The olfactory sensitivity of five male CD-1 mice (Mus musculus) for six amino acids was determined using an operant conditioning paradigm. All animals significantly distinguished dilutions as low as 0.01 mM L-cysteine, 3.3 mM L-methionine, 10 mM L-proline, 0.03 mM D-cysteine, 0.3 mM D-methionine and 10 mM D-proline from the odorless solvent, with individual animals displaying even lower detection thresholds. Among the three different L-forms of the amino acids the mice were most sensitive for cysteine and least sensitive for proline, and among the three D-forms the animals displayed a lower sensitivity for D-proline compared to D-cysteine and D-methionine. A comparison between the present data and results obtained with other species showed that the CD-1 mice displayed a higher sensitivity than human subjects and spider monkeys with three (L-Cysteine, D-cysteine and L-proline) of the six amino acids. Results from this report support the idea that the number of functional olfactory receptor genes is not suitable to predict a species’ olfactory sensitivity.

<p>The chemical cues that provide an olfactory portrait of mammalian individuals are in part detected by chemosensory receptors in the vomeronasal organ (VNO). By and large, the pertinent receptor-cue combinations used for olfactory communication are unidentified. Here we identify members of the formyl peptide receptor (FPR) family of G protein coupled receptors as candidate chemosensory receptors in the VNO of mice. We demonstrate that N-formylated mitochondrially encoded peptides presented by the major histocompatibility complex (MHC) molecule H2-M3 stimulate a subset of the VNO sensory neurons (VSNs). We show that one VNO localized FPR, Fpr-rs1, is differentially activated by strain specific variants of N-formylated peptides. We show that N-formylated peptides can function as chemosignals in a strain selective pregnancy block. We propose that this link between self-recognition peptides of the immune system and chemosensory pathways provides a possible molecular means to communicate the nature of an individual's maternal lineage or strain.</p> / Dissertation

<p>Altered dietary availability of the vital nutrient choline during early development leads to persistent changes in brain and behavior throughout adulthood. Prenatal choline supplementation during embryonic days (ED) 12-17 of the rodent gestation period enhances memory capacity and precision and hippocampal plasticity in adulthood, and protects against spatial learning and memory deficits shortly after excitotoxic seizures, whereas prenatal choline deficiency can compromise hippocampal memory and plasticity in adulthood. Recent evidence from our laboratory has determined that lifelong proliferation of newborn neurons in the adult hippocampus, a feature of adult hippocampal plasticity that has been implicated in some aspects of learning and memory, is modulated by early choline availability. Prenatal choline's effects on adult neurogenesis may be one mechanism for diet-induced cognitive changes throughout life and in response to injury, although little is known about the mechanisms underlying how prenatal choline alters adult neurogenesis or the neural mechanisms underlying prenatal choline supplementation's protection against cognitive deficits after seizures. To address these issues, the present set of experiments investigated how prenatal choline availability modulates specific properties of neurogenesis in the adult brain (in the intact brain and in response to injury), as well as hippocampal markers known to change in response to excitotoxin-induced seizures, and sought to relate changes in neurogenesis and in neuropathological markers following injury to changes in performance on spatial learning and memory tasks. Subjects in each experiment were adult offspring from rat dams that received either a control diet or diet supplemented with choline chloride or deficient of choline on ED 12-17. To measure neurogenesis, rats were given injections of the mitotic marker bromodeoxyurdine to label dividing cells in the hippocampus. Prenatal choline supplementation enhanced several properties of basal adult hippocampal neurogenesis (cell division and survival, neural stem/progenitor cell phenotype and proliferative capacity, trophic support), and this increase was associated with improvements in spatial working memory retention in a delayed-matching-to-place water maze task. In contrast, prenatal choline deficiency had little effect on basal adult hippocampal neurogenesis, and no effect on spatial memory performance. Prenatal choline supplementation also enhanced olfactory bulb neurogenesis without altering cell proliferation in the subventricular zone, while prenatal choline deficiency had no effect on either measure, showing for the first time that prenatal choline's effects on adult neurogenesis is similarly expressed in another distinct neurogenic region of the adult brain. Altered prenatal choline availability also modulated the hippocampal response to kainic acid-induced seizures where supplementation attenuated while deficiency had no effect on the injury-induced proliferative response of the dentate gyrus shortly after injury. Prenatal choline supplementation also attenuated other markers of hippocampal neuropathology shortly after seizures and promoted the long-term hippocampal recovery from seizures months after injury, including rescuing declines in adult hippocampal neurogenesis and in spatial memory performance in a standard water maze task. Taken together, these findings demonstrate a robust neuroprotective effect of prenatal choline supplementation that may be driven by enhanced adult hippocampal plasticity and trophic support prior to injury, and shed light on the mechanisms underlying how prenatal choline availability alters adult hippocampal neurogenesis, which may contribute to changes in memory capacity and precision both throughout life and following neural assault.</p> / Dissertation

No description available.

