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PubMed1995

The beta-tubulin gene family evolution in the Drosophila montium subgroup of the melanogaster species group.

The beta 1-, beta 2-, and beta 3-tubulin genes have been mapped by in situ hybridization on the polytene chromosomes of 11 selected species (15 strains) belonging to the Drosophila montium subgroup. Although the hybridization pattern among the strains of the same species does not differ, this pattern is significantly different among the species. The beta-tubulin genes in the montium subgroup seem to be organized in a cluster, or in a semicluster or are completely dispersed. The clustered arrangements is found in the North-Oriental sibling species D. auraria, D. triauraria, and D. quadraria. The semiclustered arrangement, wherein the beta 1 and beta 2 genes are located at the same locus while beta 3 is at a different one, appears in the South-Oriental species D. bicornuta, D. serrata, and D. birchii, as well as in the Afrotropical species D. diplacantha and D. seguyi. The complete separation of the genes is observed in the Indian species D. kikkawai and D. jambulina and in the Afrotropical species D. vulcana. Based on the above results, a possible mode of evolution of the beta-tubulin genes in the montium subgroup is attempted. In addition, phylogenetic relationships among the montium species are discussed.

Animals↗
PubMed1993

The ecology and behaviour of the forest form of Anopheles gambiae s.s.

Studies on the ecology of Anopheles gambiae s.s. and the transmission of malaria were undertaken in a high rainfall forested area in southern Sierra Leone. Anopheles gambiae s.s., identified by chromosomal techniques as the Forest form, was the only malaria vector in the study village. Surprisingly, rice fields or swamps were not favoured breeding places for this species; breeding mainly occurred in temporary pools. The mean annual sporozoite rate of An. gambiae s.s. determined by ELISA was 3.9%. Pyrethrum spray, human bait, and exit trap collections, as well as identification of mosquito blood-meals using the ELISA method, showed that the forest chromosomal form of An. gambiae s.s. was highly anthropophagic and exophilic.

Animals↗
PubMed1996

Segregation distortion in Drosophila melanogaster: genomic organization of Responder sequences.

The heterochromatic Responder (Rsp) locus of Drosophila melanogaster is the target of the two distorter loci Sd and E(SD). Rsp is located in a specific heterochromatic region of the second chromosome and is made up of AT-rich satellite sequences whose abundance is related to its sensitivity to the distorter chromosomes. Here we report that a cluster of Rsp sequences is also located in the third chromosome. The third-chromosome cluster has the same flanking sequences as the clone originally used to identify the Rsp elements, and one of the flanking sequences is a rearranged 412 retrotransposon. The presence of a second, unlinked Rsp-sequence cluster makes re-interpretation necessary for some earlier experiments in which segregation of the third chromosome had not been followed and raises interesting possibilities for the origin of the Rsp locus.

Animals↗
PubMedDate not supplied

[Cytogenetic study of the mutagenic properties of the repellents dimethyl phthalate and phenoxyacetic acid N,N-diethylamide].

Animal experiments ascertained that the repellent N,N-diethylamide of phenoxyacetic acid (P-203) increases the frequency of chromosomal aberration in the bone marrow cells of mice following its intraperitoneal introduction and also in the cells of a regenerating liver of rats after its repeated skin application. Dimethylphthalate displayed a mutagenic action only with respect to the rats' hepatocytes when applied repeatedly to the skin.

Administration, Topical↗
PubMed1998

The mutation rate and the distribution of mutational effects of viability and fitness in Drosophila melanogaster.

The empirical distributions of the average viability and fitness of mutation accumulation lines of Drosophila melanogaster were analyzed using minimum distance estimation. Data come from two different experimental designs where mutations were allowed to accumulate: 1) in copies of chromosome II protected from natural selection and recombination (viability: Mukai et al., 1972; Ohnishi, 1977; fitness: Houle et al., 1992), 2) in inbred lines derived from the same isogenic stock (viability: Fernández & López-Fanjul, 1996; fitness: this paper). Information from all data sets converged, indicating that the mutational rates were small, about 1% for viability and 3% for fitness. For both traits, the rate of mutational decline appears to be smaller than suggested by previous studies (about one-fifth of the latter), the average mutational effect was neither severe nor very slight, ranging from -0.1 to -0.3, and the distribution of mutant effects was, at most, slightly leptokurtic. Therefore, the mutational load in natural populations is one to two orders of magnitude smaller than previously thought (as based upon analyses conditional to estimates of the mutational decline of viability or fitness that appear to be biased upward). Over 95% of the mutational variance of each trait was contributed by non-slightly deleterious mutations (absolute homozygous effect larger than 0.03 or 0.1, depending on the data set considered) occurring at a rate not higher than 0.025 per haploid genome and generation. Our data suggest that most deleterious mutations affecting fitness act mainly through a single component-trait.

