hypothesize that if LIMK1 activates CREB in mature neurons, then LIMK1 knockout mice will have decreased L-LTP and deficits in long-term memory. My results show that LIMK1 and CREB exist in a complex and are bound to each other in mature neurons. LIMK1-/- mice exhibit deficits in the late phase of long-term potentiation and specific deficits in long-term memory while short-term memory remains unaltered. Pharmacological activation of CREB attenuates the observed deficits in synaptic plasticity and long-term memory. These results show a potentially novel mechanism of CREB activation in response to synaptic activity. Moreover, using peptides to manipulate actin dynamics in LIMK1 lacking animals only has effects on early LTP and is not able to rescue the late phase LTP deficits found in LIMK1 -/- mice. These results indicate a specific role of LIMK1 long-term memory and synaptic plasticity through regulation of CREB and not through regulation of the actin cytoskeleton.