Bradley N. White

Realized Reproductive Success of Polygynous Red-Winged Blackbirds Revealed by DNA Markers

Hypervariable genetic markers, including a novel locus-specific marker detected by a mouse major histocompatibility complex probe, reveal that multiple paternity is common in families of polygynous red-winged blackbirds (Agelaius phoeniceus). Almost half of all nests contained at least one chick resulting from an extra-pair fertilization, usually by a neighboring male. Genetically based measures of reproductive success show that individual males realize more than 20% of their overall success from extra-pair fertilizations, on average, and that this form of mating behavior confounds traditional measures of male success. The importance of alternative reproductive tactics in a polygynous bird is quantified, and the results challenge previous explanations for the evolution of avian polygny.

A RELIABLE MOLECULAR METHOD OF GENDER DETERMINATION FOR MAMMALS

Abstract A polymerase chain reaction–based method was developed for gender determination in a wide variety of marine and terrestrial mammals, including cetaceans, pinnipeds, ungulates, canids, and ursids. An intron within the zinc-finger x (Zfx) and zinc-finger y (Zfy) genes was amplified. Size variation between the Zfx and Zfy introns results in a Y-specific band in males. Size of the Zfy intron varies more across species than that of the Zfx intron. This technique is simple and rapid, requires small amounts of DNA, and can be applied to a variety of mammalian species. In contrast to previous methods, only 1 primer set is needed for both gender determination and internal verification of amplification.

Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15

We have isolated a repetitive 1.8 kb Kpnl DNA sequence which is amplified in the homogeneously staining regions of a human melanoma cell line. Under low stringency conditions this sequence (D15Z1) hybridized in situ to the centromeric heterochromatin of chromosomes 1, 9, 15p, 16, and distal Yq as well as to the the short arms of the other acrocentric chromosomes. Under conditions of high stringency, labelling was predominantly on the short arm of chromosome 15. D15Z1 was shown to be present at approximately 3,000 copies per haploid genome and organized in long tandem arrays showing restriction site heterogeneity. Sequences homologous to D15Z1 were highly enriched in the less dense shoulder region of a Ag+—Cs2SO4 gradient. Analysis of D15Z1 indicated that this sequence is composed of tandemly arranged imperfect repeats of the consensus 5′ AATGG 3′ similar to previously identified satellite III sequences. Digestion of D15Z1 with HinfI resulted in a series of restriction fragments making up a subset of the HinfI ladder components of satellites III and IV. These data suggest that D15Z1 represents a chromosome 15 specific domain of human satellites III or IV and that it makes up the major fraction of the heterochromatin of this chromosome. Possible relationships between this sequence and the cytochemical staining properties of human chromosomes with distamycin A/DAPI, D280/170, and antiserum to 5-methylcytosine are discussed.

Differential permeability of rivers to raccoon gene flow corresponds to rabies incidence in Ontario, Canada

The correlation of landscape features with genetic discontinuities reveals barriers to dispersal that can contribute to understanding present and future spread of wildlife diseases. This knowledge can then be used for targeting control efforts. The impact of natural barriers on raccoon dispersal was assessed through genetic analysis of samples from two regions, Niagara (N = 666) and St. Lawrence (N = 802). These areas are transected by major rivers and are at the northern front of a raccoon rabies epizootic. Genetic clusters were identified in each region using Bayesian clustering algorithms. In the Niagara region, two clusters were identified corresponding to either side of the Niagara River. For the St. Lawrence region, spatially congruent clusters were not identified, despite the presence of the intervening St. Lawrence River. These genetic data are consistent with raccoon rabies incidence data where rabies has been detected across the St. Lawrence River in Ontario while no cases have been detected in Ontario across the Niagara River. This is despite expectations of rabies incidence in Niagara before the St. Lawrence based on the progression of rabies from New York. The results from the two regions suggest different permeabilities to raccoons between New York and Ontario that may be attributed to the rivers. However, other factors have also been explored that could contribute to this difference between these study sites including the shape of the landscape and resource distribution.

Double in situ hybridization in combination with digital image analysis: a new approach to study interphase chromosome topography

Double in situ hybridization with mercurated and biotinylated chromosome specific DNA probes in combination with digital image analysis provides a new approach to compare the distribution of homologous and nonhomologous chromosome targets within individual interphase nuclei. Here we have used two DNA probes representing tandemly repeated sequences specific for the constitutive heterochromatin of the human chromosomes 1 and 15, respectively, and studied the relative arrangements of these chromosome targets in interphase nuclei of human lymphocytes, amniotic fluid cells, and fibroblasts, cultivated in vitro. We have developed a 2D-image analysis approach which allows the rapid evaluation of large numbers of interphase nuclei. Models to test for a random versus nonrandom distribution of chromosome segments are discussed taking into account the three-dimensional origin of the evaluated 2D-distribution. In all three human diploid cell types the measurements of target-target and target-center distances in the 2D-nuclear image revealed that the labeled segments of the two chromosomes 15 were distributed both significantly closer to each other and closer to the center of the nuclear image than the labeled chromosome 1 segments. This result can be explained by the association of nucleolus organizer regions on the short arm of chromosome 15 with nucleoli located more centrally in these nuclei and does not provide evidence for a homologous association per se. In contrast, evaluation of the interphase positioning of the two chromosome 1 segments fits the random expectation in amniotic fluid and fibroblast cells, while in experiments using lymphocytes a slight excess of larger distances between these homologous targets was occasionally observed. 2D-distances between the labeled chromosome 1 and 15 segments showed a large variability in their relative positioning. In conclusion our data do not support the idea of a strict and permanent association of these homologous and nonhomologous targets in the cell types studied so far.

