Arithmetic of subthreshold synaptic summation in a model CA1 pyramidal cell

The rules of synaptic integration in pyramidal cells remain obscure, in part due to conflicting interpretations of existing experimental data. To clarify issues, we developed a CA1 pyramidal cell model calibrated with a broad spectrum of in vitro data. Using simultaneous dendritic and somatic recordings and combining results for two different response measures (peak versus mean EPSP), two different stimulus formats (single shock versus 50 Hz trains), and two different spatial integration conditions (within versus between-branch summation), we found that the cell's subthreshold responses to paired inputs are best described as a sum of nonlinear subunit responses, where the subunits correspond to different dendritic branches. In addition to suggesting a new type of experiment and providing testable predictions, our model shows how conclusions regarding synaptic arithmetic can be influenced by an array of seemingly innocuous experimental design choices.     

Heart specific genes revealed by EST sampling

Background  

Cardio-vascular diseases are the first cause of death worldwide, particularly in the developed countries; the identification of genes specifically expressed in the cardiac muscle is thus of major biomedical interest. In this study, we performed a comprehensive analysis of the expression profiles to identify genes over-expressed in the human adult heart using the public Expressed Sequence Tags (ESTs) database. The initial set of genes expressed in the heart was constructed by clustering and assembling ESTs from human adult heart cDNA libraries. Expression profiles were then generated for each of these genes by counting their cognate ESTs in all libraries. Differential expression was assessed by applying to these profiles a previously published statistical procedure.     

Kinesin-mediated organelle translocation revealed by specific cellular manipulations

The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3'- dihexyloxacarbocyanine iodide (DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, and a parallel accumulation of label within the cell center. On the other hand, mitochondria microtubules and microfilaments display a distribution that closely resembles that observed in control cells. KHC suppression of neurons and astrocytes completely inhibited the Brefeldin A-induced spreading and tubulation of the Golgi- associated structure enriched in mannose-6-phosphate receptors. In addition, KHC suppression prevents the low pH-induced anterograde redistribution of late endocytic structures. Taken collectively, these observations suggest that in living neurons, kinesin mediates the anterograde transport of tubulovesicular structures originated in the central vacuolar system (e.g., the endoplasmic reticulum) and that the regulation of kinesin-membrane interactions may be of key importance for determining the intracellular distribution of selected organelles.     

Intergenomic conflict revealed by patterns of sex-biased gene expression

Intergenomic conflict can affect the distribution of genes across eukaryotic genomes. Because the phenotypic optima of males and females often differ, the fitness consequences of newly arisen alleles might not be concordant between the sexes and can be sexually antagonistic--genetic variants favored in one sex are deleterious in the other. In this article, we demonstrate that previously unexplained patterns of sex-biased gene expression in Drosophila melanogaster might have evolved by sexual antagonism, and that the majority of sex-biased expression is due to adaptive changes in males, implying that males experience stronger selection than females.     

Differential transgene expression patterns in Alzheimer mouse models revealed by novel human amyloid precursor protein‐specific antibodies

Alzheimer's disease (AD) is histopathologically characterized by neurodegeneration, the formation of intracellular neurofibrillary tangles and extracellular Aβ deposits that derive from proteolytic processing of the amyloid precursor protein (APP). As rodents do not normally develop Aβ pathology, various transgenic animal models of AD were designed to overexpress human APP with mutations favouring its amyloidogenic processing. However, these mouse models display tremendous differences in the spatial and temporal appearance of Aβ deposits, synaptic dysfunction, neurodegeneration and the manifestation of learning deficits which may be caused by age‐related and brain region‐specific differences in APP transgene levels. Consequentially, a comparative temporal and regional analysis of the pathological effects of Aβ in mouse brains is difficult complicating the validation of therapeutic AD treatment strategies in different mouse models. To date, no antibodies are available that properly discriminate endogenous rodent and transgenic human APP in brains of APP‐transgenic animals. Here, we developed and characterized rat monoclonal antibodies by immunohistochemistry and Western blot that detect human but not murine APP in brains of three APP‐transgenic mouse and one APP‐transgenic rat model. We observed remarkable differences in expression levels and brain region‐specific expression of human APP among the investigated transgenic mouse lines. This may explain the differences between APP‐transgenic models mentioned above. Furthermore, we provide compelling evidence that our new antibodies specifically detect endogenous human APP in immunocytochemistry, FACS and immunoprecipitation. Hence, we propose these antibodies as standard tool for monitoring expression of endogenous or transfected APP in human cells and APP expression in transgenic animals.     

