Im Gegensatz zum peripheren Nervensystem (PNS) ist die Regenerationsfähigkeit im adulten zentralen Nervensystem (ZNS) von Vertebraten sehr eingeschränkt. Diese eingeschrängte Regenerationsfähigkeit wird im Wesentlichen durch das Vorhandensein von Myelin im adulten ZNS determiniert. Einerseits ist dieses Lipid für die Stabilisierung und Ernährung von Axonen sowie für die schnelle Reizweiterleitung unbedingt notwendig, andererseits stellt es den größten Inhibitor axonaler Regeneration dar. Myelin ist außerdem Zielstruktur diverser ZNS Pathologien, wie z.B. der Multiplen Sklerose. Für das Verständnis dieser Pathologien sowie der auswachsinhibitorischen Wirkung von Myelin wurde der Hippocampus als eine der plastischten ZNS Regionen gewählt. Dazu waren genaue Kenntnisse der Myeloarchitektur dieses Gebietes notwendig. Nach Etablierung einer zuverlässigen Detektierung für Myelin konnten in der vorliegenden Arbeit detailliert Myelinisierungsvorgänge im sich entwickelnden, im adulten und im deafferenzierten Hippocampus der Ratte analysiert werden. Während der Entwicklung erreichen die ersten entorhinale Axone den Hippocampus bereits am embryonal Tag 17 (E17); Myelin kann jedoch erst am postnatal Tag 17 (P17) lichtmikrokopisch nachgewiesen werden. Die Anzahl myelinisierter Fasern erreicht um den P25 ein Verteilungsmuster, welches dem von adulten Tieren gleicht. Nach Entorhinaler Cortex Läsion (ECL), bei der die Durchtrennung des Tractus perforans (PP) eine Denervation des Hippocampus bewirkt, kommt es zu einem langanhaltenden Verlust von Myelin. Zehn Tage nach Läsion (10 dal), also zum Zeitpunkt maximaler Aussprossung (Sprouting), kommt es zu einem Wiederkehren myelinisierter Fasern. Mehrere myelin-assoziierte Proteine, mit wachstumshemmenden Eigenschaften sind bekannt, wie z.B. die Familie der Nogo Gene (Nogo; englisch, kein Durchkommen). Diese werden ganz entschieden für den Verlust der Regenerationsfähigkeit des adulten ZNS verantwortlich gemacht. In der vorliegenden Arbeit wird die Expression der drei Nogo Gene (Nogo-A, -B, - C) und deren Rezeptor (Ng66R) während der postnatalen Entwicklung, im adulten ZNS sowie nach Läsion beschrieben. Ein erster überraschender Befund war die neuronale Expression der Nogos, die bisher nur in Oligodendrocyten nachgewiesen worden war. Zu einem Zeitpunkt, an dem entorhinale Fasern bereits in den Hippocampus eingewachsen, aber noch nicht myelinisiert sind (P0), wird Nogo-A, -B und Ng66R mRNA mit Ausnahme der Körnerzellschicht des Gyrus dentatus in allen Zellschichten des sich entwickelnden Hippocampus detektiert. Nogo-C und myelin basic protein (MBP) mRNA, werden erst am P15 expremiert, zu einem Zeitpunkt also, an dem myelinisierte Fasern erstmalig im Hippocampus auftreten. MBP wird ausschließlich in glialen, Nogo-C hingegen hauptsächlich in neuronalen Zellen exprimiert. Nach Deafferenzierung zeigt sich eine dynamische und Isoform- spezifische Regulation aller Nogo Transkripte. So zeigen die als erste von der Deafferenzierung betroffenen Körnerzellen zu Beginn der Waller`schen Degeneration sowie der neuronalen und glialen Antwort, eine starke Erhöhung aller Nogo Transkripte. Zum Zeitpunkt der maximalen Aussprossung kam es zu einem signifikanten Abfall der Nogo-C und Ng66R mRNA Expression, währendessen Nogo-A und Nogo-B bereits wieder das Kontrollniveau erreicht hatten. Vor allem im contralateralen Hippocampus, dem Hauptquellgebiet sproutender Fasern, imponierte die Runterregulation von Ng66R mRNA und zeigte erst nach Abschluß von axonalen Sproutingprozessen und der Synapsenformation wieder vergleichbare Werte mit den Kontrolltieren. Diese Korrelation der erniedrigten Ng66R Expression im contralateralen Hippocampus und dem axonalen Einwachsen in den deafferenzierten Hippocampus, läßt eine reduzierte axonale Ansprechbarkeit auf den Neuriten-Auswachshemmer Nogo-A vermuten, da bekannt ist, dass Axone, die kein Ng66R exprimieren, nicht durch die Nogo Gene im Wachstum gehemmt werden. Zusammenfassend kommt es während der Entwicklung und in der Reorganisationsphase zu einer spezifischen und geordneten Myelinisierung im Hippocampus. Die neuronale Expression von Nogo- A, -B und -C in einer so