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A controlled and efficient coordination of gene expression is the key for normal development of an organism. In mammals, a subset of autosomal genes is expressed monoallelically depending on the sex of the transmitting parent, a phenomenon known as genomic imprinting. The imprinted state of the H19 and Igf2 genes is controlled by a short stretch of sequences upstream of H19 known as the imprinting control region (ICR). This region is differentially methylated and is responsible for the repression of the maternally inherited Igf2 allele. It harbors hypersensitive sites on the unmethylated maternal allele and functions as an insulator that binds a chromatin insulator protein CTCF. Hence the H19 ICR, which plays an important role in maintaining the imprinting status of H19 and Igf2, was shown to lose the insulator property upon CpG methylation. Another ICR in the Kcnq1 locus regulates long-range repression of p57Kip2 and Kcnq1 on the paternal allele, and is located on the neighboring subdomain of the imprinted gene cluster containing H19 and Igf2, on the distal end of mouse chromosome 7. Similarly to the H19 ICR, the Kcnq1 ICR appears to possess a unidirectional and methylation-sensitive chromatin insulator property in two different somatic cell types. Hence, methylation dependent insulator activity emerges as a common feature of imprinting control regions. The protein CTCF is required for the interpretation and propagation of the differentially methylated status of the H19 ICR. Work in this thesis shows that this feature applies genomewide. The mapping of CTCF target sites demonstrated not only a strong link between CTCF, formation of insulator complexes and maintaining methylation-free domains, but also a network of target sites that are involved in pivotal functions. The pattern of CTCF in vivo occupancy varies in a lineage-specific manner, although a small group of target sites show constitutive binding. In conclusion, the work of this thesis shows that epigenetic marks play an important role in regulating the insulator property. The studies also confirm the importance of CTCF in maintaining methylation-free domains and its role in insulator function. Our study unravels a new range of target sites for CTCF involved in divergent functions and their developmental control.

Across species, the face and more specifically the mouth, serves as an essential facet of everyday life. Amongst humans the mouth serves as a tool for the ingestion of food, a marker for facial recognition and a medium for communication. In order for the mouth to properly form, a series of precise growth and fusion events are needed. In order to insure that these events are orchestrated properly is a wide array of signals, transcription factors and epigenetic regulators. Due to the needed precision of these events, congenital birth defects of the face such as cleft lip and cleft palate are some of the most common worldwide. In order to support existing and identify new developmental processes involved in mouth formation, we have utilized the effective model, Danio to study the molecules and events implicated in orofacial development. This was accomplished by developing a novel confocal imaging technique that allows for visualization of the forward facing zebrafish. Using this imaging technique we were able to establish when the embryonic mouth first forms in zebrafish. Additionally, we recapitulated cleft-palate phenotypes shown in previous literature with the imaging method. Utilizing this technique, we then sought to further establish the role of Ca2+ signaling in proper orofacial morphogenesis and determine if the serine/threonine protein kinase, Ca2+/calmodulin-dependent protein kinase type-II (CaMK-II), has a role in proper orofacial developmental.

Among vertebrates, oviparity, a condition in which young at least partially develop outside the mother's body and are supported by yolk, is the ancestral state to viviparity, the reproductive mode in which embryos are fully developed at birth. Viviparity in reptiles is found only in the squamates. Among the more than 100 origins of viviparity in squamates, many have occurred fairly recently and as a result some species contain populations of both reproductive modes, i.e., are reproductively bimodal. The evolution of viviparity is associated with many changes including an increase in oviductal egg retention times, a decrease in thickness of the eggshell and a decrease in secretion of the uterine shell glands that are responsible for eggshell secretion. The uterine morphology of the oviparous Saproscincus mustelinus and the reproductively bimodal Saiphos equalis were compared to study the uterine shell gland cycle.

<p>For a complete picture of the lake ecosystem, it is necessary to understand the mechanisms regulating biomass and nutrient status (nutrient limitation, optimal supply, or surplus) of benthic algae, which are important primary producers and a food resource for grazers. This thesis gives an overview of the natural variation of benthic algae at different scales of space and time and on different substrates, and unravels some of the underlying factors. Algal nutrient status was assessed using the C:N:P (carbon:nitrogen:phosphorus) ratio of the entire natural benthic community. A review, observations, and experiments confirmed that a C:N:P ratio of about 158:18:1 (molar basis) represented an optimal nutrient supply, and that substantially higher C:N, N:P, or C:P ratios reflected algal growth limitation caused by an N or P nutrient deficiency. </p><p>Horizontal variation of benthic algal biomass and nutrient status was patchy, of similar amount for all investigated distances, substrates, and lakes, and constituted a dominant proportion of the total variation. For example, patches of nutrient limited algae were found within only 10 m distance from patches with a nutrient surplus. Thus, horizontal variation should not be neglected when sampling benthic algae in lakes. Field observations suggested an impact of wind, nutrients, and grazers on the horizontal variation. Light and nutrients might have caused the observed vertical and temporal variation. Field experiments confirmed a simultaneous control of benthic algal biomass by nutrients and grazing, mediated by light and temperature. Grazing effects were larger than nutrient effects, but the comparison of natural communities in lakes of different trophy suggested that benthic algal biomass was controlled by nutrients in the long run. </p><p>An important nutrient supply was animal excretions, causing a low C:N:P ratio of epizoon on zebra mussels, and algal communities associated with macrograzers. A field experiment revealed that 15N circulated one week longer in epizoon associated with a sessile caddisfly than in surrounding epilithon. In conclusion, the regulation of benthic algal biomass and nutrient status in lakes is complex, and benthic animals should be looked at not only as grazers, but also as a nutrient source.</p>