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At least 307 records · Page 17Linked to original sources

Application of three-dimensional molecular hydrophobicity potential to the analysis of spatial organization of membrane protein domains. II. Optimization of hydrophobic contacts in transmembrane hairpin structures of Na+, K(+)-ATPase.

A method of packing of transmembrane hairpin helices in proteins is described. The procedure is based on the optimization of hydrophobic contacts calculated using the three-dimensional (3D) molecular hydrophobicity potential technique. To verify the validity of the computational scheme, we calculated relative orientations of membrane-spanning peptides in pairs L2-L3, M2-M3, and M4-M5 from L- and M-subunits of the photoreaction center of Rhodopseudomonas viridis and compared the predicted structures with those derived from atomic coordinates. The results of computer modeling agree with the X-ray data. We applied the approach proposed to study possible interhelical interactions in transmembrane hairpin structures of Na+, K(+)-ATPase.

Cell Membrane↗

Application of three-dimensional molecular hydrophobicity potential to the analysis of spatial organization of membrane domains in proteins. III. Modeling of intramembrane moiety of Na+, K(+)-ATPase.

The most probable interlocation of transmembrane alpha-helices of Na+, K(+)-ATPase has been calculated by a computer-aided molecular simulation approach in the framework of models with eight and 10 helical peptides for the alpha-subunit. The method is based on the concept of three-dimensional molecular hydrophobicity potential (MHP) and provides valuable description of spatial hydrophobic properties of membrane-spanning segments as well as helix-helix packing interactions inside the membrane. Resulting model of the arrangement of intramembrane domain agrees with recent results on hydrophobic photolabeling of an intramembrane part of the beta-subunit and the sixth transmembrane segment of the alpha-subunit. It is also consistent with current ideas on hydrophobic organization of integral membrane proteins. Possible topology of a cation-binding site is discussed.

Cell Membrane↗

XY chromosome behaviour in the germ-line of the human male: a FISH analysis of spatial orientation, chromatin condensation and pairing.

We have used multicolour fluorescence in situ hybridization to study the behaviour of the X and Y chromosomes in relation to a representative autosome, chromosome 1, on air-dried testicular preparations from normal fertile human males. In a proportion of Sertoli cells at interphase as well as spermatogonial metaphases there is an apparent selective undercondensation of the heterochromatic block of the long arm of the Y, which may be of functional significance with respect to Y-specific gene activity, initiating and maintaining spermatogenesis; we suggest that this may involve a mechanism similar to heterochromatin position-effect variegation in Drosophila. In the supporting Sertoli as well as pre-meiotic and leptotene cells the X and Y occupy relatively restricted domains at opposite poles of the nuclear membrane, while the chromosome 1 centromere regions are located interstitially and appear prealigned. The XY pairing and 'sex vesicle' formation comprises a complex series of spatial movement and differential condensation patterns. On the basis of these observations we propose that: the XIST/Xist gene, known to be involved in somatic X inactivation, imposes a chromatin reorganization leading to bending at the X-inactivation centre both at first meiotic prophase in males and in the soma in females; and the differential X and Y segments are protected from potentially deleterious meiotic exchanges by their separate spatial orientation. In addition, there is an indication that the timing of pairing and first meiotic segregation of the sex chromosomes is different, and precocious in comparison to the pairing and segregation of the autosomes, which may explain the high incidence of sex chromosome aneuploidy in sperm.

Animals↗

Single motor unit analysis from spatially filtered surface electromyogram signals. Part I: spatial selectivity.

The aim of the study was to compare experimentally, on the basis of single motor unit (MU) activities, the selectivity of different spatial filters commonly used to detect surface electromyogram (EMG) signals. Surface EMG signals were recorded from the biceps brachii and the upper trapezius muscle of five subjects using a two-dimensional (2D) electrode array consisting of 16 pin electrodes. The subjects performed isometric contractions at different elbow angles and shoulder abduction and flexion. The same monopolar surface EMG signals were filtered using longitudinal single and double differential, transverse single and double differential and normal double differential filters. From the single MU action potentials, extracted by automatic EMG decomposition, indexes of transverse (perpendicular with respect to the fibre direction) and longitudinal (along the fibre direction) selectivity were computed. The number of detected MUs was 46 for the upper trapezius, with the arms held in the sagittal plane, and 52 when the arms were held in the frontal plane; 85 MUs were identified from the biceps brachii contractions. The results showed that transverse selectivity was significantly higher for the 2D and transverse one-dimensional (1D) filters with respect to the 1D longitudinal filters, whereas longitudinal selectivity was higher (i.e. MU action potentials were shorter) for the 2D filter and the longitudinal double differential filter. In particular, the relative attenuation of potential amplitude moving 5 mm from the source was, on average (for the two muscles), 16.5% for the least selective filter in the transverse direction (longitudinal single differential) and 35.7% for the most selective one in the same direction (transverse double differential). The MU action potential duration was, on average, 13.8 ms for the most selective filter in the longitudinal direction (longitudinal double differential) and 18.7 ms for the least selective one (transverse double differential). The normal double differential filter resulted in spatial selectivity indexes that ware not statistically different in the two directions from those of the best filters in each direction.

Action Potentials↗

Single motor unit analysis from spatially filtered surface electromyogram signals. Part 2: conduction velocity estimation.

