In this study the spawning migration activity, habitat choice (depth and temperature), foraging activity, duration of stay in the spawning site and homing-behavior was studied for anadromous pike (Esox lucius) in the wetland Österby myr, Gotland. The spawning migration was concentrated to dusk and dawn, a homing behavior was confirmed and there was no significant difference between sexes with respect to duration of stay. A difference in habitat choice and small variation in foraging activity between individuals was established.

Stem cells hold great promise for treating various degenerative diseases. However, the outcomes of preclinical and clinical cell therapy studies are still not close to our expectation. The unsatisfactory outcomes of cell therapy are at least partially due to: 1) insufficient homing of implanted stem cells into target organs and 2) use of heterogeneous cell populations for cell therapy. Therefore, there is a need to develop effective guiding technique for stem cells to migrate to the target organs and to isolate effective stem cell populations. In this project, I developed a microfluidics-based method for selecting chemotactic adipose-derived stem cells (ASCs) to epidermal growth factor (EGF). This method integrates cell patterning, chemotaxis and cell extraction on a single microfluidic device. Post-extraction analysis confirmed the higher chemotactic ability of the extracted cells to EGF. The extracted chemotactic ASCs shows up-regulated surface expression of EGF receptor and its downstream signaling event upon EGF stimulation.

In the following study, I tried to link hormonal background conditions to successful olfactory imprinting in sockeye salmon by employing behavioural, endocrinological and electrophysio logical experiments. In the initial experiments, sockeye salmon were exposed to potential imprinting odorants, with or without additional treatment with thyroid hormones, during several juvenile stages between fertilization and beyond the PST. After two years of rearing, these fish were tested for behavioural responses to test odorants in two behavioural arenas. Neither immature nor mature fish reacted behaviourally to the odorants that they had been exposed to previously. Therefore, exposure of juveniles to odorants did not lead to imprinting to those odorants under hatchery rearing conditions. In contrast, juvenile fish that were exposed to test odorants and treated with a combination of T₃ and T₄ (in all cases) or T₃ (in one case) the two most common forms of thyroid hormones, did exhibit an odorant recognition response two years later. However, the response differed between immature and mature fish. Mature fish were attracted to the imprinting odorant, whereas immature fish were repelled by the it. When immature fish were injected with GnRH before testing, their behavioural response was reversed. No behavioural response could be detected in fish that had been challenged with either T₃ or T₄ alone, in contrast to a combined treatment with both forms. Thus, I found evidence that a combination of T₃ and T₄ initiated imprinting and that GnRH motivated odorant recognition. To examine the underlying hormonal processes, I first determined plasma thyroid hormone concentrations in sockeye salmon before and after hormonal challenges with thyroid hormones or GnRH. In addition, the activity of the deiodinase enzyme that converts T₄ into the other possible forms of thyroid hormones was investigated in sensory and non-sensory tissues. The results suggested that only a combined T₃T₄ treatment increased the availability of both thyroid hormone forms in blood plasma, while a separate challenge with T₄ suppressed T₃ availability and vice versa. Moreover, the results provided evidence for deiodinase activity in the olfactory epithelium and the retina and demonstrated that GnRH can modulate the T₄ conversion process. This inform ation was helpful for planning and interpretation of the remaining experiments. Results obtained from a classical conditioning paradigm (heart-rate-conditioning), provided support for the hypothesis that GnRH lowers the threshold to an imprinting odorant and that the influence of GnRH was not restricted to an enhancement of motivation. To investigate whether hormonal action could also modulate the sensitivity of the peripheral olfactory system, electrophysiological responses from the olfactory epithelium (electro-olfacto-grams or EOGs) were recorded. The EOG results established that thyroid hormones and GnRH increased the EOG response of adult naïve (never imprinted to an odorant) fish, as did maturity. In the last experiments, I conducted EOG recordings on fish that had been imprinted at a juvenile stage. In summary, EOG recordings revealed that the imprinting process increased sensitivity to the imprinting odorant at maturity, while sensitivity in immature fish was decreased in comparison to non-imprinted fish. In combination with my behavioural results, this could explain why salmon do not enter their natal stream before they reach maturity. At maturity however, I also encountered desensitization to non-imprinting odorants, which might increase the ability to focus the olfactory system to the task of homing. / Graduate

