An emergent mechanism of selective visual attention in Drosophila

Due to the limited computational capacity of visual systems and the limited capacity to perform several mental operations at once, animals only select a small proportion of the stimuli available at any one time. It remains to be clarified how this process is related to the spatio-temporal dynamics of cell assemblies in the brain. By employing the flight simulator, selective visual attention behavior is studied in Drosophila. It has been found that for the visual objects presented, the tethered fruitflies display various attention patterns. Specifically, the learning memory mutants dunce and amnesiac possess attention patterns totally different from that of the wild-type fly. To explain these results from the viewpoint of dynamic cell assemblies, a neural network has been developed in which a possible link between the activity of cell assemblies, encoding of sensory information, and selective attention in Drosophila is proposed. Received: 3 November 1998 / Accepted in revised form: 1 July 1999     

An automated climbing apparatus to measure chemotherapy-induced neurotoxicity in Drosophila melanogaster

We have developed a novel model system in Drosophila melanogaster to study chemotherapy-induced neurotoxicity in adult flies. Neurological deficits were measured using a manual geotactic climbing assay. The manual assay is commonly used; however, it is laborious, time-consuming, subject to human error and limited to observing one sample at a time. We have designed and built a new automated fly-counting apparatus that uses a “video capture-particle counting technology” to automatically measure 10 samples at a time, with 20 flies per sample. Climbing behavior was assessed manually, as in our previous studies, and with the automated apparatus within the same experiment yielding statistically similar results. Both climbing endpoints as well as the climbing rate can be measured in the apparatus, giving the assay more versatility than the manual assay. Automation of our climbing assay reduces variability, increases productivity and enables high throughput drug screens for neurotoxicity.     

A new paradigm for operant conditioning of Drosophila melanogaster

A freely walking single fly (Drosophila melanogaster) can be conditioned to avoid one side of a small test chamber if the chamber is heated whenever the fly enters this side. In a subsequent memory test without heat it keeps avoiding the heat-associated side. The memory mutants dunce ¹ and rutabaga ¹ successfully avoid the heated side but show no avoidance in the memory test. Wildtype flies can be trained to successively avoid alternating sides in a reversal conditioning experiment. Every single fly shows strong avoidance and a positive memory score. The new conditioning apparatus has several advantages: (1) Statistically significant learning scores can be obtained for individual flies. (2) Learning scores are obtained fully automatically without interference of the experimenter. (3) The procedure is fast, robust and requires little handling. Therefore the apparatus is suitable for largescale mutant screening. (4) Animals are not attached to a hook and thus can easily be used for breeding.Abbreviations dnc dunce gene - PI performance Index - rut rutabaga gene - S.E.M. standard error of mean     

An application of gustatory psychophysics in Antarctica

Measures of gustatory perception were made on a team of ninemen who wintered-over in Antarctica between December 1976 andOctober 1977, at Scott Base, Ross Dependency. Contrary to expectations,no decrement in performance was found, but some individual stabilityof response patterns and on average a slight improvement inacuity was recorded. ¹ This study was conducted under the administration of the NewZealand Department of Scientific and Industrial Research AntarcticDivision, supported by a grant from the Medical Research Councilof New Zealand, and manned by personnel from the Universityof Canterbury and Victoria University of Wellington. It formedpart of the program coded as Event 17, 1976–77.     

Concurrent TMS-fMRI and psychophysics reveal frontal influences on human retinotopic visual cortex

BACKGROUND: Regions in human frontal cortex may have modulatory top-down influences on retinotopic visual cortex, but to date neuroimaging methods have only been able to provide indirect evidence for such functional interactions between remote but interconnected brain regions. Here we combined transcranial magnetic stimulation (TMS) with concurrent functional magnetic resonance imaging (fMRI), plus psychophysics, to show that stimulation of the right human frontal eye-field (FEF) produced a characteristic topographic pattern of activity changes in retinotopic visual areas V1-V4, with functional consequences for visual perception. RESULTS: FEF TMS led to activity increases for retinotopic representations of the peripheral visual field, but to activity decreases for the central field, in areas V1-V4. These frontal influences on visual cortex occurred in a top-down manner, independently of visual input. TMS of a control site (vertex) did not elicit such visual modulations, and saccades, blinks, or pupil dilation could not account for our results. Finally, the effects of FEF TMS on activity in retinotopic visual cortex led to a behavioral prediction that we confirmed psychophysically by showing that TMS of the frontal site (again compared with vertex) enhanced perceived contrast for peripheral relative to central visual stimuli. CONCLUSIONS: Our results provide causal evidence that circuits originating in the human FEF can modulate activity in retinotopic visual cortex, in a manner that differentiates the central and peripheral visual field, with functional consequences for perception. More generally, our study illustrates how the new approach of concurrent TMS-fMRI can now reveal causal interactions between remote but interconnected areas of the human brain.     