Animals↗
PubMed1997

Partial sequence of the mitochondrial genome of the crustacean Daphnia pulex.

A 4062-nucleotide (nt) fragment of the mitochondrial genome of the crustacean Daphnia pulex was sequenced and found to contain the complete genes for eight tRNAs and five proteins (ATP6, ATP8, COII, COIII, ND3) and the partial sequence of COI. In combination with data described previously, approximately 50% of the D. pulex mitochondrial genome has been sequenced. The gene order in this half of the genome is identical to that of Drosophila yakuba which differs from that of the other completely sequenced crustacean mitochondrial genome, Artemia franciscana. Comparison of seven mitochondrial proteins among D. pulex, A. franciscana, D. yakuba, Anopheles gambiae, Locusta migratoria and Apis mellifera showed that, with one exception, the D. pulex proteins are most similar in length and sequence to the proteins of the dipteran insects. Conversely, patterns of nucleotide bias at third codon positions in fourfold degenerate codon families are very similar in the two crustaceans but differed substantially from the insects.

Amino Acid Sequence↗
PubMed1998

Courtship and visual defects of cacophony mutants reveal functional complexity of a calcium-channel alpha1 subunit in Drosophila.

We show by molecular analysis of behavioral and physiological mutants that the Drosophila Dmca1A calcium-channel alpha1 subunit is encoded by the cacophony (cac) gene and that nightblind-A and lethal(1)L13 mutations are allelic to cac with respect to an expanded array of behavioral and physiological phenotypes associated with this gene. The cacS mutant, which exhibits defects in the patterning of courtship lovesong and a newly revealed but subtle abnormality in visual physiology, is mutated such that a highly conserved phenylalanine (in one of the quasi-homologous intrapolypeptide regions called IIIS6) is replaced by isoleucine. The cacH18 mutant exhibits defects in visual physiology (including complete unresponsiveness to light in certain genetic combinations) and visually mediated behaviors; this mutant (originally nbAH18) has a stop codon in an alternative exon (within the cac ORF), which is differentially expressed in the eye. Analysis of the various courtship and visual phenotypes associated with this array of cac mutants demonstrates that Dmca1A calcium channels mediate multiple, separable biological functions; these correlate in part with transcript diversity generated via alternative splicing.

Alleles↗
PubMed1994

Linkage analysis of sex determination in the honey bee (Apis mellifera).

A colony-level phenotype was used to map the major sex determination locus (designated X) in the honey bee (Apis mellifera). Individual queen bees (reproductive females) were mated to single drones (fertile males) by instrumental insemination. Haploid drone progeny of an F1 queen were each backcrossed to daughter queens from one of the parental lines. Ninety-eight of the resulting colonies containing back-cross progeny were evaluated for the trait 'low brood-viability' resulting from the production of diploid drones that were homozygous at X. DNA samples from the haploid drone fathers of these colonies were used individually in polymerase chain reactions (PCR) with 10-base primers. These reactions generated random amplified polymorphic DNA (RAPD) markers that were analyzed for cosegregation with the colony-level phenotype. One RAPD marker allele was shared by 22 of 25 drones that fathered low brood-viability colonies. The RAPD marker fragment was cloned and partially sequenced. Two primers were designed that define a sequence-tagged site (STS) for this locus. The primers amplified DNA marker fragments that cosegregated with the original RAPD marker. In order to more precisely estimate the linkage between X and the STS locus, another group of bees consisting of progeny from one of the low-brood viability colonies was used in segregation analysis. Four diploid drones and 181 of their diploid sisters (workers, nonfertile females) were tested for segregation of the RAPD and STS markers. The cosegregating RAPD and STS markers were codominant due to the occurrence of fragment-length alleles. The four diploid drones were homozygous for these markers but only three of the 181 workers were homozygotes (recombinants). Therefore the distance between X and the STS locus was estimated at 1.6 cM. An additional linked marker was found that was 6.6 cM from the STS locus.

Animals↗
PubMed2005

Karyotype of Brazilian Anopheles albitarsis sensu lato (Diptera:Culicidae).

