Onchocerca volvulus: limited heterogeneity in the nuclear and mitochondrial genomes.

West African populations of Onchocerca volvulus endemic to the rain forest and savanna bioclimes of West Africa differ in their ability to induce ocular disease in infected individuals. In recent years, both clinical- and animal-model-based studies have implicated particular parasite antigens in the development of ocular onchocerciasis. To test the hypothesis that the difference in pathogenic potential of blinding and nonblinding parasites might be reflected in qualitative differences in antigens that have been implicated in the development of ocular onchocerciasis, we compared the sequences of two parasite antigens implicated in the development of ocular disease in blinding- and nonblinding-strain parasites. The results demonstrated a high level of homogeneity between the parasite strains in these genes. The study was extended to include additional nuclear genes encoding antigens that are commonly recognized by individuals infected with O. volvulus and to the mitochondrial genome of the parasite. The results demonstrate a high degree of homogeneity in both the nuclear and the mitochondrial genomes among O. volvulus isolates collected from several different sites in Africa and in the Americas. This high degree of genetic homogeneity may reflect the passage of the parasite through a recent genetic bottleneck.

Transport of protein kinase C isoforms to the nucleus of PC12 cells by nerve growth factor: association of atypical zeta-PKC with the nuclear matrix.

In an effort to understand the role of protein kinase C (PKC) in nerve growth factor-induced differentiation, we studied the expression of PKC using isoform-specific antibodies. Western blot analysis on whole cell lysates showed that alpha,beta,gamma,epsilon,zeta, iota/lambda and mu were expressed in PC12 cells, except for theta which was absent. In nuclei obtained from control PC12 cells, small amounts of delta, epsilon, iota/lambda and zeta were detected. A computer-assisted search algorithm was used to search for the presence of bipartite nuclear targeting motifs. In classical PKC isoforms alpha,beta,gamma, two bipartite motifs were present, while atypical iota/lambda and zeta-PKC displayed one motif, whereas novel PKC isoforms did not exhibit any bipartite motif structure. Treatment of cells with differentiating doses of nerve growth factor (NGF) resulted in changes of differential magnitude for all of the nuclear PKC isoforms in response to NGF. However, little change in gamma-PKC was observed in response to NGF. This analysis indicated that other factors may contribute to transport of PKC into the nucleus, in addition to the bipartite motif itself. Atypical zeta-PKC is required for NGF-induced neurite outgrowth of PC12 cells (Coleman and Wooten: J Mol Neurosci 5:39-57, 1994). Increases in nuclear zeta-PKC were NGF dose-dependant with a concomitant decrease in cytoplasmic immunoreactivity. The localization of zeta-PKC was investigated by means of immunoelectron microscopy which revealed the localization of this isoform within the inner nuclear matrix bound to chromatin. Taken together, these findings suggest that zeta-PKC may be involved in the regulation of nuclear processes.

Recent evolutionary history of American Onchocerca volvulus, based on analysis of a tandemly repeated DNA sequence family.

Polymerase chain reaction (PCR) products were characterized for a repeated sequence family (designated "O-150") of the human filarial parasite Onchocerca volvulus. In phylogenetic inferences, the O-150 sequences clustered into closely related groups, suggesting that concerted evolution maintains sequence homology in this family. Using a novel mathematical model based on a nested application of an analysis of variance, we demonstrated that African rainforest and savannah strain parasite populations are significantly different. In contrast, parasites collected in the New World are indistinguishable from African savannah strains of O. volvulus. This finding supports the hypothesis that onchocerciasis was recently introduced into the New World, possibly as a result of the slave trade.

Spatial autocorrelation analysis of migration and selection.

We test various assumptions necessary for the interpretation of spatial autocorrelation analysis of gene frequency surfaces, using simulations of Wright's isolation-by-distance model with migration or selection superimposed. Increasing neighborhood size enhances spatial autocorrelation, which is reduced again for the largest neighborhood sizes. Spatial correlograms are independent of the mean gene frequency of the surface. Migration affects surfaces and correlograms when immigrant gene frequency differentials are substantial. Multiple directions of migration are reflected in the correlograms. Selection gradients yield clinal correlograms; other selection patterns are less clearly reflected in their correlograms. Sequential migration from different directions and at different gene frequencies can be disaggregated into component migration vectors by means of principal components analysis. This encourages analysis by such methods of gene frequency surfaces in nature. The empirical results of these findings lend support to the inference structure developed earlier for spatial autocorrelation analysis.

Reestablishment of the Perdido Key Beach Mouse (Peromyscus polionotus trissyllepsis) on Gulf Islands National Seashore.

In April 1986, the endangered Perdido Key beach mouse (Peromyscus polionotus trissyllepsis) existed only as a small population of less than 30 animals on the western end of Perdido Key at Gulf State Park, Alabama This population was vulnerable to extinction from a variety of causes. Fifteen pairs of mice from Alabama were moved approximately 20 km on the same island to Gulf Islands National Seashore Florida between November 1986 and April 1988. The Alabama population was surveyed by live-trapping before each removal and showed a large increase during this study. Eleven pairs of mice were released into enclosures to stimulate burrowing and reduce dispersal at the release site. The last four pairs w e released unrestricted into the dune habitat Trapping in July 1988 revealed that virtually all available dune habitat (11,000 linear m; approximately 160 ha) had been occupied by the mice. Fifty-five individuals were captured including four of the released mice. Exchanges between the populations are recommended to prevent loss of genetic diversity. Future research should investigate demographics, dispersal pattern, and the application of DNA fingerprinting techniques to determine rates of gene flow in the population. The Perdido Key beach mouse provides an excellent model for studying the effects of a population bottleneck on genetic diversity and testing the predictions of population viability analysis.