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Since changes in chromosome morphology contribute to the knowledge of evolution as well as to chromosome dynamics, this study looks specifically at one chromosome compared in twelve different species of Perissodactyls: Equus caballus (ECA), E. przewalskii (EPR), Equus africanus somaliensis (EAF), E. asinus (EAS), E. hemionus onager (EHO), E. h. kulan (EHK), E. h. kiang (EKI), E. zebra hartmannae (EZH), E. grevyi (EGR), E. burchelli (EBU), Tapirus indicus (TIN), and Rhinoceros unicornis (RUN). While chromosome morphology studies have been done in some of the extant equids, none have followed the evolution of this chromosome, homologous to Equus caballus chromosome 20 (ECA20), which contains the major histocompatibility complex (MHC). The gene order on the chromosome arm homologous to human chromosome six in most Equidae is reversed with respect to the centromere in comparison to humans. Multicolor fluorescence in situ hybridization was used to show that four probes from ECA20 hybridized to ECA20 (control), SWA5, EAS8, EHO16, EHK14, EKI16, EZH10, EGR11, EBU13, TIN4, and one of RUN12, 14, 15, or 22. The order for the four genes in the horses, zebras, and rhinoceros were as follows: cen-EDN1-MHC-ITPR3-MUT. Hybridization to the ass and tapir chromosomes displayed a possible neocentromere formation. It is apparent the chromosome has gone through several morphological changes while undergoing speciation in the Equidae, yet the overall gene order is conserved.

2. The long range DNA organisation of the class II and class III regions in eight HLA homozygous cell lines has been analysed using PFGE. Comparison of the size of the BssHII restriction fragment observed for these cell lines and five individuals possessing one to three C4 genes, shows that the organisation of the C4 genes on each chromosome can be deduced from a single PFGE experiment. Outside of the C4 and 21-OHase loci the class III region shows a highly invariant structure, with no detectable differences in the amount of DNA present. Moreover the class III region is rich in CpG-islands, one of which has been characterised, and contains at least thirteen new genes. However, in the class II region, two differences between common haplotypes have been found. The DRw52-related haplotypes have the same DNA organisation. DR2 haplotypes possess 20-30 kb more DNA in the DRB region. DRw53 haplotypes have 100-130 kb more DNA than DRw52-related haplotypes in the region containing the DRB and DQA genes.

This thesis describes studies on the binding of peptides to the murine major histocompatibility complex (MHC) class I molecule H-2D^b (D^b). The expression of the recombinant soluble D^b molecule in Chinese hamster ovary cells and its subsequent purification by nickel affinity chromatography, gel filtration, and preparative native isoelectric focusing are reported. The product is the correct molecule, homogeneous, a dimer of dimers, and free of endogenous peptide. A novel binding assay based on the enhancement of natural tryptophan fluorescence by the binding of peptide is introduced. This assay is used to determine melting curves of the empty and peptide-loaded protein, and to measure association rate constants by stopped-flow fluorescence spectroscopy. Radioligand binding measurements of equilibrium as well as association and dissociation rate constants and their temperature dependence are reported. In agreement with earlier observations, the ratio of association and dissociation rate constants is much larger than the equilibrium association constant. Fluorescence anisotropy decay spectroscopy gives evidence for conformational alterations in the D^b molecule upon peptide binding. The data, possible errors and ways to avoid them, and mathematical models of binding are discussed to obtain an overall picture of the binding process.

Both major histocompatibility genes (MHC) and infection have been shown to influence urinary odours in mice, and mice may use MHC-associated odours to detect kin for the purpose of choosing communal nesting partners, or to avoid mating with close relatives. Additionally, mice may use infection-related odours to avoid mating with sick individuals or those genetically susceptible to infection. I tested the above theories using urine as the source of odour. In a series of odour preference experiments, employing two MHC-congenic strains of mice (B10 and B10.Q), I tested the proposition that females prefer MHC-similar female odours when choosing female relatives as nesting partners, and that females prefer MHC-disparate male odours for the purpose of disassoratative mating. I found that females did not display a preference for MHC-similar female odours or MHC-disparate male odours. However, when B10.Q male urine donors were infected with 100 L3 larvae of Heligmosomoides polygyrus (Nematoda), both strains of female displayed a significant preference for the odours of uninfected B10 males. To determine whether female odour preference for uninfected males extended to actual mate choice, I subsequently examined female mate choice in a controlled setting. Using CD-1 outbred mice, I found that females preferred to mate with uninfected males over males subclinically infected with 200 L3 of H. polygyrus , as evidenced by first ejaculation preferences. In this experiment, males were tethered, and as such, male dominance interactions were prevented. Thus, in the final experiment, to assess whether male dominance interactions influenced female mate choice, in addition to infection, I tested female mate choice in a seminatural setting, which allowed for social interactions to occur. In this study, paternity was ascertained through DNA analysis and the resulting data indicated that neither male dominance status nor infection had a substantial impact on fema / Overall, the data do not support the theory that MHC-based preferences occur through urinary odours. However, the results do substantiate previous findings regarding female preferences for the odours of uninfected males. Furthermore, data from the controlled mate choice assay indicate that female odour preference for uninfected males extends to actual mate choice. Conversely, in the seminatural setting neither dominance nor infection prevailed as factors driving female mate choice. I suggest that this may be a consequence of the structural complexity of my arenas.

Thesis (Ph.D.] -- McMaster University, 1997. / Includes bibliographical references (leaves 142-155). Also available via World Wide Web.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 3 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Thesis (Ph. D.)--Northeastern University, 2008. / Title from title page (viewed March 3, 2009). Graduate School of Arts and Sciences, Dept. of Biology. Includes bibliographical references (p. 172-190).

Thesis (Ph. D.)--University of Nevada, Reno, 2008. / "May, 2008." Includes bibliographical references (leaves 99-110). Online version available on the World Wide Web.