The purpose of the study is to find out how Oligocottus maculosus homes and what mechanisms or sensory channels are involved in the homing process. The species' home range was defined from pool fidelity and movement observations. Evidence was obtained to show that the fish homes when displaced to unfamiliar areas. The roles of season, distance and direction of displacement as well as sex, size and experience of the fish in homing were assessed. The roles of both vision and olfaction and the related environmental cues were also investigated. Some fish showed strict fidelity to specific tide-pools while others moved from one pool to another but limited themselves to a restricted group of neighbouring pools. The range of movement seldom exceeded an area of about one hundred square feet which was defined as the maximum home range. Homing, i.e. striving to return to its home range instead of going to other equally habitable areas, was observed throughout the year; homing was most successful between March and August. This seasonal variation was probably the result of mortality owing to the seasonal sea conditions. In my study O. maculosus had been shown to home from as far as one thousand feet. A decrease in homing returns with increasing distances from home was observed. This was attributed to the effects of wave action which decreased the chances of survival with increasing distances from home. Direction of displacement did not seem to affect homing performance and no difference in homing performance was observed between the sexes or among fish of different sizes. Fish which were shown to home at least once seemed to home better than naive fish though no further improvement in homing was observed when the former were repeatedly displaced. There is a strong indication that there is inherent variability in homing ability, that some fish home better than others and that learning per se is not important. Displacement experiments conducted with blind and anosmic fish had shown that blind fish could home better than anosmic fish indicating that olfaction, not vision, is more important in homing. Studies on the ability to home on cloudy nights, the ability to discriminate sea water from different sources in the laboratory and the ability to return to pools with the immediate environment destroyed further indicated the importance of olfaction in the homing mechanism of O. maculosus. How the fish finds its way from the release point to its home range is still not known with certainty. However, it is suggested that the fish homed by (i) following odour streams originating from its home area or (ii) an exploratory search process or (iii) a combination of the two processes. / Science, Faculty of / Zoology, Department of / Graduate