Precedence of spatial pattern learning revealed by immediate reversal performance

Human participants learned to choose eight correct locations in a 4 × 4 matrix on a computer display. The locations were arranged either in a structured spatial pattern or an unstructured but consistent spatial arrangement. When the assignment of correct and incorrect locations was reversed after initial learning, participants in the spatial pattern condition demonstrated reversal performance immediately (i.e., following the first choice after reversal of the contingencies). Follow-up experiments confirmed that immediate reversal performance depends on a structured spatial pattern among the locations and that a learned motor pattern cannot explain the immediate reversal performance. This pattern of results shows that learning the spatial relations among locations has precedence over learning about the individual locations, even when the individual locations are completely valid predictive cues.     

Detection and pattern of interspecific hybridization between Gliricidia sepium and G. maculata in Meso-America revealed by PCR-based assays

Gliricidia sepium provides a variety of products important for rural communities in tropical countries. Native populations in Meso-America currently form an important source of seed for distribution to farmers, but concerns centre on mechanisms which may lead to their genetic erosion, including anthropogenic dispersal and subsequent introgression from the related species, G. maculata. Populations of Gliricidia were examined genetically using approaches based on the polymerase chain reaction to test for interspecific hybridization and introgression between G. sepium and G. maculata. Analysis involved 13 RAPD and two RFLP-PCR markers which were identified to have species-diagnostic distributions. Data from both approaches corresponded and indicated three locations where multilocus genotypes were consistent with an hybrid origin. Data at one of these sites was consistent with introgression following hybridization. The hybrid origin of populations was supported by the intermediate geographical location of these sites to ‘pure’ populations of each species. Analysis of maternally inherited organellar DNA, which involved the detection of SSCPs in mitochondrial DNA amplification products, allowed further delineation of genetic structure among Gliricidia populations. Mitochondrial data indicated a high degree of organelle differentiation between sampled locations and identified G. sepium- and G. maculata-diagnostic haplotypes. This data supported the interpretation of genetic structure based on RAPDs and RFLP-PCR. In addition, cytonuclear analysis allowed the directionality of gene transfer during the formation of hybrid populations to be described. Despite evidence for the occurrence of interspecific hybridization and introgression in Gliricidia, important resource populations of G. sepium on the Pacific coast appear to have retained their genetic integrity. Implications in terms of the conservation and utilization of genetic resources within the genus are discussed.     

Adhesion patterns of cell interactions revealed by reflection contrast microscopy

Reflection contrast in combination with phase contrast microscopy was utilized for the study of adhesion patterns of locomotive L5222 rat leukemia cells. It was found that for cells moving in a spherical shape on the glass surface, adhesions were very faint. This inconspicuous pattern, however, became very distinct, as soon as the cells changed to a flattened configuration. Such a change took place when leukemia cells came into contact with other spread cells and started to move under these cells. Reflection contrast further showed that in the pathway of the locomoting L5222 cells the adhesions of the overlying spread cells were momentarily detached from the substrate. It is concluded that the combination of reflection contrast and phase contrast represents a good tool for gaining new information on the interaction of motility and formation of adhesions.     

Banding patterns of Chinese hamster chromosomes revealed by new techniques

Two kinds of techniques were newly developed to reveal banding patterns of the Chinese hamster chromosomes. Both techniques were essentially the same as those used for the extraction of proteins, and one of them could induce bands in chromosome arms in only a few seconds. Banding patterns produced by these techniques appeared to be identical to those induced by the methods reported by previous workers, requiring post-fixation incubation of slides in a warm saline. —The banding pattern was typical for each chromosome pair, permitting unequivocal identification of several pairs which were hardly distinguished by the conventional staining procedures. It was confirmed that these patterns had been well preserved in the chromosomes of a cultured cell line.Contribution No. 870 from the National Institute of Genetics, Japan. This work was supported in part by a grant (No. 9001) from the Ministry of Education in Japan.     

Temporal patterns recognized by a network of coordinated time delays and coincidence detectors

A computational model of a neuronal network is described which performs a fundamental task of general perception: recognition of temporal patterns in continuous and uncued neuronal spike trains. The presented network is able to recognize each pattern element (100 ms interval composed of sets of 10, 20, 30 and 40 ms interspike intervals combined in linear order) as it arrives. Its operation is based upon biologically plausible filtering mechanisms and population neurodynamics.     

Dependence of EPSP summation pattern on membrane potential in mollusk neurons

The dependence of monosynaptic EPSP pattern on membrane potential level was investigated during experiments on identifiedPlanorbis corneus neurons. A random sequence of twin stimuli spaced at different intervals was used to ascertain this pattern. It was shown that the value of interstimulus interval corresponding to the most marked EPSP interactions mainly depended on postsynaptic mechanisms: selected changes in the parameters of the postsynaptic neuron leads to similar alteration in the summation function extremum. The possibility arises from these results of selective tuning (applying to each input or groups of inputs) to specific intervals (or frequencies); this tuning may be modulated according to changing membrane potential at the relevant portion of the neuron. The possible functional role of this mechanism is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 3, pp. 314–320, May–June, 1989.     

