Tracking changes in chromosomal arrangements and their genetic content during adaptation.

There is considerable evidence for an adaptive role of inversions, but how their genetic content evolves and affects the subsequent evolution of chromosomal polymorphism remains controversial. Here, we track how life-history traits, chromosomal arrangements and 22 microsatellites, within and outside inversions, change in three replicated populations of Drosophila subobscura for 30 generations of laboratory evolution since founding from the wild. The dynamics of fitness-related traits indicated adaptation to the new environment concomitant with directional evolution of chromosomal polymorphism. Evidence of selective changes in frequency of inversions was obtained for seven of 23 chromosomal arrangements, corroborating a role for inversions in adaptation. The evolution of linkage disequilibrium between some microsatellites and chromosomes suggested that adaptive changes in arrangements involved changes in their genetic content. Several microsatellite alleles increased in frequency more than expected by drift in targeted inversions in all replicate populations. In particular, there were signs of selection in the O3+4 arrangement favouring a combination of alleles in two loci linked to the inversion and changing along with it, although the lack of linkage disequilibrium between these loci precludes epistatic selection. Seven other alleles increased in frequency within inversions more than expected by drift, but were not in linkage disequilibrium with them. Possibly these alleles were hitchhiking along with alleles under selection that were not specific to those inversions. Overall, the selection detected on the genetic content of inversions, despite limited coverage of the genome, suggests that genetic changes within inversions play an important role in adaptation.

Hsp70 protein levels and thermotolerance in Drosophila subobscura: a reassessment of the thermal co-adaptation hypothesis.

Theory predicts that geographic variation in traits and genes associated with climatic adaptation may be initially driven by the correlated evolution of thermal preference and thermal sensitivity. This assumes that an organism's preferred body temperature corresponds with the thermal optimum in which performance is maximized; hence, shifts in thermal preferences affect the subsequent evolution of thermal-related traits. Drosophila subobscura evolved worldwide latitudinal clines in several traits including chromosome inversion frequencies, with some polymorphic inversions being apparently associated with thermal preference and thermal tolerance. Here we show that flies carrying the warm-climate chromosome arrangement O(3+4) have higher basal protein levels of Hsp70 than their cold-climate O(st) counterparts, but this difference disappears after heat hardening. O(3+4) carriers are also more heat tolerant, although it is difficult to conclude from our results that this is causally linked to their higher basal levels of Hsp70. The observed patterns are consistent with the thermal co-adaptation hypothesis and suggest that the interplay between behaviour and physiology underlies latitudinal and seasonal shifts in inversion frequencies.

Contrasting patterns of phenotypic variation linked to chromosomal inversions in native and colonizing populations of Drosophila subobscura.

In fewer than two decades after invading the Americas, the fly Drosophila subobscura evolved latitudinal clines for chromosomal inversion frequencies and wing size that are parallel to the long-standing ones in native Palearctic populations. By sharp contrast, wing shape clines also evolved in the New World, but the relationship with latitude was opposite to that in the Old World. Previous work has suggested that wing trait differences among individuals are partially due to the association between chromosomal inversions and particular alleles which influence the trait under consideration. Furthermore, it is well documented that a few number of effective individuals founded the New World populations, which might have modified the biometrical effect of inversions on quantitative traits. Here we evaluate the relative contribution of chromosomal inversion clines in shaping the parallel clines in wing size and contrasting clines in wing shape in native and colonizing populations of the species. Our results reveal that inversion-size and inversion-shape associations in native and colonizing (South America) populations are generally different, probably due to the bottleneck effect. Contingent, unpredictable evolution was suggested as an explanation for the different details involved in the otherwise parallel wing size clines between Old and New World populations of D. subobscura. We challenge this assertion and conclude that contrasting wing shape clines came out as a correlated response of inversion clines that might have been predicted considering the genetic background of colonizers.

The chromosomal polymorphism of Drosophila subobscura: a microevolutionary weapon to monitor global change.