Le plateado ou dorado - Brachyplatystoma rousseauxii (Pimelodidae, Siluriformes) est un grand poisson-chat Amazonien d’intérêt commercial, qui présente un des cycles de vie les plus surprenants et énigmatiques, avec la plus grande migration connue en eaux douces, entre l’estuaire de l'Amazone et les têtes de fleuves en Amazonie occidentale. Le but de ce travail était de déterminer, au niveau moléculaire, la position phylogénétique du plateado dans la famille Pimelodidae ainsi que sa structure génétique dans le Haut Madera (Villa Bella–VB, Cachuella Esperanza–CE, Puerto Maldonado–PM, Rurrenabaque–RU, Puerto Villarroel–PV) et ouest de l'Amazonie (Iquitos–IQ) bassins (Bolivie et Pérou). Les relations phylogénétiques ont été définies par une analyse du maximum de vraisemblance (ML) des séquences nucléotidiques de deux gènes mitochondriaux (Région de Contrôle–RC, ~ 900 pb, 32 taxons; Cytochrome Oxydase 1-CO1, ~ 650 pb, 61 taxons), et d’un gène nucléaire (F-reticulon4–RTN4, ~ 1700 pb, 38 taxons). La structure génétique des populations a été évaluée par le polymorphisme de longueur de neuf microsatellites (284 inds) et par les variations de séquence de la RC (461 inds + 45 en provenance du Brésil, disponibles dans GenBank). Les variations de fréquences des microsatellites ont été utilisées pour identifier les unités panmictiques (clusters) les plus probables dans l'ensemble des données, à travers une approche bayésienne (BAPS), après avoir démontré une déviation significative à l'équilibre de Hardy-Weinberg (HWE) quand l’ensemble des données étaient analysé comme faisant partie d’une seule unité.L’analyse phylogénétique concaténée (ML) a montré que la famille Pimelodidae était un groupe monophylétique. Les résultats les plus notables de la phylogénie sont la monophylie peu soutenue (77%) de la tribu Brachyplatystomatini et la non-monophylie des Brachyplatystoma. Seul le sous-genre Malacobagrus (B. rousseauxii + (B. filamentosum + B. capapretum)), défini morphologiquement, s’est avéré monophylétique. Ces résultats suggèrent que le genre Brachyplatystoma pourrait contenir Platynematichthys ou pourrait être limité au sous-genre Malacobagrus.L'analyse des microsatellites sur l'ensemble des échantillons (ouest Amazone + haut Madera) a montré un écart significatif á la panmixia, ainsi que sur l'ensemble des échantillons du haut Madera. A la lumière de ces résultats, l’approche bayésienne a été développée, montrant qu'au moins trois clusters (1, 2, 3) sont présents dans les bassins du haut Madera et de l'ouest de l'Amazone, avec des répartitions qui se chevauchent partiellement. En parallèle á l'identification des clusters, il a été mis en évidence une différence significative au sein de B. rousseauxii entre l’ouest de l'Amazonie et le haut Madera bassin.L'analyse généalogique (ML) des séquences de la RC a montré une topologie en peigne, sans groupe d'haplotypes montrant une histoire commune. En revanche, l'analyse des fréquences haplotypiques a révélé l’existence de 4 haplogroupes, liés à la géographie. Un haplogroupe a été identifié le long de l'axe principal de l’Amazonas-Solimões (Belem-Brésil et Iquitos-Pérou) et 3 autres dans le haut Madera (VB; CE+MD; RU+PV), organisés selon une tendance aval - amont. Ainsi, nous observons d’un coté 3 populations (clusters) avec une distribution géographique partiellement chevauchante, et de l’autre quatre haplogroupes positionnés selon une logique géographique. Le scenario le plus probable implique un comportement de homing des individus du cluster 1 (homing à l’échelle des grands sous-bassins), qui préfèrent ou tendent à retourner dans le sous-bassin du Madera. Les 3 populations coexisteraient alors dans le haut Madera en se reproduisant à des périodes (phénologie) ou à des endroits différents (ségrégation spatiale). Enfin, les résultats sont discutés à la lumière des résultats précédemment publiés dans le bassin de l'Amazone et des menaces qui pèsent sur l'espèce dans le bassin du Madera. / The Plateado or Dorado - Brachyplatystoma rousseauxii (Pimelodidae, Siluriformes) is a commercial migratory catfish species with one of the most surprising and enigmatic life histories in the Amazon basin, involving the largest migration known for a freshwater species, between the estuary and the head waters in the Andean piedmont. The aim of the present work was to determine the molecular phylogenetic position of the Plateado in the Pimelodidae family and its population genetic structure in the Upper Madera (Villa Bella – VB, Cachuella Esperanza – CE, Puerto Maldonado – PM, Rurrenabaque – RU, Puerto Villarroel - PV) and Western Amazon (Iquitos - IQ) basins (Bolivia and Peru). The phylogenetic relationships were defined through a Maximum Likelihood (ML) analysis of nucleotide sequences of two mitochondrial (Control Region – CR, ~ 900 pb, 32 taxa; Cytochrome Oxidase 1 – CO1, ~ 650 bp, 61 taxa), and a nuclear fragment (F-reticulon4 - RTN4, ~1700 bp, 38 taxa). The population genetic structure was evaluated through the length polymorphism of nine microsatellites (284 inds) and CR sequence variations (461 inds + 45 from Brazil available in GenBank). Microsatellites frequencies variations were used to identify through a Bayesian approach (BAPS) the most probable panmictic units (clusters) in the whole data, after previous demonstration of a deviation to Hardy-Weinberg Equilibrium (HWE). The ML phylogenetic concatenated analysis showed the Pimelodidae family as a monophyletic group, with the genera Phractocephalus and Leiarius as basal lineages. The most notable results in the phylogeny were the not well-supported monophyly (77%) of the tribe Brachyplatystomatini and the non-monophyly of Brachyplatystoma. Only the morfologically defined subgenus Malacobagrus (B. rousseauxii + (B. filamentosum + B. capapretum)) was recovered as monophyletic. These results suggest that Brachyplatystoma could contain Platynematichthys or be restricted to the subgenus Malacobagrus, and the other species be related to distinct (earliest) genera, in agreement with another study carried out in parallel with other markers.Microsatellite analysis of the whole data (Western Amazon + Upper Madera) showed a significant departure of the HWE expectations, as well as the analysis of the whole data from the Upper Madera region. In the light of these results, the Bayesian approach has been implemented, showing that at least three clusters (1, 2, 3) are present in the Upper Madera and Western Amazon basins with partial overlapping distribution.To the margin of the cluster identification, it was evident the significant difference between Western Amazon (Iquitos region) and the Upper Madera basin.The genealogical analysis (ML) of the CR sequences showed a generalized comb-like topology without group of haplotypes with common ancestry. On the other hand, CR frequency analysis showed the conformation of four haplogroups associated to geography. One haplogroup was identified along the main axis of the Amazonas-Solimões, from Belem (Brazil) to Iquitos (Peru), and three other haplogroups were observed in the Upper Madera basin (VB; CE+PM; RU+PV), positioned in a downstream - upstream pattern.Hence, we observed on the one hand three genetic populations (clusters), distributed in partially overlapping geographical areas, and on the other hand four haplogroups, positioned according to a geographical pattern. The most probable scenario involves a homing behavior of individuals from cluster 1 (homing at the scale of large watersheds), which prefer or tend to return to the Madera basin, with the three populations coexisting within the upper Madera because they reproduce at different moments (phenology) or different places (spatial segregation). Finally, the results are discussed in the light of previous results in the Amazon basin and the threats to the species in the Madera basin (p.e. fragmentation by dams, overfishing, climate variability, among other).