Anopheles (Nyssorhynchus) albitarsis sensu lato is an important malaria vector in Brazil, especially in the Brazilian Amazon region. Chromosome preparations of fourth-instar larvae of A. albitarsis from Iranduba and Coari (AM) and Ilha Comprida (SP) were analyzed for karyotype determination and to improve cytogenetic identification of this species. Anopheles albitarsis possesses 2n = 6 chromosomes, with two pairs (submetacentric and metacentric) of autosomes and one pair of sex chromosomes, with X-Y dimorphism. The sex pair is homomorphic and acrocentric in females and heteromorphic in males, with a punctiform Y chromosome. Somatic pairing was detected in the prometaphase and metaphase chromosomes of the three A. albitarsis populations. Apparently, sex chromosome evolution in the Culicidae does not function as does evolution in the Culicidae, since it occurs in the subfamily Anophelinae, which possesses heteromorphic sex chromosomes and is regarded as primitive, based on several criteria. These karyotype data on the albitarsis complex reinforce the hypothesis that sex chromosome evolution in the subfamily Anophelinae is conserved, and the variation revealed in the mean size of chromosomes in three populations indicates that selective pressure in these populations is occurring only at a genetic level.

Animals↗
PubMed1996

Identification of the sex-determining region of the Ceratitis capitata Y chromosome by deletion mapping.

In the medfly Ceratitis capitata, the Y chromosome is responsible for determining the male sex. We have mapped the region containing the relevant factor through the analysis of Y-autosome translocations using fluorescence in situ hybridization with two different probes. One probe, the clone pY114, contains repetitive, Y-specific DNA sequences from C. capitata, while the second clone, pDh2-H8, consists of ribosomal DNA sequences from Drosophila hydei. Clone pY114 labeled most of the long arm and pDh2-H8 hybridizes to the short arm and the centromeric region of the long arm. In 12 of the analyzed 19 Y-autosome translocation strains, adjacent-1 segregation products survive to the late pupal or even adult stage and can, therefore, be sexed. This was correlated with the length of the Y fragment still present in these aberrant individuals and allowed us to map the male-determining factor to a region of the long arm representing approximately 15% of the entire Y chromosome. No additional factors, affecting for example fertility, were detected outside the male-determining region.

Animals↗
PubMed2003

Inferences on the evolutionary history of the Drosophila americana polymorphic X/4 fusion from patterns of polymorphism at the X-linked paralytic and elav genes.

In Drosophila there is limited evidence on the nature of evolutionary forces affecting chromosomal arrangements other than inversions. The study of the X/4 fusion polymorphism of Drosophila americana is thus of interest. Polymorphism patterns at the paralytic (para) gene, located at the base of the X chromosome, suggest that there is suppressed crossing over in this region between fusion and nonfusion chromosomes but not within fusion and nonfusion chromosomes. These data are thus compatible with previous claims that within fusion chromosomes the amino acid clines found at fused1 (also located at the base of the X chromosome) are likely maintained by local selection. The para data set also suggests a young age of the X/4 fusion. Polymorphism data on para and elav (located at the middle region of the X chromosome) suggest that there is no population structure other than that caused by the X/4 fusion itself. These findings are therefore compatible with previous claims that selection maintains the strong association observed between the methionine/threonine variants at fused1 and the status of the X chromosome as fused or unfused to the fourth chromosome.

Aldehyde Dehydrogenase↗
PubMed2006

Repetitive DNA in tropical tasar silkworm Antheraea mylitta.

Antheraea mylitta is an endemic insect species producing the world famous tasar silk. Its populations occupying different ecological and geographical regions show certain degree of phenotypic variability for which they are known as 'eco-races'. In order to understand the genetic variability and phylogenetic relationship among the different eco-races we characterized a repetitive TaqI genomic DNA fragment as a genetic marker. The sequence analysis and Southern hybridization show the repetitive nature of TaqI DNA fragment, designated as A. mylitta TaqI family repeat, AmTFR. The PCR amplification of AmTFR reveals its presence in all the tested eco-races of tasar silkworm and some other silk producing insects. The AmTFR is evenly distributed in all 31 meiotic metaphase I chromosomes as observed by fluorescent in situ hybridization. The AmTFR based phylogenetic analysis of the eco-races is not congruent with the morphological variations and their geographical distribution.

Animals↗
PubMed2004

[Expression of human follicle stimulating hormone in insect cells].