The Palaearctic species Drosophila subobscura recently invaded the west coast of Chile and North America. This invasion helped to corroborate the adaptive value of the rich chromosomal polymorphism of the species, as the same clinal patterns than those observed in the original Palaearctic area were reproduced in the colonized areas in a relatively short period of time. The rapid response of this polymorphism to environmental conditions makes it a good candidate to measure the effect of the global rising of temperatures on the genetic composition of populations. Indeed, the long-term variation of this polymorphism shows a general increase in the frequency of those inversions typical of low latitudes, with a corresponding decrease of those typical of populations closer to the poles. Although the mechanisms underlying these changes are not well understood, the system remains a valid tool to monitor the genetic impact of global warming on natural populations.

Tracking the origin of an invasive species: Drosophila subobscura in Argentina.

Biological invasions are excellent opportunities to study the evolutionary forces leading to the adaptation of a species to a new habitat. Knowledge of the introduction history of colonizing species helps tracking colonizing routes and assists in defining management strategies for invasive species. The Palearctic species Drosophila subobscura is a good model organism for tracking colonizations since it was detected in Chile and western North America three decades ago and later on in the Atlantic coast of Argentina. To unravel the origin of the Argentinean colonizers two populations have been analysed with several genetic markers. Chromosomal arrangements and microsatellite alleles found in Argentina are almost similar to those observed in Chile and USA. The lethal allelism test demonstrates that the lethal gene associated with the O(5) inversions in Argentina is identical to that found in Chile and USA, strongly supporting the hypothesis that all the American colonizing populations originated from the same colonization event. A secondary bottleneck is detected in the Argentinean populations and the genetic markers suggest that these populations originated from the invasion of 80-150 founding individuals from Chile.

Malrotación intestinal como causa de dolor abdominal crónico recurrente / Malrotación intestinal como causa de dolor abdominal

La malrotación intestinal como causa de dolor abdominal en adultos es una entidad poco frecuente. Presentamos el caso de un varón de 17 años con antecedentes de dolor abdominal crónico de 10 años de evolución, que ingresó por urgencias como consecuencia de la agudización de su dolor y al que se le practicó una laparotomía exploradora de forma urgente, encontrando una malrotación intestinal con enroscamiento en espiral del meso del intestino delgado y presencia de bandas de Ladd (AU)

The intestinal malrotation as cause of abdominal pain in adults is a not very frequent entity. We present the case of a 17 year-old male with antecedents of chronic abdominal pain of 10 years of evolution who went to the emergency room as a consequence of an increasing pain and who underwent an emergency exploratory laparotomy, thus finding an intestinal malrotation with spiral swinging of the meso of the midgut and presence of Ladd's bands (AU)

Introduction history of Drosophila subobscura in the New World: a microsatellite-based survey using ABC methods.

Drosophila subobscura is a Palearctic species that was first observed in South and North America in the early 1980s, and that rapidly invaded broad latitudinal ranges on both continents. To trace the source and history of this invasion, we obtained genotypic data on nine microsatellite loci from two South American, two North American and five European populations of D. subobscura. We analysed these data with traditional statistics as well as with an approximate Bayesian computation (ABC) framework. ABC methods yielded the strongest support for the scenario involving a serial introduction with founder events from Europe into South America, and then from South America into North America. Stable effective population size of the source population was very large (around one million individuals), and the propagule size was notably smaller for the introduction into South America (i.e. high bottleneck severity index with only a few effective founders) but considerably larger for the subsequent introduction into North America (i.e. low bottleneck severity index with around 100-150 effective founders). Finally, the Mediterranean region of Europe (and most likely Barcelona from the localities so far analysed) is proposed as the source of the New World flies, based on mean individual assignment statistics.

Swift laboratory thermal evolution of wing shape (but not size) in Drosophila subobscura and its relationship with chromosomal inversion polymorphism.