plastischen ZNS- Region unterstützt die Hypothese, dass den Nogo- Genen neben der reinen Hemmung von axonalen Auswachsen weitere Funktionen zuzuordnen sind. So scheinen sie vor allem während der Entwicklung und während der Stabilisierungsphase der hippocampalen Reorganisation eine wichtige Rolle einzunehmen. Die hier dargestellten Daten zeigen auf, dass vor einem therapeutischen Einsatz von Nogo- Antagonisten nach Schädigung deren Verträglichkeit bzw. unerwünschte Nebeneffekte ausgeschlossen werden müssen. / Compared to the peripheral neuronal system (PNS) the reorganisation capacity in the adult central neuronal system (CNS) is highly restricted. One important reason for the lack of reorganisation is the existence of myelin in the CNS. Myelin is crucial for the stabilization of axonal projections in the developing and adult mammalian brain. However, myelin components also act as a non-permissive and repellent substrate of outgrowing axons. In these thesis the appearance of mature, fully myelinated axons during hippocampal development and following entorhinal cortex lesion with the myelin-specific marker Black Gold is reported. Althrough entorhinal axons enter the hippocampal formation at the embryonic day 17, light and ultrastructural analysis revealed that mature myelinated fibres in the hippocampus occur in the second postnatal week. During postnatal development, increasing numbers of myelinated fibers appear and the distribution of myelinated fibers at postnatal day 25 was similar to that found in the adult. After entorhinal cortex lesion, a specific anterograde denervation in the hippocampus takes place, accompanied by a long- lasting loss of myelin. Quantitative analysis of myelin and myelin breakdown products at different time points after lesion revealed a temporally close correlation to the degeneration and reorganisation phases in the hippocampus. In conclusion, it could be shown that the appearance of mature axons in the hippocampus is temporally regulated during development. Reappearing mature axons were found in the hippocampus following axonal sprouting. Various myelin-associated proteins, with neurite inhibition properties are known. One is the family of Nogo genes (no go). They are distinctly responsible for the lack of reorganisation. In these thesis the expression pattern of Nogo-A, Nogo-B, Nogo-C and Nogo-66 receptor (Ng66R) mRNA during hippocampal development and lesion induced axonal sprouting is reported. The first surprising result was the neuronal expression of all Nogos, who were supposed to be only expressed by oligodendrocytes. Nogo-A, Nogo-B and Ng66R transcrips preceded the process of myelination and were highly expressed at postnatal day zero (P0) in all principal hippocampal cell layers, with the exception of dentate granule cells. Only a slight Nogo-C expression was found at P0 in the principal cell layers of the hippocampus. During adulthood, all Nogo splice variants and their receptor were expressed in the neuronal cell layers of the hippocampus, in contrast to the myelin basic protein mRNA expression pattern, which revealed a neuronal source of Nogo gene expression in addition to oligodendrocytes. After hippocampal denervation, the Nogo genes showed an isoform-specific temporal regulation. All Nogo genes were strongly regulated in the hippocampal cell layers, wheras the Ng66R transcrips showed a significant increase in the contralateral cortex. These data could be confirmed on protein levels. Futhermore, Nogo-A expression was up-regulated after kainat- induced seizure. These data show that neurons express Nogo genes with a clearly distinguishable pattern during development. This expression is further dynamically and isoform-specifically altered after lesioning during the early phase of structural rearrangements. Thus, these results indicate a role for Nogo-A, -B and –C during development and during stabilisation phase of hippocampal reorganization. Taken together with these data, the findings that neurons in a highly plastic brain region express Nogo genes supports the hypothesis that Nogo may function beyond its known neuronal growth inhibition activity in shaping neuronal circuits.