Topical antigen (Ag) application mimics natural Ag exposure across the skin. Soluble Ag introduced through this route requires cross-presentation by dendritic cells (DCs) to generate CD8 T cell responses, including skin-homing T cells. DCs process Ag for display to other immune cells, and stimulate T cells to release cytokines or directly lyse infected cells. Some T cells are further stimulated to express homing markers allowing them to enter non-lymphoid tissue such as the skin or the gut.

International Telemetering Conference Proceedings / October 18-21, 2004 / Town & Country Resort, San Diego, California / This paper presents an alternative approach to high-precision bearing estimation for navigation and guidance in homing and docking of underwater vehicles. This new technique is significantly simpler than the conventional methods in terms of computation complexity and yet produces results of superior precision and consistency.

<p>Homing endonucleases are rare-cutting enzymes that cleave DNA at a site near their own location, preferentially in alleles lacking the homing endonuclease gene (HEG). By cleaving HEG-less alleles the homing endonuclease can mediate the transfer of its own gene to the cleaved site via a process called homing, involving double strand break repair. Via homing, HEGs are efficiently transferred into new genomes when horizontal exchange of DNA occurs between organisms.</p><p>Group I introns are intervening sequences that can catalyse their own excision from the unprocessed transcript without the need of any proteins. They are widespread, occurring both in eukaryotes and prokaryotes and in their viruses. Many group I introns encode a HEG within them that confers mobility also to the intron and mediates the combined transfer of the intron/HEG to intronless alleles via homing.</p><p>Bacteriophage T4 contains three such group I introns and at least 12 freestanding HEGs in its genome. The majority of phages besides T4 do not contain any introns, and freestanding HEGs are also scarcely represented among other phages.</p><p>In the first paper we looked into why group I introns are so rare in phages related to T4 in spite of the fact that they can spread between phages via homing. We have identified the first phage besides T4 that contains all three T-even introns and also shown that homing of at least one of the introns has occurred recently between some of the phages in Nature. We also show that intron homing can be highly efficient between related phages if two phages infect the same bacterium but that there also exists counteracting mechanisms that can restrict the spread of introns between phages. </p><p>In the second paper we have looked at how the presence of introns can affect gene expression in the phage. We find that the efficiency of splicing can be affected by variation of translation of the upstream exon for all three introns in T4. Furthermore, we find that splicing is also compromised upon infection of stationary-phase bacteria. This is the first time that the efficiency of self-splicing of group I introns has been coupled to environmental conditions and the potential effect of this on phage viability is discussed.</p><p>In the third paper we have characterised two novel freestanding homing endonucleases that in some T-even-like phages replace two of the putative HEGs in T4. We also present a new theory on why it is a selective advantage for freestanding, phage homing endonucleases to cleave both HEG-containing and HEG-less genomes.</p>