Latitudinal clinal variation in wing size and shape has evolved in North American populations of Drosophila subobscura within about 20 years since colonization. While the size cline is consistent to that found in original European populations (and globally in other Drosophila species), different parts of the wing have evolved on the two continents. This clearly suggests that 'chance and necessity' are simultaneously playing their roles in the process of adaptation. We report here rapid and consistent thermal evolution of wing shape (but not size) that apparently is at odds with that suggestion. Three replicated populations of D. subobscura derived from an outbred stock at Puerto Montt (Chile) were kept at each of three temperatures (13, 18 and 22 degrees C) for 1 year and have diverged for 27 generations at most. We used the methods of geometric morphometrics to study wing shape variation in both females and males from the thermal stocks, and rates of genetic divergence for wing shape were found to be as fast or even faster than those previously estimated for wing size on a continental scale. These shape changes did not follow a neat linear trend with temperature, and are associated with localized shifts of particular landmarks with some differences between sexes. Wing shape variables were found to differ in response to male genetic constitution for polymorphic chromosomal inversions, which strongly suggests that changes in gene arrangement frequencies as a response to temperature underlie the correlated changes in wing shape because of gene-inversion linkage disequilibria. In fact, we also suggest that the shape cline in North America likely predated the size cline and is consistent with the quite different evolutionary rates between inversion and size clines. These findings cast strong doubts on the supposed 'unpredictability' of the geographical cline for wing traits in D. subobscura North American colonizing populations.

Linkage disequilibria between mtDNA haplotypes and chromosomal arrangements in a natural population of Drosophila subobscura.

The association between mtDNA haplotypes and chromosomal arrangements in a natural population of Drosophila subobscura from Calvia (Balearic Islands, Spain) was studied in order to search for linkage disequilibria, in an attempt to explain the populational dynamics of the mtDNA haplotypes of this species in nature. The presence of Wolbachia was not detected. Two main haplotypes (I and II) were found, as well as a series of less common ones. The Tajima D-test seemed to indicate some kind of seasonal population bottleneck. An analysis of linkage disequilibrium and factorial analysis of correspondences detected an association between haplotype I and the J(ST) inversion and haplotype II and the J(1) inversion.

Colonization of America by Drosophila subobscura: heterotic effect of chromosomal arrangements revealed by the persistence of lethal genes.

About 20 years ago Drosophila subobscura, a native Palearctic species, colonized both North and South America. In Palearctic populations lethal genes are not associated in general with particular chromosomal arrangements. In colonizing populations they are not randomly distributed and usually are associated to a different degree with chromosomal arrangements caused by the founder event. The persistence of two lethal genes in the colonizing populations, one completely associated with the O(5) inversion and the other partially associated with the O(3+4+7) arrangement, has been analyzed. In all populations studied (five North American and six South American) the observed frequency of the lethal gene completely associated with the O(5) inversion is higher than expected, the difference being statistically significant in all South American and one North American populations. The observed frequency of the lethal gene partially associated with the O(3+4+7) arrangement is also significantly higher than expected. Taking into account that the O(5) inversion exhibits significant latitudinal clines both in North and South America, an overdominant model favoring the heterokaryotypes seems to be in operation. From this model, a polynomial expression has been developed that allows us to estimate the relative fitness and the coefficient of selection against all karyotypes not carrying the O(5) inversion. The relative fitness of the O(5) heterokaryotypes is higher in South American than in North American populations. Furthermore, the observed frequencies of the lethal genes studied are in general very close to those of the equilibrium. This case is an outstanding demonstration in nature of an heterotic effect of chromosomal segments associated with lethal genes on a large geographic scale.

Colonization of America by Drosophila subobscura: spatial and temporal lethal-gene allelism.