Woody plant encroachment is one of the major problems worldwide because it affects negatively the herbaceous layer, which provide forage for livestock production. However, the role of ruminants particularly browsers in the dispersal of woody plant seeds still remains a concern for farmers interested in grass production. Seedpods of various woody plant species constitute a crucial part of the diet of herbivores during the dry season because of their high nutritional quality compared to herbaceous material. The interaction of associated diet quality, seed characteristics and animal species among other factors play a pivotal part in the success of livestock faecal seeds dispersion. Furthermore, dispersed seeds that successfully grow into mature woody plants become an important source of protein for herbivores. The use of woody plants as a source forage is known to be limited by plant secondary metabolites (PSMs) such as condensed tannins. The objectives of this study were to determine 1) the effects of condensed tannins and crude protein of Vachellia tortilis and Dichrostachys cinerea pods in seed recovery and germination fed to goats, and 2) the effects of diet mixing on the feed intake of plant species by goats. In the first experiment, a total of 12 female indigenous goats and 12 female Pedi sheep were utilised in this study, with the average body weights of 29.50 kg ± 1.60 (S.E) and 28.70 kg ± 1.60, respectively. Twelve goats were grouped into two groups of six goats per group, one group was fed D. cinerea pods and the other group was fed V. tortilis pods. The group of 12 sheep were divided similarly, the one group was fed D. cinerea pods and the other group was fed V. tortilis pods. Each animal was given V. tortilis and D. cinerea pods at 2.50% of their body weight. All animals were allowed to consume D. cinerea or V. tortilis pods within 24 h, after which the remaining pods were collected and weighed. Faecal collection commenced immediately after the 24 h pods feeding and was carried on until no seeds were discovered in faeces. All faeces extracted from sheep and goats were collected daily in the morning from the faecal bags. In the second experiment, a total of 24 indigenous goats with average body weight of 26.6 kg ± 0.51 were utilised. Goats were arbitrarily selected and grouped into four groups of six goats per group (goats were placed individually in 2 m2 pens). Each group was fed one of the following diets: diet one - Searsia lancea, diet two - S. pyroides, diet three - Euclea crispa and diet four - was a combination of the three plant species (Searsia lancea, S. pyroides and Euclea crispa). Searsia lancea, S. pyroides and E. crispa branches were collected every morning prior to feeding, and were weighed before offering the animals. Refusals were gathered and weighed, and intake was calculated as distinction between weight in and refusals. Plant species foliage were analysed for crude protein, condensed tannin, acid detergent lignin, acid detergent fibre and neutral detergent fibre. During the first experiment, the cumulative percentage seed recovery of V. tortilis from goats (46.00 % ± 1.90) and sheep (52.00 % ± 2.93) was significantly higher than D. cinerea from goats (13 % ± 1.47) and sheep (24.00 % ± 1.16). Germination percentage of D. cinerea seeds that passed through the gastro-intestinal tract of goats (33.12 % ± 2.94) and sheep (36.00 % ± 2.68) was significantly higher than V. tortilis seeds that passed through the gastro-intestinal tract of goats (28.98 % ± 2.68) and sheep (23.04 % ± 2.81). Average D. cinerea (34.56 % ± 1.99) and V. tortilis (26.02 %± 2.10) seeds that went through the gastro-intestinal of goats and sheep had a significantly higher germination rate than the control (i.e. no passage through the gut; D. cinerea = 2.31 % ± 1.55, V. tortilis = 5.07 % ± 2.68). The high mean cumulative percentage seed recovery of V. tortilis (18.80 %) may be attributed to the relatively higher crude protein than D. cinerea (12.20 %). This may encourage animal seed dispersal and germination of woody plant species with relatively high crude protein content. In the second experiment, Searsia lancea contained 8.50 % CP, 21.46 % acid detergent fibre (ADF), 12.50 % ADL and 39.37 % NDF. Searsia pyroides had 9.03 % CP, 27.07 % ADF, 10.89 % ADL and 40.30 % NDF. Euclea crispa had 6.19 % CP, 26.20 % ADF, 16.63 % ADL and 30.02 % NDF. Mixed diet (combination of the three plant species) had 8.96 % CP, 23.72 % ADF, 11.13 % ADL and 38.28 % NDF. Searsia lancea had 2.70 % of CTs while S. pyroides had 5.20 % CT, E. crispa had 6.44 % CT and mixed diet had 7.20 % CT. The mean dry matter intake varied significantly among dietary groups (P < 0.001). Similarly, goats offered a mixed diet consumed more CTs (P < 0.01) than those offered individual forage species. The high mean cumulative percentage seed recovery of V. tortilis may be attributed to the higher crude protein of V. tortilis (18.80 %) than D. cinerea (12.20 %). Higher passage rate may encourage animal seed dispersal and germination of plant species. The results from experiment two support the postulation that animals foraging in mixed diet systems consume more PSMs and achieve higher dry matter intake than animals confined to monocultures or single species feeding systems. Given that woody plant encroachment is already reducing farm-grazing capacities in African savannas and this problem is predicted to double by 2050, strategies that improve herbivore ability to consume woody plants will increase forage availability and inform bush control programmes and policies. Moreover, the concomitant increase in CTs by goats exposed to diets with diverse species also has positive implications for animal / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