Detecting recent speciation events: the case of the finless porpoise (genus Neophocaena )

Recent speciation events provide important insights into the understanding and conservation of Earths biodiversity, representing recent adaptations to a changing environment and an important source of future evolutionary potential. However, the most frequently applied criterion for molecular-based speciation investigations, that of reciprocal monophyly of mitochondrial sequences, overlooks recent speciation events where insufficient time has passed for fixed molecular differences to develop between putative species. Two morphologically distinguishable forms of finless porpoise (genus Neophocaena) exist in sympatry in the strait of Taiwan, however the taxonomic relationship of these different forms is controversial. To test the hypothesis that the two forms represent different species, a study was conducted based on morphological characters and microsatellite and mitochondrial markers. The data suggest that the two forms are highly differentiated in terms of both morphology and genetic characteristics, despite being sympatric, and therefore represent different species as defined by the biological species concept. Moreover, the two forms appear to have been reproductively isolated since sharing a common ancestor prior to the last major glaciation event ∼18 000 years ago. However, this represents an insufficient amount of time for reciprocal monophyly to have developed, and thus previous studies based on this criterion have overlooked this speciation event and resulted in incorrect taxonomic classification of these forms.

Isolation and characterization of an α-satellite repeated sequence from human chromosome 22

We constructed a library in λIL47.1 with DNA isolated from flow-sorted human chromosome 22. Over 50% of the recombinants contained the same highly repetitive sequence. When this sequence was used to probe Southern blots of EcoRI-digested genomic DNA, a ladder of bands with increments of about 170 bp was observed. This sequence comigrates with satellite III in Ag+/Cs2SO4 gradients and may account for at least part of the 170 bp Hae III ladder seen in isolated satellite III DNA. Partial sequence analysis revealed homology to the 171 bp monomeric repeat unit of α-R1-DNA and the X specific α-satellite consensus sequence. After low stringency in situ hybridization, silver grains were found over the centromeres of a number of chromosomes. Under high stringency conditions, however, the labeling was concentrated over the centromeric region of chromosome 22. This localization was confirmed using DNA from a panel of human/hamster cell lines which showed that the homologous 2.1 and 2.8 kb EcoR1 restriction fragments were chromosome 22 specific. These clones therefore contain chromosome 22 derived α-satellite sequences analogous to other chromosome-specific satellite sequences described previously.

Genetic analysis of historic western Great Lakes region wolf samples reveals early Canis lupus/lycaon hybridization

The genetic status of wolves in the western Great Lakes region has received increased attention following the decision to remove them from protection under the US Endangered Species Act. A recent study of mitochondrial DNA has suggested that the recovered wolf population is not genetically representative of the historic population. We present microsatellite genotype data on three historic samples and compare them with extant populations, and interpret published genetic data to show that the pre-recovery population was admixed over a century ago by eastern wolf (Canis lycaon) and grey wolf (Canis lupus) hybridization. The DNA profiles of the historic samples are similar to those of extant animals in the region, suggesting that the current Great Lakes wolves are representative of the historic population.

An Evolutionary Model for the Insect Vitellins

Insects can be divided into three groups based on the sizes of the polypeptide constituents of their vitellogenins and vitellins. In order to determine the relationships between these groups, antisera to the vitellins of seven insects from six taxonomic orders were used to assess immunological cross-reactivity. Antigenic relatedness was observed only between vitellins from species within the same family. Amino acid compositional data for vitellins from nine species were used to assess homology by difference matrices. The SΔQ values were similar for both intra-order and inter-order comparisons and strongly suggested relatedness. The SΔn comparisons supported the immunological data that indicated that the vitellins were evolving rapidly. For most insect vitellins there are two distinct size classes of polypeptides that seem to be derived from a single asymmetric proteolytic cleavage of a precursor. We propose a model that suggests that the different size polypeptides represent distinct domains and that in the evolution of the vitellogenin genes of the Diptera and Hymenoptera there has been domain elimination.

Combining direct and indirect genetic methods to estimate dispersal for informing wildlife disease management decisions

Epidemiological models are useful tools for management to predict and control wildlife disease outbreaks. Dispersal behaviours of the vector are critical in determining patterns of disease spread, and key variables in epidemiological models, yet they are difficult to measure. Raccoon rabies is enzootic over the eastern seaboard of North America and management actions to control its spread are costly. Understanding dispersal behaviours of raccoons can contribute to refining management protocols to reduce economic impacts. Here, estimates of dispersal were obtained through parentage and spatial genetic analyses of raccoons in two areas at the front of the raccoon rabies epizootic in Ontario; Niagara (N = 296) and St Lawrence (N = 593). Parentage analysis indicated the dispersal distance distribution is highly positively skewed with 85% of raccoons, both male and female, moving < 3 km. The tail of this distribution indicated a small proportion (< 4%) moves more than 20 km. Analysis of spatial genetic structure provided a similar assessment as the spatial genetic correlation coefficient dropped sharply after 1 km. Directionality of dispersal would have important implications for control actions; however, evidence of directional bias was not found. Separating the data into age and sex classes the spatial genetic analyses detected female philopatry. Dispersal distances differed significantly between juveniles and adults, while juveniles in the Niagara region were significantly more related to each other than adults were to each other. Factors that may contribute to these differences include kin association, and spring dispersal. Changes to the timing and area covered by rabies control operations in Ontario are indicated based on these dispersal data.