Das Geruchssystem ist in der Lage, mittels sogenannter kombinatorischer Kodierung einen hochdimensionalen Geruchsraum durch eine begrenzte Anzahl von olfaktorischen Rezeptorneuronen (ORN) abzutasten. Hierbei weisen verschiedene ORN-Klassen eine breite und gleichzeitig spezifische Geruchssensitivität auf, durch welche ein geruchsspezifisches Antwortmuster auf Populationen von Mitral-/Tufted Zellen (M/T) des bulbus olfactoris (OB) abgebildet wird. Neueren Untersuchungen zufolge sind diese Antwortmuster nicht notwendigerweise statisch, sondern enthalten Information in ihrer zeitlichen Entwicklung. Im OB von Larven des Krallenfrosches Xenopus laevis wurde herausgefunden, dass sowohl Geruchsidentität als auch -Konzentration besser vorhergesagt wird durch M/T Antwortlatenzmuster als durch durchschnittliche Feuerraten. Diese Arbeit befasst sich mit der Messung von ORN-Aktivität auf verschiedenen raumzeitlichen Skalen. Auf der Ebene von ORN Populationen wurde mit Hilfe von konfokaler Mikroskopie und [Ca2+] -sensitiven Fluoreszenzfarbstoffen untersucht, in wie weit Latenzmuster auftreten. Es wurde gezeigt, dass Latenzmuster im Unterschied zu M/T Zellen eine geringere Vorhersagekraft für die Geruchsstoffkonzentration besitzen als Feuerratenmuster. Außerdem wiesen Ensemble-Feuerraten einen größeren dynamischen Bereich bezüglich der Geruchsstoffkonzentration auf als Latenzen. Durch eine Kombination von schneller (1,25 kHz) [Ca2+] -Bildgebung und whole-cell Patch-Clamp Technik in einzelnen ORNs wurde die zeitliche Entwicklung der dreidimensionalen intrazellulären Ca2+ -Konzentration während eines Depolarisationspulses gemessen. Mit Hilfe von pixelweiser Angleichung eines numerischen Modells wurden Ballungen spannungsabhängiger Ca2+ Kanäle (VGCC) auf der Oberfläche von ORN-Somata lokalisiert. Da der durchschnittliche gemessene VGCC-Kalziumioneneinstrom einen geringen Beitrag im Vergleich zum Ca2+ Generatorstrom darstellt (<80 pA bzw. geschätzt 900 pA), erklärte sich, warum einzelne Aktionspotentiale nicht mittels [Ca2+] Bildgebung gemessen werden konnten. Bezüglich VGCC-Häufung und möglicher Kolokalisation mit Kaliumkanälen hoher Leitfähigkeit (BK) wurde der Effekt von BK Blocker Iberiotoxin auf ORN-Reizantworten untersucht. In einer Untergruppe aller ORNs wurde eine Verringerung der Antwortamplituden nach Anwendung von Iberiotoxin festgestellt. Aus den gezeigten Ergebnissen wurde geschlossen, dass eine wichtige Funktion von Glomeruli im OB die Konversion von Geruchsinformation zwischen Feuerratenkodierung und Latenzkodierung sein müsse.

