PubMed HealthSearch

Biomedical subjects

W W Anderson

Publications and source records attributed to W W Anderson.

At least 19 recordsLinked to original sources

Evidence for gene conversion in the amylase multigene family of Drosophila pseudoobscura.

The alpha-amylase (Amy) multigene family in Drosophila pseudoobscura is located on the third chromosome, which is polymorphic for more than 40 inverted gene arrangements. The number of copies in this family ranges from one to three, depending on the arrangement in question. A previous study of the three Amy genes from the Standard (ST) arrangement suggested either that duplicated copies (Amy2 and Amy3) are functionally constrained or that they are undergoing gene conversion with Amy1. In order to elucidate further the pattern of molecular evolution in this family, we cloned and sequenced four additional Amy genes, two from the Santa Cruz (SC) and two from the Chiricahua (CH) gene arrangement. Of the two alternatives, only the hypothesis of gene conversion is supported by the sequence analysis. The homogenization effect of gene conversion has been strongest in SC, whose copies differ by only two nucleotides, less noticeable in ST, and negligible in the CH. Furthermore, the action of gene conversion is apparently localized, occurring only in the coding region. Interestingly, these results concur with the findings of other workers for the duplicated Amy genes in the Drosophila melanogaster group. Thus, the occurrence of gene conversion in the Amy multigene family seems to be a common feature in the Drosophila species studied so far.

Animals

Interchromosomal exchange of genetic information between gene arrangements on the third chromosome of Drosophila pseudoobscura.

During the last 60 years, the inversion polymorphism on the third chromosome of Drosophila pseudoobscura has become a case study of the evolution of linked blocks of genes, isolated from each other by the suppression of recombination in heterozygotes for different inversions. Due to its location within inverted regions in most gene arrangements, the amylase (Amy) gene region can be used to elucidate the molecular pattern of evolution in these inversions. We studied this region in the Tree Line phylad of gene arrangements, with regard to both restriction site polymorphisms (RSP) and nucleotide sequences. The analysis of restriction maps, encompassing 26 kb, corroborates the cytogenetic phylogeny established on the basis of inversion breakpoints. However, we found that the 2.7 kb of nucleotide sequences of the AmyI gene are identical in both Estes Park and Hidalgo arrangements, despite the fact that these inversions arose independently from Tree Line. These contrasting results suggest that a homogenizing force, most likely gene conversion, is able to bring about localized exchanges between otherwise isolated gene arrangements.

Amylases

The history of a genetic system.

Although the chromosomal polymorphism for inversions in Drosophila pseudoobscura is one of the best studied systems in population genetics, the identity of the ancestral gene arrangement has remained unresolved for more than 50 years. There are more than 40 gene arrangements, and 4 of them (Standard, Hypothetical, Santa Cruz, and Tree Line) have been considered as candidates for the ancestral type. We propose a framework of competing hypotheses to distinguish among the alternatives. Two conclusions come from contrasting each hypothesis with the results from DNA sequencing and restriction mapping. First, not only Standard but also Hypothetical can be excluded as the ancestral gene arrangement. Second, although either Tree Line or Santa Cruz could be the ancestral type, the available data provide greater support for Santa Cruz.

Amylases

Temperature related fertility selection on body size and the sex-ratio gene arrangement in Drosophila pseudoobscura.

We measured temperature-dependent fertility selection on body size in Drosophila pseudoobscura in the laboratory. One hundred single females of each of the three karyotypes involving the 'sex-ratio' (SR) and the standard (ST) gene arrangement on the sex chromosome laid eggs at either 18 or 24 degrees C. The experiment addressed the following hypotheses: (a) Fertility selection on body size is weaker at the higher temperature, explaining in part why genetically smaller flies appear to evolve in populations at warmer localities. (b) Homokaryotypic SR females are less fecund than homokaryotypic ST females, possibly mediated by the effect of body size on fertility, explaining the low frequencies of SR despite its strong advantage due to meiotic drive. The data were also expected to shed light on a mechanism for the evolution of plasticity of body size through fertility selection in environments with an unpredictable temperature regime. Hypothesis (a) was clearly refuted because phenotypically larger ST females had an even larger fertility surplus at the higher temperature and, more importantly, the genetic correlation between fertility and body size disappeared at the lower temperature. As to (b), we found that temperature affects fertility directly and indirectly through body size such that ST and SR females were about equally fecund at both temperatures, although different in size and size-adjusted fertility. We observed heterosis for both size and fertility, which might stabilize the polymorphism in nature. The reaction norms of body size to the temperature difference were steeper for ST females than for SR females, implying that fertility selection could change phenotypic plasticity of body size in a population. Selection on body size depended not only on the temperature, but also on the karyotypes, suggesting that models of phenotype evolution using purely phenotypic fitness functions may often be inadequate.

Animals

Four decades of inversion polymorphism in Drosophila pseudoobscura.

We report data that continue the studies of Dobzhansky and others on the frequencies of third-chromosome inversions in natural populations of Drosophila pseudoobscura in North America. The common gene arrangements continue to be present in frequencies similar to those described four decades ago, and the broad geographic patterns also remain unchanged. There is only one pronounced trend over time: the increase in frequency of the Tree Line inversion in Pacific coast populations.