About twenty years ago Drosophila subobscura, a western Palearctic species, colonized both North and South America. Lethal genes in the O chromosome has been subject to much research. Lethal gene allelisms between American populations far away have been studied. These allelisms were not negligible, but all cases were due to the lethal gene completely associated to the O5 chromosomal inversion. Here we analyze the lethal genes in a new American population of D. subobscura (Centralia, Washington), located fairly close to a previously studied population (Bellingham, Washington) and separated in space and time with other American populations (Gilroy I and II in California and Santiago de Chile). The frequencies of lethal and semilethal genes of Centralia were 16.9+/-4.6 and 6.2+/-3.0, respectively. The intrapopulational allelism of Centralia was 0.122+/-0.036. Interpopulational allelisms were studied using the lethal genes from the populations separated in space and time from Centralia. The interpopulational allelisms between Centralia and Gilroy I (California) and between Centralia and Bellingham (Washington) were higher than the intrapopulational allelism (0.155+/-0.032 and 0.153+/-0.024, respectively). In all these cases allelism was due to a complete association between a lethal gene and the O5 chromosomal inversion. Accordingly, no other lethal genes are shared in these populations.

Diagnosis of sibling species of Drosophila involved in the colonization of North America by D. subobscura.

To determine the effects of the recent colonization of the west coast of North America by the Palaearctic species Drosophila subobscura on the dynamics of the Drosophila populations, the sibling species D. athabasca and D. azteca must be classified unambiguously. We have characterized these two species using three molecular techniques: allozymes, mtDNA and RAPDs. All three techniques allow the classification of any individual as belonging to either species. The study of five localities in northern California and southern Oregon show that the area of overlap is larger than previously described.

Colonization of America by Drosophila subobscura: the founder event and a rapid expansion.

The recent colonization of the American continent by Drosophila subobscura offers a unique opportunity to analyze the response of different genomic entities to a new environment. Here, results from a study of 15 allozyme loci in six North American and seven South American populations are compared with existing data from Palearctic populations. In general, only alleles with a frequency higher than 0.1 in European populations are present in America. The observed alteration in allele frequencies can be explained by a founder event. Although some significant latitudinal clines for allozyme frequencies have been detected, the results obtained are not as clear-cut as are those for chromosomal inversions. Overall heterozygosity is similar between North and South American populations and does not differ from that of Palearctic populations. This observation experimentally corroborates the results provided by theoretical models in which the average heterozygosity per locus depends not only on the size of bottleneck but also on the rate of population growth. The resemblance between North and South American populations is clearly manifested by a geometrical representation using Bhattacharyya's distance and a multidimensional scaling technique. Furthermore, these American populations are clearly differentiated from the Palearctic populations.

In vitro oxidised HDL is recognized by the scavenger receptor of macrophages: implications for its protective role in vivo.

To assess the effects of oxidative modification, human HDL was oxidised in vitro for 12 h (Ox-HDL12) and 24 h (Ox-HDL24) under similar conditions to those commonly used for LDL. The procedure resulted in: an increase in thiobarbituric acid reactive substances but with marginal change in electronegativity; protein denaturation accounting for 16% and 45% loss of immunoreactive apoprotein A-I in the Ox-HDL12 and Ox-HDL24 respectively relative to the non-oxidised, native HDL (Nat-HDL); a decrease in the polyunsaturated fatty acids of the triglyceride, cholesterol ester and phospholipid components of the lipoprotein; an increase in the proportion of short chain saturated fatty acids while the monounsaturated fatty acids remained relatively unchanged. Studies with human macrophages demonstrated: a decrease of 16% and 30% in the capacity of the Ox-HDL12 and Ox-HDL24 respectively to efflux intracellular free cholesterol; 125I-Ox-HDL24 uptake and degradation was directly comparable with that of 125I-Ac-LDL; the addition of excess unlabelled Ox-HDL24, Ac-LDL, Ox-LDL24 and Nat-HDL resulted in 74%, 67%, 69% and 19% displacement of the 125I-Ox-HDL24 respectively; fucoidin and dextran sulphate displaced 125I-Ox-HDL by 20% and 40% respectively; intracellular free and esterified cholesterol was increased 2.5-fold and 4-fold respectively relative to Nat-HDL on incubation with Ox-HDL24. These findings suggest that HDL is susceptible to oxidative modification leading to recognition by the scavenger receptor of macrophages and subsequent intracellular cholesterol accumulation. As such, the in vivo protective role of HDL in cardiovascular disease can be reversed in those circumstances in which HDL, like LDL, undergoes oxidative modification.

