Estimation of the Mutagenic Potential of 8-Oxog in Nuclear Extracts of Mouse Cells Using the “Framed Mirror” Method

We propose an improved earlier described “mirror” method for detecting in cell nuclear extracts mutations that arise in DNA during its replication due to the misincorporation of deoxyadenosine-5′-monophosphate (dAMP) opposite 7,8-dihydro-8-oxoguanine (8-oxoG). This method is based on the synthesis of a complementary chain (“mirror”) by nuclear extracts of different mice organs on a template containing 8-oxoG and dideoxycytidine residue (ddC) at the 3′‑end. The “mirror” was amplified by PCR using primers part of which was non-complementary to the template. It allowed obtaining the “framed mirror” products. The misincorporation of dAMP in “framed mirror” products forms an EcoRI restriction site. The restriction analysis of double-stranded “framed mirror” products allows a quantification of the mutation frequency in nuclear extracts. The data obtained show that the mutagenic potential of 8-oxoG markedly varied in different organs of adult mice and embryos

Call diversity in the North Pacific killer whale populations: implications for dialect evolution and population history

Although killer whale, Orcinus orca, dialects have been studied in detail in several populations, little attempt has been made to compare dialect characteristics between populations. In this study we investigated geographical variation in monophonic and biphonic calls among four resident populations from the North Pacific Ocean: Northern and Southern residents from British Columbia and Washington State, southern Alaska residents, and eastern Kamchatka residents. We tested predictions generated by the hypothesis that call variation across populations is the result of an accumulation of random errors and innovation by vertical cultural transmission. Call frequency contours were extracted and compared using a dynamic time-warping algorithm. We found that the diversity of monophonic calls was substantially higher than that of biphonic calls for all populations. Repertoire diversity appeared to be related to population size: in larger populations, monophonic calls were more diverse and biphonic calls less diverse. We suggest that the evolution of both monophonic and biphonic calls is caused by an interaction between stochastic processes and directional selection, but the relative effect of directional selection is greater for biphonic calls. Our analysis revealed no direct correlation between call repertoire similarity and geographical distance. Call diversity within predefined call categories, types and subtypes, showed a high degree of correspondence between populations. Our results indicate that dialect evolution is a complex process influenced by an interaction among directional selection, horizontal transmission and founder effects. We suggest several scenarios for how this might have arisen and the implications of these scenarios for call evolution and population history