To study the expression of hFSH in insect cells, the cDNA encoding the hFSH beta chain was cloned by overlapping-PCR using human chromosome DNA extracted from placental tissue as template. Then we constructed expression vector pVL1393/hFSH beta using an unfused protein nuclear polyhedrosis virus (AcNPV) expression vector. The insect cells (SF9) were cotransfected with the expression vector and nuclear polyhedrosis linearized virus DNA, and recombinant viruses AcNPV-hFSH beta were collected. The beta subunit of hFSH expressed in plasma of the SF9 cells was detected by Western blot analysis, and showed apparent molecular masses of 21 kDa. After coinfecting SF9 cells with recombinant viruses AcNPV-hFSH beta and AcNPV-hCG alpha, secreted heterodimer of hFSH was detected by Western blot under non-reducing conditions. The apparent molecular weight of heterodimer was about 33 kDa.

Animals↗
PubMed1997

Telomere maintenance without telomerase.

Telomeres are nucleoprotein structures at the ends of eukaryotic chromosomes that perform a number of vital functions. They allow a cell to distinguish between natural chromosome ends and chromosome breaks in order to delay the cell cycle and repair the broken end. Telomeres also compensate for the inability of DNA polymerase to replicate the chromosome completely. In most eukaryotes a special reverse transcriptase, telomerase, adds telomeric DNA repeats to the chromosome ends using an internal RNA template. However, evidence is accumulating for alternative elongation mechanisms in a variety of eukaryotes. In the yeast Saccharomyces cerevisiae, and possibly in humans, both of which normally use telomerase, a different mechanism can be used for chromosome length maintenance when telomerase is inactive or inactivated. Yeast apparently uses recombination for this purpose; the mechanism in humans is not known. Some insect and plant species, on the other hand, do not use telomerase as their primary mechanism for maintaining chromosome length. Drosophila makes use of specific retrotransposons for this purpose, while other dipterans use recombination. We summarize here the current knowledge of these alternative telomere elongation mechanisms.

Animals↗
PubMed1998

Analysis of a mosquito acetylcholinesterase gene promoter.

Insect acetylcholinesterase is the target site for organophosphorus and carbamate insecticides and point mutations in the Ace gene are associated with resistance in Drosophila melanogaster and Musca domestica. However, little is known of the genetic regulation of insect Ace genes. Here we report the isolation of four different cDNAs from an Aedes Ace locus and identification of the gene promoter. Northern analysis reveals two large (>10 kb) transcripts and one smaller transcript of 4 kb. The region containing the initiation of transcription was localized by sequencing the two 5' most cDNAs and by 5' RACE. The transcription start point was subsequently identified by primer extension and is flanked by a perfect arthropod initiator consensus sequence. The promoter lacks a TATA box but contains several matches to other consensus sequences for eukaryotic transcription factors. In common with the Drosophila Ace gene, there are also multiple potential initiators of translation (ATGs) upstream of the main open reading frame. The structure of the 5' leader and promoter is compared to that found in other insect and vertebrate Ace genes and the possibility that this locus is homologous to one of two Ace loci described in another mosquito, Culex pipiens, is discussed.

Acetylcholinesterase↗
PubMed2000

DNA structures common for chironomid telomeres terminating with complex repeats.

Tandem repeats, 340 bp long, have been shown to terminate the chromosomes in Chironomus pallidivittatus and similar DNA may be used for this purpose by related insects. In view of the importance of Chironomus in telomere studies, representing in principle a third system after short repeats and Drosophila telomeric retrotransposons, we have investigated the related Chironomus dilutus, to learn what DNA structures are conserved at the chromosome ends. Interspersed subrepeats in the telomeric repeats, which contain a long palindrome, and a zone of about 100 bp of relatively constant subtelomeric DNA towards the junction to the telomeric DNA, are characteristic for C. dilutus as for previously investigated species. C. dilutus has similar subtelomeric DNA at all chromosome ends, but typical telomeric repeats in only seven of the pairs since the eighth telocentric pair contains centromere-specific repeats.

Animals↗
PubMed2004

Cytological evidence for triploid males and females in the bumblebee, Bombus terrestris.

The presence of both triploid males and females in Bombus terrestris was detected by distinct chromosome observation. These cytological features are novel among the higher Hymenopteran insects. We thus strictly applied the complementary sex determination (CSD) model previously proposed for Hymenopteran insects. Three out of 60 sibling queens that were mated with diploid males produced both triploid males and females, and founded colonies. The male to female ratio of the bees which emerged from the fertilized eggs of the queens was approximately 1 to 1. Thus we reconfirm that the sex in B. terrestris is determined by a single multi-allelic locus. The body size of the triploid males was smaller than that of the diploid and haploid males. We found hatched eggs laid by one triploid female (worker). One of these developed into a 3rd instar larva, however most of the triploid individuals were sterile.

Animals↗