Cytoplasmic actin patterns in Physarum as revealed by NBD-phallacidin staining

Fluorescently labeled phallacidin, a F-actin specific drug, was used to demonstrate the morphological variety in the cytoskeletal actin pattern of thin-spread plasmodia of the acellular slime mould Physarum polycephalum. The patterns observed in phallacidin-stained specimens consisted of a polygonal network in the anterior region, and of longitudinal as well as helically twisted fibrils in plasmodial strands of the posterior region. These observations are in complete accordance with our recent results obtained on comparable plasmodia by immunofluorescence microscopy using specific antibodies against actin.     

Spatial patterns of plant diversity below-ground as revealed by DNA barcoding

Our understanding of the spatial organization of root diversity in plant communities and of the mechanisms of community assembly has been limited by our ability to identify plants based on root tissue, especially in diverse communities. Here, we test the effectiveness of the plastid gene rbcL, a core plant DNA barcoding marker, for investigating spatial patterns of root diversity, and relate observed patterns to above-ground community structure. We collected 3800 root fragments from four randomly positioned, 1-m-deep soil profiles (two vertical transects per plot), located in an old-field community in southern Ontario, Canada, and extracted and sequenced DNA from 1531 subsampled fragments. We identified species by comparing sequences with a DNA barcode reference library developed previously for the local flora. Nearly 85% of sampled root fragments were successfully sequenced and identified as belonging to 29 plant species or species groups. Root abundance and species richness varied in horizontal space and were negatively correlated with soil depth. The relative abundance of taxa below-ground was correlated with their frequency above-ground (r = 0.73, P = 0.0001), but several species detected in root tissue were not observed in above-ground quadrats. Multivariate analyses indicated that diversity was highly structured below-ground, and associated with depth, root morphology, soil chemistry and soil texture, whereas little structure was evident above-ground. Furthermore, analyses of species co-occurrence indicates strong species segregation overall but random co-occurrence among confamilials. Our results provide insights into the role of environmental filtering and competitive interactions in the organization of plant diversity below-ground, and also demonstrate the utility of barcoding for the identification of plant roots.     

Dietary obesity caused by a specific circadian eating pattern

The eating pattern is altered by high-fat diet-induced obesity. To clarify whether this is dependent on the fatty acid profile of the diet, the authors conducted two studies on adult female Sprague-Dawley rats fed normal-fat chow or high-fat diets with varying fatty acid composition. Eating pattern and body weight were assessed in rats fed canola-based (low in saturated fatty acids) or lard-based (moderate in saturated fatty acids) diets for 7 days, and in animals fed chow or canola- or butter-based diets (rich in saturated fatty acids) for 43 days. These parameters were also determined when restricted amounts of low-fat canola- or butter-based diets were consumed for 25 days. Early exposure to canola or lard high-fat feeding or prolonged access to canola- or butter-based fat-rich diets (relative to chow feeding) did not alter the normal light-dark distribution of food and energy intake. All animals ingested most of their food during the dark phase. However, feeding the high-fat canola- and butter-based diets produced an altered eating pattern during the light phase characterized by a smaller number of meals, longer intermeal interval, and enhanced satiety ratio, and consumption of shorter-lasting meals than chow-fed animals. Relative to canola or chow feeding, butter-fed animals consumed a lower number of meals during the dark phase and had a higher eating rate in the light phase, but ate larger meals overall. Only butter feeding led to overeating and obesity. When given a restricted amount of low-fat canola- or butter-based diet at the start of the light phase, rats ate most of their food in that phase and diurnal rather than nocturnal feeding occurred with restriction. These findings underscore the role of saturated fatty acids and the resulting eating pattern alteration in the development of obesity.     

Features of lineage‐specific hematopoietic metabolism revealed by mitochondrial proteomics

Hematopoietic bone marrow is a regenerative tissue of high clinical relevance, yet relatively little is known about the metabolism of the stem and progenitor populations concerned. We have used a multipotent murine cell line to generate sufficient numbers of cells undergoing self‐renewal, erythroid or myeloid differentiation to allow a proteomics analysis of enriched mitochondria. Stringent analysis identified 37 mitochondria‐associated proteins changing on differentiation in this system. Those induced during differentiation were commonly associated with mature cell functions, while those inactivated upon differentiation indicate widespread changes in mitochondrial transport, fatty acid catabolism and oxidative phosphorylation. An erythroid specific reduction in glutamate pyruvate amino transferase 2 was confirmed at the protein level by Western blotting and at the functional level by assays of metabolite turnover. In addition to validating the dataset, these findings suggest significant differences in the core‐metabolism between erythropoiesis and myelopoiesis. This knowledge is of relevance to the in vitro production of cell therapy products and to studies of the interdependence of metabolic and signaling pathways in regenerative tissues. Data are available via ProteomeXchange with identifier PXD002968.