Neurogenesis occurs in exclusive regions in the adult nervous system, the subventricular zone and dentate gyrus in the brain, and olfactory epithelium (OE) in the periphery. Cell replacement after death or injury, occurs to varying degrees in neural tissue, and is thought to be dependent upon the biological responses of stem and/or progenitor cells. Despite the progress made to identify adult OE and central nervous system (CNS) progenitors and lineage trace their progeny, our spatial and temporal understanding of embryonic OE neuroglial progenitors has been stalled by the paucity of identifiable genes able to distinguish individual candidate progenitors. In the developing CNS, radial glia serve as both neural progenitors and scaffolding for migrating neuroblasts and are identified by the expression of a select group of antigens, including nestin. Here, I show that the embryonic OE contains a novel radial glial-like progenitor (RGLP) that is not detected in adult OE. RGLPs express the radial glial antigens nestin, GLAST and RC2, but not brain lipid binding protein (BLBP), which, distinct from CNS radial glia, is instead found in olfactory ensheathing cells, a result confirmed using lineage tracing with BLBP-cre mice. Nestin-cre-mediated lineage tracing with three different reporters reveals that only a subpopulation of nestin-expressing RGLPs activate the “CNS-specific” nestin regulatory elements, and produce spatially restricted neurons in the OE and vomeronasal organ. The dorsal-medial restriction of transgene-activating cells is also seen in the embryonic OE of Nestin-GFP transgenic mice, where GFP is found in a subpopulation of GFP+ Mash1+ neuronal progenitors, despite the fact that endogenous nestin expression is found in RGLPs throughout the OE. In vitro, embryonic OE progenitors produce three biologically distinct colony subtypes, that when generated from Nestin-cre/ZEG mice, produce GFP+ neurons, recapitulating their in vivo phenotype, and are enriched for the most neurogenic colony subtype. Neurogenesis in vitro is driven by the proliferation of nestin+ progenitors in response to FGF2. I thus provide evidence for a novel neurogenic precursor, the RGLP of the OE, that can be regulated by FGF2, and provide the first evidence for intrinsic differences in the origin and spatiotemporal potential of distinct progenitors during OE development.

Terrestrial salamanders of the genus Plethodon are among many vertebrates possessing both main olfactory and vomeronasal systems, which the Volatility Theory posits are for detection of volatile and soluble olfactory cues, respectively. Further recent work showing a high amount of convergence between the two olfactory subsystems at the level of the central nervous system suggests complementary or overlapping roles for them. This study examined the use of the olfactory subsystems in prey detection from the perspectives of behaviour and neurobiology. Red-backed salamanders, Plethodon cinereus, were observed in standardized behavioural assays with both volatile and soluble prey olfactory cues. Naïve salamanders showed an increase in nosetapping as well as a side preference in the presence of soluble and volatile prey cues when tested in a 22°C day/20°C night room. In a 15°C day /12°C night room, salamanders increased nosetapping in the presence of soluble prey cues. Salamanders showed a pattern of responses that differed based on their previous experience with the assay, as well as the temperature of the testing room. Attempts to study the neurobiology of olfactory function in Plethodon shermani were inconclusive up to this point, but future directions are discussed. This study shows the importance of olfaction in prey detection by salamanders and that prey searching behaviour is exhibited in the exclusive presence of olfactory cues.

Our senses are our mind's window to the outside world and determine how we perceive our environment.Sensory systems are complex multi-level systems that have to solve a multitude of tasks that allow us to understand our surroundings.However, questions on various levels and scales remain to be answered ranging from low-level neural responses to behavioral functions on the highest level.Modeling can connect different scales and contribute towards tackling these questions by giving insights into perceptual processes and interactions between processing stages.In this thesis, numerical simulations of spiking neural networks are used to deal with two essential functions that sensory systems have to solve: pattern recognition and prediction.The focus of this thesis lies on the question as to how neural network connectivity can be used in order to achieve these crucial functions.The guiding ideas of the models presented here are grounded in the probabilistic interpretation of neural signals, Hebbian learning principles and connectionist ideas.The main results are divided into four parts.The first part deals with the problem of pattern recognition in a multi-layer network inspired by the early mammalian olfactory system with biophysically detailed neural components.Learning based on Hebbian-Bayesian principles is used to organize the connectivity between and within areas and is demonstrated in behaviorally relevant tasks.Besides recognition of artificial odor patterns, phenomena like concentration invariance, noise robustness, pattern completion and pattern rivalry are investigated.It is demonstrated that learned recurrent cortical connections play a crucial role in achieving pattern recognition and completion.The second part is concerned with the prediction of moving stimuli in the visual system.The problem of motion-extrapolation is studied using different recurrent connectivity patterns.The main result shows that connectivity patterns taking the tuning properties of cells into account can be advantageous for solving the motion-extrapolation problem.The third part focuses on the predictive or anticipatory response to an approaching stimulus.Inspired by experimental observations, particle filtering and spiking neural network frameworks are used to address the question as to how stimulus information is transported within a motion sensitive network.In particular, the question if speed information is required to build up a trajectory dependent anticipatory response is studied by comparing different network connectivities.Our results suggest that in order to achieve a dependency of the anticipatory response to the trajectory length, a connectivity that uses both position and speed information seems necessary.The fourth part combines the self-organization ideas from the first part with motion perception as studied in the second and third parts.There, the learning principles used in the olfactory system model are applied to the problem of motion anticipation in visual perception.Similarly to the third part, different connectivities are studied with respect to their contribution to anticipate an approaching stimulus.The contribution of this thesis lies in the development and simulation of large-scale computational models of spiking neural networks solving prediction and pattern recognition tasks in biophysically plausible frameworks. / <p>QC 20150504</p>