[Polymorphism (RFLP-PstI) of the apoprotein A-I gene in a healthy population. Its relation to high-density lipoproteins]. / Polimorfismo (RFLP-PstI) del gen de la apoproteína A-I, en población sana. Relación con las lipoproteínas de alta densidad.

BACKGROUND: Some RFLPs for the genes encoding for apoproteins have been associated with dyslipidemia and the predisposition to atherosclerosis. It is of interest to investigate the apo A-I gene in a Mediterranean population, since it is the major protein in HDL. METHODS: We studied the A-I C-III A-IV gene cluster RFLP defined by the endonuclease Pst I in 149 healthy males randomly selected among industrial workers in Tarragona. The mean age was 40 +/- 7 years (range 20 to 62). We analysed cholesterol and triglycerides in plasma and the lipoprotein fractions (VLDL, IDL, LDL and HDL) obtained by ultracentrifugation. The RFLP was determined for the enzyme Pst I in the A-I C-III A-IV gene cluster by the Southern blotting method. RESULTS: The genotype distribution was P1P1 81.9%, P1P2 14.8% and P2P2 3.4% and the allelic frequency was P1 89.3% and P2 10.7%. The plasma cholesterol, triglycerides, apo A-I and apo B did not show significant differences between these groups. The P2P2 subjects had lower HDL-C values (P1P1 1.17 +/- 0.39 mmol/l, P1P2 1.16 +/- 0.28 mmol/l y P2P2 0.89 +/- 0.14 mmol/l; p < 0.01). CONCLUSIONS: The distribution of the genotypes in the Mediterranean population were similar to that observed in the USA and in Europe. P2P2 subjects had decreased HDL cholesterol but the low prevalence of the genotype being very low limits its value as a marker of coronary artery disease risk.

Low density lipoprotein ligand-receptor interactions in normal healthy individuals characterized by their XbaI apolipoprotein B DNA polymorphism.

A DNA restriction fragment length polymorphism (RFLP), observed with the XbaI restriction enzyme digestion of peripheral lymphocyte genomic DNA and a 3.5 kb probe 3' end of the apolipoprotein B gene, was investigated in 228 normal healthy males. Lipoprotein measurements were conducted on fasting plasma and related to the genotype; the X2X2 homozygotes (the X2 allele contains the enzyme cutting site) had significantly higher plasma cholesterol, low density (LDL) cholesterol and LDL apolipoprotein B. Thirty subjects (10 from each of the X1X1, X1X2 and X2X2 groups) were recalled and the LDL receptor activity measurements, conducted on peripheral venous blood lymphocytes, indicated no significant differences between the genotypes. However, when LDLs isolated from these individuals were assayed for ligand-receptor interaction with a human embryonic lung fibroblast cell line, significantly different maximum binding (Bmax) values in the X2 allele-bearing individuals were observed. This paradoxically elevated in vitro binding and degradation of LDL from X2X2 subjects suggests that the elevated concentrations of LDL cholesterol observed with this genotype in vivo does not result from a defective ligand-receptor interaction directly related to this polymorphism.

[Polymorphisms of the Apo B gene in a healthy population and its association with hyperlipidemia]. / Polimorfismos del gen de la Apo B en población sana y su asociación a hiperlipidemias.

The apoprotein B gene restriction fragment length polymorphism are studied using the endonuclease XbaI, and the pAB3.5C probe was studied in 128 healthy males aged 20-62 years (39.2 +/- 7.6). The genotypic prevalence was X1X1 26.6%; X1X2 47.7% and X2X2 25.7%. The allelic frequency was 50.3% X1 and 49.7 for X2. No differences in prevalence were observed related to age or body mass index. The genotype X2X2 was statistically associated with a 10% increase in total plasma cholesterol, LDL cholesterol and LDL Apo B levels (p less than 0.05). Up to 6% of the total plasma cholesterol levels were dependent on X2X2 genotype as shown by multivariate regression analysis. The X2X2 genotype may be a candidate marker in assessing increased risk for coronary heart disease.