Arthropod segmentation: beyond the Drosophila paradigm

Most of our knowledge about the mechanisms of segmentation in arthropods comes from work on Drosophila melanogaster. In recent years it has become clear that this mechanism is far from universal, and different arthropod groups have distinct modes of segmentation that operate through divergent genetic mechanisms. We review recent data from a range of arthropods, identifying which features of the D. melanogaster segmentation cascade are present in the different groups, and discuss the evolutionary implications of their conserved and divergent aspects. A model is emerging, although slowly, for the way that arthropod segmentation mechanisms have evolved.     

An alternative paradigm for hominoid evolution

Biological clock data from protein sequences indicate times for the divergence of humans and African apes that are only 2–3 million years before the present and hence inconsistent with the generally accepted view ofAustralopithecus on the evolutionary line toHomo. This inconsistency has been reconciled for most investigators by postulating a slowing of the biological clock among higher primates. However, there is no independent evidence for such a slowing, and for a number of reasons a specific slowing is unlikely. Therefore, an alternative paradigm is considered here based on the hypothesis that the molecular clock data are correct. The main consequence of this hypothesis is the placement ofA. afarensis in a position ancestral to African apes. An argument in support of this alternative paradigm is formulated concerning the evolution of knuckle-walking in African apes from ancestors whose bipedalism was already well developed. Published data are cited, particularly concerning the structure of the wrist, which accommodate poorly the evolution of African apes from palmigrad-walking or brachiating ancestors resemblingProconsul africanus orPongo. These arguments suggest that an alternative paradigm of hominoid evolution placingA. afarensis ancestral toHomo, Gorilla, andPan warrants further consideration.     

An automated method for acetylcholinesterase kinetics

An automated assay for acetylcholinesterase (EC 3.1.1.7.) has been developed based on the manual spectrophotometric method of Ellmanet al. (1). This method was used to determine (a) the enzyme activity of an unknown sample and (b) the dependence of initial rates given by a fixed enzyme concentration on the substrate concentration. Methods to minimize possible enzyme modification by DTNB (2) are described. Finally a modification of the conventional autoanalyser procedure permitted rapid and reproducible enzyme kinetic analysis under various conditions. This helped to minimize the effects of possible enzyme inactivation at high dilutions especially when using crude enzyme preparations.     

An automated coordinate recorder for biometry

Many measuring devices used in micropaleontology are unsatisfactory because observations must be recorded manually. Moreover, most use a linear scale that depends on the presence of predetermined morphological loci. An x, y recorder, largely adapted from instruments available in geological laboratories, is discussed. It obviates manual recording, frees data collection from the constraint of homologous loci and. in some applications, permits more shape information to be collected.     

An automated assay for phosphorylase kinase

A fully automated assay of phosphorylase kinase capable of processing 40 samples/hr is described. Problems due to enzyme adsorbed to the incubation coil and thereby producing a continuous rise of the base line were overcome by washing with a solution containing 0.05% Triton X-100, 0.03% SDS, and 5 mm EDTA. Under these conditions, a linear relationship between phosphorylase kinase concentration in the incubation mixture and absorbancy at pH's 6.8 and 8.2 is obtained when 1% glycogen is present in the reaction mixture.Inclusion of glycogen allows reduction of the concentration of phosphorylase in the incubation mixture. Due to the presence of Triton X-100 in this test, the carbohydrate acts like an allosteric activator of phosphorylase kinase. The standard deviation of the automated kinase test is 1.3% lower than that of the manual test.     

An automated assay for brain serotonin

This communication describes an automated assay for brain serotonin. The assay is an adaptation of a commonly used manual assay which utilizes the reaction of o-phthalaldehyde with serotonin to develop fluorescence (2). In the automated assay the samples to be analyzed were mixed with 7.5 N HCl and o-phthalaldehyde in the presence of reduced glutathione, heated at 80°C, cooled and their fluorescence recorded. Linearity with respect to serotonin was demonstrated from 0 to at least 5 μM, and the lower limit of sensitivity was 0.7 ng serotonin (0.4 ml of 0.01 μM). The % error (standard deviation times 100 divided by the mean) was always 2% or less. The specificity was studied, and the assay was applied to the measurement of rat brain serotonin levels by demonstrating an increase in brain serotonin levels after pargyline treatment and a decrease after reserpine treatment.     