Wetland plant species rely largely on vegetative reproduction. Although all types of clonal growth organs are found in wetlands, special adaptations, e.g., turions, fragmentation and budding, are more frequent in true aquatic communities. However, root-sprouting is underrepresented, although it can be beneficial under disturbed conditions. This thesis focuses on ecological constrains potentially hindering root-sprouting in wetlands. This ability was studied in a wetland herb Rorippa palustris in relation to life history, injury timing and carbon economy of a plant. In addition, plant regeneration following submergence and severe disturbance was assessed.

No description available.

Les neurones dopaminergiques de la substance noire (SNc) sont les plus vulnérables à la dégénérescence dans la maladie de Parkinson et ses modèles animaux. Suite à des travaux antérieurs et à des résultats préliminaires du laboratoire Trudeau, notre hypothèse actuelle suggère que la très grande taille de l'arborisation axonale des neurones de la SNc soit un facteur clé à l'origine de leur vulnérabilité, puisque cet état devrait être associé à un taux élevé de phosphorylation oxydative et de production de radicaux libres. En accord avec cette hypothèse, les autres populations dopaminergiques, dotées d'arborisations de moindre taille, résistent mieux aux lésions expérimentales et à la maladie chez l'humain. L'objectif du présent projet était de développer un modèle murin dans lequel les neurones de la SNc présentent une taille d'arborisation axonale plus grande, se rapprochant davantage de celle observée chez l'humain et en reproduisant la vulnérabilité, ce qui pourrait représenter une percée importante dans l'identification de nouvelles approches thérapeutiques. Basée sur le bourgeonnement axonal compensatoire des neurones dopaminergiques suite à des lésions partielles, la méthode utilisée fut l'injection unilatérale intranigrale de la toxine 6-hydroxydopamine (6-OHDA) à quelques jours de vie (P5), en visant l'élimination de 50% des neurones de la SNc. Un immunomarquage contre la tyrosine hydroxylase (TH), enzyme de synthèse de la dopamine, ainsi qu'une quantification du signal TH dans le striatum et des comptes neuronaux stéréologiques ont permis de quantifier la lésion partielle et de mettre en évidence la présence d'une croissance axonale compensatoire des neurones dopaminergiques survivants, à 10 et 90 jours post-lésion, suggérant une compensation précoce. Afin de mettre en évidence l'origine du bourgeonnement axonal, nous avons injecté un vecteur viral de type AAV encodant une protéine fluorescente (EYFP) dans la SNc ou la VTA des animaux adultes. Nos résultats confirment la présence de neurones nigrostriés à plus grande arborisation suivant une lésion unilatérale précoce à la 6-OHDA, dont la vulnérabilité accrue pourra être évaluée dans des expériences à venir par des protocoles lésionnels au MPTP, une toxine permettant de modéliser la maladie de Parkinson chez la souris. / Dopaminergic neurons of the substantia nigra (SNc) are amongst the most vulnerable to neurodegeneration in Parkinson's disease and its animal models. According to previous work and preliminary results in our laboratory, our present hypothesis postulates that the large axonal arborisation size of SNc neurons is a key driving factor in their vulnerability, since this characteristic is associated with increased oxidative phosphorylation levels and free radicals production. In agreement with this hypothesis, other dopaminergic populations with smaller axonal arbors better resist to experimental lesions and to the disease process in humans. The current project aims to develop a mouse model in which SNc neurons present an axonal arborisation of increased size, closer to what is encountered in humans, thus reproducing their vulnerability, which could represent an important breakthrough in the identification of new therapeutic approaches. Based on compensatory axonal sprouting of dopaminergic neurons following partial lesions, the method used was the unilateral intranigral injection of the toxin 6-hydroxydopamine (6-OHDA) at an early age (P5), to induce the loss of approximately 50% of SNc neurons. Immunostaining against tyrosine hydroxylase (TH), an enzyme required for the synthesis of dopamine, TH signal quantification in the striatum and stereological counting of neurons allowed for the quantification of the partial lesion and demonstrated compensatory axonal sprouting at 10 and 90 days post-lesion, with our results suggesting an early compensation. To better characterize the origin of axonal sprouting, we injected an AAV viral vector encoding a fluorescent protein (EYFP) in either the SNc or the VTA of adult animals. Our results confirm the presence of nigrostriatal neurons with increased arborisation sizes following early unilateral lesion using 6-OHDA, whose increased vulnerability will be evaluated in future experiments through lesion protocols using MPTP, a toxin used to model Parkinson's disease in mice.