Animals

Molecular evolution of inversions in Drosophila pseudoobscura: the amylase gene region.

The amylase region of the third chromosome of Drosophila pseudoobscura has been cloned and localized to cytological band 73A. It is contained within a series of highly polymorphic inversions and serves as a convenient tool for a molecular evolutionary analysis of the inverted gene arrangements. Amylase in D. pseudoobscura is a family of three genes, and some chromosomes have deletions for one or two of them. Two overlapping clones covering 26 kilobases were isolated and used as probes to survey DNA restriction map polymorphism among 28 lines, representing five of the major inversion types found in natural populations, as well as single chromosomes from the closely related species Drosophila persimilis and Drosophila miranda. Restriction-site differences are considerably greater among the various gene arrangements than among chromosomes with the same gene arrangement. Clustering the restriction map haplotypes yielded a dendrogram concordant with the phylogeny generated independently from cytogenetic considerations. The inversion polymorphism is estimated to be about 2 million years old.

Amylases

Exposure of hippocampal slices to magnesium-free medium produces epileptiform activity and simultaneously decreases calcium and calmodulin-dependent protein kinase II activity.

The effect of magnesium-free medium on electrical and CaM kinase II activity in the rat hippocampal slice was examined. Experimental slices were incubated in 2 mM Mg, then exposed to magnesium-free medium for 1 h. Control slices were concurrently run in 2 mM Mg. Slices were then frozen and CaM kinase II activity was measured in homogenates. Exposure of hippocampal slices to magnesium-free medium resulted in spontaneous epileptiform activity and a concurrent 38 +/- 5.47% decrease in CaM kinase II activity (range 38.8-75.4% of control; n = 7, P less than 0.001, paired Student's t test). The decrease in CaM kinase II activity was not reversible by treatment with protein phosphatases 1 and 2A (58.8 +/- 4.77% of control activity; range 28.6-69.7, P less than 0.01, paired Student's t-test), indicating that the decrease in CaM kinase II activity cannot be accounted for exclusively by autophosphorylation. The results demonstrate that magnesium-free medium treatment can induce spontaneous epileptiform activity and simultaneous changes in CaM kinase II activity.

Action Potentials

Regenerative, all-or-none electrographic seizures in the rat hippocampal slice in Mg-free and physiological medium.

All-or-none electrographic seizures (EGSs) were studied in hippocampal slices from young (21- to 38-day-old) rats in medium containing low (0 mM) or physiological (0.9 mM) levels of magnesium, with and without the GABAB agonist baclofen. Extracellular recording and stimulation were performed in stratum pyramidale and stratum radiatum of CA3, respectively. EGS activity was induced by exposure to low-Mg medium or by delivering repetitive stimulus trains in physiological Mg medium. After EGS activity had stabilized, the EGSs were tested for all-or-none behavior by varying the number of pulses in a train. An EGS was considered all-or-none if subthreshold stimulation produced no afterdischarge bursts, and if the EGS duration was largely independent of the number of suprathreshold stimulus pulses. According to this measure, EGSs in Mg-free + baclofen medium were all-or-none. EGSs evoked in physiological Mg medium were also all-or-none, although the threshold was higher, and the EGS duration lower, than in Mg-free medium. This all-or-none characteristic was observed whether the EGSs were induced by prior exposure to Mg-free medium or by repetitive stimulation, and in the presence and absence of baclofen. The all-or-none characteristic suggests that while the triggering mechanism for EGSs is strongly dependent on stimulus intensity, regenerative mechanisms--independent of stimulus intensity--are responsible for the maintenance of EGSs. EGSs are also terminated by mechanisms not dependent on stimulus intensity.

Action Potentials

DNA sequence evolution of the amylase multigene family in Drosophila pseudoobscura.

The alpha-Amylase locus in Drosophila pseudoobscura is a multigene family of one, two or three copies on the third chromosome. The nucleotide sequences of the three Amylase genes from a single chromosome of D. pseudoobscura are presented. The three Amylase genes differ at about 0.5% of their nucleotides. Each gene has a putative intron of 71 (Amy1) or 81 (Amy2 and Amy3) bp. In contrast, Drosophila melanogaster Amylase genes do not have an intron. The functional Amy1 gene of D. pseudoobscura differs from the Amy-p1 gene of D. melanogaster at an estimated 13.3% of the 1482 nucleotides in the coding region. The estimated rate of synonymous substitutions is 0.398 +/- 0.043, and the estimated rate of nonsynonymous substitutions is 0.068 +/- 0.008. From the sequence data we infer that Amy2 and Amy3 are more closely related to each other than either is to Amy1. From the pattern of nucleotide substitutions we reason that there is selection against synonymous substitutions within the Amy1 sequence; that there is selection against nonsynonymous substitutions within the Amy2 sequence, or that Amy2 has recently undergone a gene conversion with Amy1; and that Amy3 is nonfunctional and subject to random genetic drift.

Amylases

Amylase gene expression in intraspecific and interspecific somatic transformants of Drosophila.