Drosophila mutants disturbed in visual orientation

With the Y-maze selection technique described in Part I 2 strains of probably central nervous system (CNS) mutants have been isolated. These mutants show defects in the computation of both position and movement. One of these mutants (nofE^B12) shows strong avoidance of small patterns moving with high velocity. This inversion of the object-induced orientation response can be mainly attributed to a modification of responses to fast progressive (front-to-back) movement. It is thus possible that overall optomotor behaviour may be decomposed into a set of genetically independent modules.     

Blue adaptation: An experimental tool for the study of visual receptor mechanisms and behaviour of Drosophila

Physiological and behavioural studies with Drosophila to elucidate visual mechanisms have exploited the bi-stability of the visual pigment in the peripheral retinula cells R1–6, and the off-on switch action of blue and orange light. Measurements of flicker fusion and response waveform from both receptor and lamina regions prior and subsequent to blue adaptation, which induces a prolonged depolarising afterpotential and loss of visual function in R1–6, show these retinula cells to have a high fusion frequency and R7/8, the central retinula cells, a lower fusion frequency. Such measurements also allow analysis of the extracellular response in terms of contributing cells, and its potential for studying the fly's ability to respond to various potential visual cues such as a rotating plane of polarised light. Blue adapted flies fail to fixate normally a black stripe, confirming a role for R1–6 in orientation behaviour requiring a competent degree of acuity.Based on material presented at the European Neurosciences Meeting, Florence, September 1978     

Drosophila wingless: A paradigm for the function and mechanism of Wnt signaling

The link between oncogenesis and normal development is well illustrated by the study of the Wnt family of proteins. The first Wnt gene (int-1) was identified over a decade ago as a proto-oncogene, activated in response to proviral insertion of a mouse mammary tumor virus. Subsequently, the discovery that Drosophila wingless, a developmentally important gene, is homologous to int-1 supported the notion that int-1 may have a role in normal development. In the last few years it has been recognized that int-1 and Wingless belong to a large family of related glyco-proteins found in vertebrates and invertebrates. In recognition of this, members of this family have been renamed Wnts, an amalgam of int and Wingless. Investigation of Wnt genes in Xenopus and mouse indicates that Wnts have a role in cell proliferation, differentiation and body axis formation. Further analysis in Drosophila has revealed that Wingless function is required in several developmental processes in the embryo and imaginal discs. In addition, a genetic approach has identified some of the molecules required for the transmission and reception of the Wingless signal. We will review recent data which have contributed to our growing understanding of the function and mechanism of Drosophila Wingless signaling in cell fate determination, growth and specification of pattern.     

An automated system for karyotyping mouse chromosomes

A system developed for interactive automated analysis of human chromosomes has been modified for use with mouse cells. The system is described, with emphasis on those features that facilitate retaining for different preparations or even different species. Some performance figures are presented.     

An automated, handheld biosensor for aflatoxin

A new immunoaffinity fluorometric biosensor has been developed for detecting and quantifying aflatoxins, a family of potent fungi-produced carcinogens that are commonly found in a variety of agriculture products. They have also been cited as a biological agent under weapons development. The handheld, self-contained biosensor is fully automatic, highly sensitive, quick, quantitative, and requires no special storage. Approximately 100 measurements can be made before refurbishment is required, and concentrations from 0.1 parts per billion (ppb) to 50 ppb can be determined in <2 min with a 1 ml sample volume. The device operates on the principles of immunoaffinity for specificity and fluorescence for a quantitative assay. The analytic procedure is flexible so that other chemical and biological analytes could be detected with minor modifications to the current device. Advances in electro-optical components, electronics, and miniaturized fluidics were combined to produce this reliable, small, and versatile instrument.     

An efficient paradigm for genetic epidemiology cohort creation

Development of novel methodologies to efficiently create large genetic epidemiology cohorts is needed. Here we describe a rapid, precise and cost-efficient method for collection of DNA from cases previously experiencing an osteoporotic fracture by identifying cases using and administrative health-care databases. Over the course of 14 months we collected DNA from 1,130 women experiencing an osteoporotic fracture, at a cost of $54 per sample. This cohort is among the larger DNA osteoporotic fracture collections in the world. The novel method described addresses a major unmet health care research need and is widely applicable to any disease that can be identified accurately through administrative data.     

An algorithm for automated closure during assembly

Background  

Finishing is the process of improving the quality and utility of draft genome sequences generated by shotgun sequencing and computational assembly. Finishing can involve targeted sequencing. Finishing reads may be incorporated by manual or automated means. One automated method uses targeted addition by local re-assembly of gap regions. An obvious alternative uses de novo assembly of all the reads.