No description available.

Disturbance is one of most important selective factor causing removal of plant biomass. Man-made habitats are characterized by strong and unpredictable disturbances, providing bare soil surface colonized by plants with short life cycle. Populations of the short-lived plants are, however, vulnerable to the strong disturbance removing all stem parts with reserve axillary meristems in the case it occurs before plant flowering and fruiting. Nevertheless, 2 % of annual and 14 % of biennial plants are able to overcome meristem limitation by adventititous sprouting from hypocotyle or/and roots. This thesis is composed of four original studies describing the occurrence of adventitious sprouting in natural populations of 22 monocarpic weeds of Central Europe and one species in Indonesia. The studied phenomenon was analyzed in relation to various environmental factors and plant traits in the field and in experimental conditions.

Indiana University-Purdue University Indianapolis (IUPUI) / Posttraumatic epilepsy, the development of temporal lobe epilepsy (TLE) following traumatic brain injury, accounts for 20% of symptomatic epilepsy. Reorganization of mossy fibers within the hippocampus is a common pathological finding of TLE. Normal mossy fibers project into the CA3 region of the hippocampus where they form synapses with pyramidal cells. During TLE, mossy fibers are observed to innervate the inner molecular layer where they synapse onto the dendrites of other dentate granule cells, leading to the formation of recurrent excitatory circuits. To date, the molecular mechanisms contributing to mossy fiber sprouting are relatively unknown. Recent focus has centered on the involvement of tropomycin-related kinase receptor B (TrkB), which culminates in glycogen synthase kinase 3β (GSK3β) inactivation. As the neurite outgrowth promoting collapsin response mediator protein 2 (CRMP2) is rendered inactive by GSK3β phosphorylation, events leading to inactivation of GSK3β should therefore increase CRMP2 activity. To determine the involvement of CRMP2 in mossy fiber sprouting, I developed a novel tool ((S)-LCM) for selectively targeting the ability of CRMP2 to enhance tubulin polymerization. Using (S)-LCM, it was demonstrated that increased neurite outgrowth following GSK3β inactivation is CRMP2 dependent. Importantly, TBI led to a decrease in GSK3β-phosphorylated CRMP2 within 24 hours which was secondary to the inactivation of GSK3β. The loss of GSK3β-phosphorylated CRMP2 was maintained even at 4 weeks post-injury, despite the transience of GSK3β-inactivation. Based on previous work, it was hypothesized that activity-dependent mechanisms may be responsible for the sustained loss of CRMP2 phosphorylation. Activity-dependent regulation of GSK3β-phosphorylated CRMP2 levels was observed that was attributed to a loss of priming by cyclin dependent kinase 5 (CDK5), which is required for subsequent phosphorylation by GSK3β. It was confirmed that the loss of GSK3β-phosphorylated CRMP2 at 4 weeks post-injury was likely due to decreased phosphorylation by CDK5. As TBI resulted in a sustained increase in CRMP2 activity, I attempted to prevent mossy fiber sprouting by targeting CRMP2 in vivo following TBI. While (S)-LCM treatment dramatically reduced mossy fiber sprouting following TBI, it did not differ significantly from vehicle-treated animals. Therefore, the necessity of CRMP2 in mossy fiber sprouting following TBI remains unknown.