Visually guided navigation represents a long standing goal in robotics. Insights may be drawn from various insect species for which visual information has been shown sufficient for navigation in complex environments, however the generality of visual homing abilities across insect species remains unclear. Furthermore variousmodels have been proposed as strategies employed by navigating insects yet comparative studies across models and species are lacking. This work addresses these questions in two insect species not previously studied: the field cricket Gryllus bimaculatus for which almost no navigational data is available; and the European desert ant Cataglyphis velox, a relation of the African desert ant Cataglyphis bicolor which has become a model species for insect navigation studies. The ability of crickets to return to a hidden target using surrounding visual cues was tested using an analogue of the Morris water-maze, a standard paradigm for spatial memory testing in rodents. Crickets learned to re-locate the hidden target using the provided visual cues, with the best performance recorded when a natural image was provided as stimulus rather than clearly identifiable landmarks. The role of vision in navigation was also observed for desert ants within their natural habitat. Foraging ants formed individual, idiosyncratic, visually guided routes through their cluttered surroundings as has been reported in other ant species inhabiting similar environments. In the absence of other cues ants recalled their route even when displaced along their path indicating that ants recall previously visited places rather than a sequence of manoeuvres. Image databases were collected within the environments experienced by the insects using custompanoramic cameras that approximated the insect eye viewof the world. Six biologically plausible visual homing models were implemented and their performance assessed across experimental conditions. The models were first assessed on their ability to replicate the relative performance across the various visual surrounds in which crickets were tested. That is, best performance was sought with the natural scene, followed by blank walls and then the distinct landmarks. Only two models were able to reproduce the pattern of results observed in crickets: pixel-wise image difference with RunDown and the centre of mass average landmark vector. The efficacy of models was then assessed across locations in the ant habitat. A 3D world was generated from the captured images providing noise free and high spatial resolution images asmodel input. Best performancewas found for optic flow and image difference based models. However in many locations the centre of mass average landmark vector failed to provide reliable guidance. This work shows that two previously unstudied insect species can navigate using surrounding visual cues alone. Moreover six biologically plausible models of visual navigation were assessed in the same environments as the insects and only an image difference based model succeeded in all experimental conditions.

Homing endonucleases are rare-cutting enzymes that cleave DNA at a site near their own location, preferentially in alleles lacking the homing endonuclease gene (HEG). By cleaving HEG-less alleles the homing endonuclease can mediate the transfer of its own gene to the cleaved site via a process called homing, involving double strand break repair. Via homing, HEGs are efficiently transferred into new genomes when horizontal exchange of DNA occurs between organisms. Group I introns are intervening sequences that can catalyse their own excision from the unprocessed transcript without the need of any proteins. They are widespread, occurring both in eukaryotes and prokaryotes and in their viruses. Many group I introns encode a HEG within them that confers mobility also to the intron and mediates the combined transfer of the intron/HEG to intronless alleles via homing. Bacteriophage T4 contains three such group I introns and at least 12 freestanding HEGs in its genome. The majority of phages besides T4 do not contain any introns, and freestanding HEGs are also scarcely represented among other phages. In the first paper we looked into why group I introns are so rare in phages related to T4 in spite of the fact that they can spread between phages via homing. We have identified the first phage besides T4 that contains all three T-even introns and also shown that homing of at least one of the introns has occurred recently between some of the phages in Nature. We also show that intron homing can be highly efficient between related phages if two phages infect the same bacterium but that there also exists counteracting mechanisms that can restrict the spread of introns between phages. In the second paper we have looked at how the presence of introns can affect gene expression in the phage. We find that the efficiency of splicing can be affected by variation of translation of the upstream exon for all three introns in T4. Furthermore, we find that splicing is also compromised upon infection of stationary-phase bacteria. This is the first time that the efficiency of self-splicing of group I introns has been coupled to environmental conditions and the potential effect of this on phage viability is discussed. In the third paper we have characterised two novel freestanding homing endonucleases that in some T-even-like phages replace two of the putative HEGs in T4. We also present a new theory on why it is a selective advantage for freestanding, phage homing endonucleases to cleave both HEG-containing and HEG-less genomes.