The mammalian genome encodes five collapsin response mediator protein (CRMP) isoforms. Cell culture studies have shown that the CRMPs mediate growth cone dynamics and neuron polarity through associations with a variety of signal transduction components and cytoskeletal elements. CRMP is also a member of a protein family including the presumably ancestral dihydropyrimidinase (DHP) protein that catalyzes the second step in pyrimidine degradation. In Drosophila, CRMP and DHP proteins are produced by alternatively spliced transcripts of the CRMP gene. The alternative protein forms have a 91% sequence identity, but unique expression patterns. CRMP is found exclusively in neuronal tissues and DHP is ubiquitously expressed in non-neuronal tissues. Comparative analysis of CRMP homologous sequences from insect taxa show CRMP alternative splicing is a common feature and probably represents the ancestral state of this gene family. To investigate the regulatory role of CRMP, loss-of-function mutations of CRMP that lack both proteins were isolated; homozygous animals display DHP-null phenotypes but exhibit no overt developmental or neurological defects. To determine possible interactions of Drosophila CRMP with signaling pathways in which mammalian CRMP has been shown to act, the UAS-GAL4 system was utilized. Phenotypes produced by misexpression of a variety of UAS signal transduction mediator responders were modified in a CRMP mutant background. The modification entails enhancement or suppression of a specific phenotype in a direction that corresponds to the hypothesized involvement of mammalian CRMP in signaling pathways that regulate growth cone dynamics. These data suggest that Drosophila CRMP has a role in cell signaling pathways similar to the role of the mammalian CRMPs. Furthermore, recent findings demonstrate that CRMP plays an important role in learning and memory of mice, leading to the assessment of new phenotypes in the Drosophila CRMP mutants. Tests utilizing the Pavlovian olfactory conditioning assay reveal that loss of CRMP function leads to significant learning, 3 hour memory, and long term memory deficits. Preliminary data also suggest that Drosophila CRMP may be required for normal circadian locomotor rhythms. Collectively, the data presented here demonstrate CRMP’s role in adult behavioral processes and regulating signaling events comparable to mammalian CRMP signaling.

Olfactory sensory neurons (OSNs) undergo continual degeneration and replacement throughout life, a cycle that can be synchronized experimentally by performing olfactory bulbectomy (OBX). OBX induces apoptosis of mature OSNs, which is followed by an increase in the proliferation of progenitor basal cells. Macrophages, functionally diverse immune effector cells, phagocytose the apoptotic OSNs and regulate the proliferation of basal cells. This provides an advantageous environment to study how macrophages regulate neuronal death, proliferation, and replacement. The purpose of this dissertation was to identify the cellular and molecular mechanisms by which macrophages regulate the degeneration/proliferation cycle of OSNs. Macrophages were selectively depleted using liposome-encapsulated clodronate (Lip-C). Intranasal and intravenous administration of Lip-C decreased the number of macrophages in the OE of sham and OBX mice by 38% and 35%, respectively, compared to mice treated with empty liposomes (Lip-O). Macrophage depletion significantly decreased OE thickness (22% and 21%, p<0.05), the number of mature OSNs (1.2- and 1.9-fold, p<0.05), and basal cell proliferation (7.6- and 3.8-fold, p<0.05) in sham and OBX mice, respectively, compared to Lip-O mice. Additionally, at 48 h following OBX, OSN apoptosis increased significantly (p<0.05) in the OE of Lip-C mice compared to Lip-O mice. A microarray analysis was performed to identify the genomic changes underlying the cellular changes associated with macrophage depletion. There were 4,024 genes with either a significant interaction between group (Lip-C vs. Lip-O) and treatment (OBX vs. sham) or a significant main effect. There were a number of significantly regulated immune response and cytoskeletal genes, and genes encoding neurogenesis regulators and growth factors, most of which were expressed at lower levels in Lip-C mice compared to Lip-O mice. Sdf1, the ligand for the chemokine receptor Cxcr4 involved in leukocyte trafficking, axon guidance, and cell migration, was localized to macrophages on the protein level. Additionally, the microarray expression pattern of Hdgf, a growth factor that promotes neuronal survival and proliferation, was validated on the protein level using immunohistochemistry. HDGF appeared to be localized to basal cells and OSNs where it could act as a proliferative or survival factor whose expression is regulated in part by macrophages.