The Amylase locus in Drosophila melanogaster normally contains two copies of the structural gene for alpha-amylase, a centromere-proximal copy, Amy-p, and a distal copy, Amy-d. Products of the two genes may display discrete electrophoretic mobilities, but many strains known to carry the Amy duplication are characterized by a single amylase electromorph, e.g., Oregon-R, which produces the mobility variant AMY-1. A transient expression assay was used in somatic transformation experiments to test the functional status of the Amy genes from an Oregon-R strain. Plasmid constructs containing either the proximal or distal copy were tested in amylase-null hosts. Both genes produced a functional AMY-1 isozyme. Constructs were tested against an AMY-3 reference activity produced by a coinjected plasmid that contains the Amy-d3 allele from a Canton-S strain. With reference to the internal control, the Amy-p and Amy-d genes from Oregon-R expressed different relative activity levels for AMY-1 in transient assays. The transient expression assay was successfully used to test the functional status of Amy-homologous sequences from strains of other species of Drosophila characterized by a single amylase elctromorph, namely, Drosophila pseudoobscura ST and Drosophila miranda S 204. The amylase-null strain of D. melanogaster provided the hosts for these interspecific somatic transformation experiments.

Amylases

NMDA antagonists differentiate epileptogenesis from seizure expression in an in vitro model.

In an electrographic model of seizures in the hippocampal slice, both of the N-methyl-D-aspartate (NMDA) antagonists 2-amino-5-phosphonovaleric acid and 5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate (MK-801) prevented the progressive development of seizures but did not block previously induced seizures. Thus, a process dependent on the NMDA receptor-ionophore complex establishes a long-lasting, seizure-prone state; thereafter the seizures depend on non-NMDA receptor-ionophore mechanisms. This suggests that there is an important distinction between epileptogenesis and seizure expression and between antiepileptogenic and anticonvulsant pharmacological agents.

2-Amino-5-phosphonovalerate

The drug MK-801 attenuates the development, but not the expression, of long-term potentiation and stimulus train-induced bursting in hippocampal slices.

Recent studies have demonstrated that (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801), a use-dependent blocker of N-methyl-D-aspartate (NMDA)-activated membrane channels, attenuates the development of long-term potentiation in vitro and kindling in vivo. Both of these phenomena are manifestations of physiological plasticity related to behavioural states and the results of these studies add to the gathering evidence for the involvement of the NMDA receptor/channel system in such processes. In the present experiment, slices of hippocampus, prepared from rats, were electrically stimulated to produce either long-term potentiation of the CA1 population spike or stimulus train-induced epileptiform bursting in area CA3. At 10 microM, MK-801 attenuated the development of long-term potentiation, but had no attenuating effect upon the previously-potentiated population spike. Similarly, 10 microM MK-801 attenuated the development of epileptiform activity in area CA3, but had little or no effect on the previously-established bursting in area CA3. These data support the suggestion that MK-801 exerts an antiepileptogenic, but not an anticonvulsant effect, at concentrations which also inhibit long-term potentiation.

Animals

Selection in natural and experimental populations of Drosophila pseudoobscura.

The inverted gene arrangements of Drosophila pseudoobscura were used by Th. Dobzhansky in pioneering analyses of natural selection. Recent experiments have shed light on the mechanisms of selection contributing to the balanced polymorphism for the gene arrangements. In experimental populations, both major components of fitness, viability and fertility, are frequency dependent, and rare genotypes often have a selective advantage. Viabilities are also density dependent. The frequency dependence and density dependence of the fitness components are not universal. Some karyotypes are strongly influenced by frequency or density, some are slightly influenced, and some do not appear to be influenced at all. The role of heterozygote advantage in the selection on the gene arrangements is not clear. It is probably one important element in the overall selection, but viability and fertility do not always show a heterozygote advantage. Viability and fertility components of selection seem to be about equally important in changing inversion frequencies. Male mating success is an important component of selection in natural populations, and in one population rare male karyotypes have been found to have a pronounced mating advantage.

Animals

Rare male mating advantage in a natural population of Drosophila pseudoobscura.

The natural selection acting on chromosomal inversions was studied in a natural population of Drosophila pseudoobscura. Females from this population were allowed to produce offspring from their matings in nature. They were then remated to males from a laboratory strain and again allowed to produce offspring. Offspring were also produced from matings of males from nature to laboratory females. Diagnosis of salivary chromosomes in these several sets of larval offspring allowed us to deduce the karyotypes of adult females and males from nature as well as the karyotypes of the offspring of these females by their matings in nature. We reason that the males collected with the females are a reasonable sample of those that mated the females and deposited the sperm they carried on capture. Chromosome frequencies in the offspring of wild females by their matings in nature were decomposed into male and female parental contributions. Changes in chromosome frequency due to male mating success were calculated by comparing chromosomal frequencies in adult males with those in the chromosomes they contributed to their offspring. These changes were sizable and provide direct evidence that male sexual selection is an important component of selection on the inversions in this natural population. We proceeded further to classify karyotypes on the basis of their frequencies and to calculate the fraction of offspring fathered by rare or common males. Rare male karyotypes as a group had a selective value nearly twice that of the common male karyotypes.

Animals