The aim of the study was to compare experimentally conduction velocity (CV) estimates obtained with different estimation methods based on surface electromyogram (EMG) signals detected using five spatial filters. The filters investigated were the longitudinal single and double differential, transverse single and double differential, and normal double differential. The same surface EMG signals detected as described in Part 1 were used in this work. CV was estimated with four commonly used delay estimation techniques, i.e. from the distance between the peak values of two waveforms (with and without polynomial interpolation around the peak), and by the maximum likelihood estimate (MLE) based on two or more surface EMG channels. The average standard deviation of CV estimation (for all the MUs and the two muscles together) was 0.61 m s(-1) and 0.79 m s(-1) for the peak method, with and without interpolation, respectively, and 0.50 m s(-1) and 0.31 m s(-1) for the MLE method, from two and more surface EMG channels, respectively. Moreover, the mean of CV estimates varied by as much as 1 m s(-1) depending on the spatial filter used and the method adopted for CV estimation. Considering the dependence on the spatial filter only, the average (over all estimation methods) CV estimates obtained with the five spatial filters were 4.32 m s(-1) (normal double differential), 4.23 m s(-1) (longitudinal double differential), 4.61 m s(-1) (transverse double differential), 4.64 m s(-1) (transverse single differential) and 4.03 m s(-1) (longitudinal single differential). It was concluded that the comparison of single MU CV values obtained in different studies is critical if different spatial filters and processing techniques are used for their estimation. Higher estimates of CV were attributed to a smaller reduction in non-travelling signal components and thus were assumed to be positively biased.

Action Potentials↗

Functional analysis of spatially discriminative neurons in prefrontal cortex of rhesus monkey.

The present study addresses the question of whether prefrontal neurons that exhibit spatially selective patterns of discharge during the delay period in spatial delayed-response tasks code a mnemonic event. To examine this question, rhesus monkeys were trained to perform two variants of the classical spatial delayed-response task in both of which a delay intervened between cue presentation and response and the discriminative stimulus had to be recalled at the moment of response. They were also trained to perform two control tasks in which memory was not required since cues present throughout the delay informed the monkey of the correct response. Extracellular recordings were obtained from 192 neurons located in and around the principal sulcus of the frontal lobe during performance of both control and delay tasks. Comparison of the same neuron's activity across the 4 task conditions revealed a class of neuron that displayed spatially discriminative activity in the delay period only during delayed-response tasks and not during the same period of the control tasks. These neurons are candidates for units engaged in a central mnemonic process. Other neurons either exhibited similar activity in the delay period of control and delayed-response tasks or stronger discriminative behavior during this period in control tasks than in delayed response tasks. We conclude that delay-related spatially discriminative neurons found in the prefrontal association cortex are diversified and that certain of them play a specific role in mnemonic coding.

Animals↗

An analysis of spatial navigation in prenatally protein malnourished rats.

Developing rats were either malnourished or adequately nourished during the prenatal period by feeding their dams diets of low (6% casein) or adequate (25% casein) protein content 5 weeks prior to mating and throughout pregnancy. All pups received adequate nutrition from the day of birth onwards. Male offspring were tested in one of two spatial navigation tests in the Morris water tank. In proximal-cue tests (postnatal days 16-20), the position of a platform, which provided a means to escape from swimming, was denoted by an obvious visual cue located directly on the platform. In distal-cue tests (postnatal days 20-27 and adult ages, days 70-71 and days 220-221), the escape platform was submerged below the surface of the water so that the rats were required to use extramaze visual cues to guide them to the platform. Neither proximal-cue nor distal-cue navigation was significantly impaired in the prenatally malnourished rats relative to controls, at any of the ages tested.

Animals↗

A choice reaction time analysis of spatial frequency discrimination.

Simple reaction time to the onset of sinewave gratings was measured as a function of spatial frequency in two observers. These results are compared to the choice reaction time required for the observer to correctly discriminate the spatial frequency of two gratings flashed sequentially. Grating contrast was either 0.75 or 1.5 logarithmic units above the detection threshold for each spatial frequency tested. The spatial phase and contrast of the reference and test gratings were varied from trial to trial by small random amounts to eliminate fixed cues other than the difference frequency. The spatial frequency difference between the reference and test grating was either 0.125, 0.25 or 0.5 octave. As has been earlier reported, simple reaction time increases with increasing spatial frequency. Contrary to this, choice reaction time first increases (up to 4 c/deg) and then decreases. We derived the time required by the observer to make a spatial frequency judgment by subtracting the simple reaction time from the choice reaction time for a given spatial frequency and contrast. The maximum decision time occurs in the medium spatial frequency range (between 1 and 4 c/deg), at which frequencies we are most sensitive. The time required to make a correct spatial-frequency discrimination decreases with increasing spatial-frequency difference. The decision time is, however, fairly invariant over a large range of suprathreshold contrast levels. The findings suggest that the decision time for spatial frequency discrimination increases with the number of mechanisms involved.

Contrast Sensitivity↗

Nonlinear analysis of spatial vision using first-and-second-order Volterra transfer functions measurement.

The harmonic input method of nonlinear system identification is modified to allow the Volterra series approach to be used for psychophysical investigation of various aspects of human pattern vision in the spatial frequency domain. While it is well known that only one modulation transfer function provides a complete characterization of a linear system, a number of multidimensional transfer functions are needed to identify a nonlinear system. We have shown, that so far as the contrast sensitivity to sine-wave gratings may be used for an empirical estimate of the first-order modulation transfer function of the human visual system, the contrast sensitivity to difference harmonics may be used as an empirical estimate of the second-order modulation transfer function. A difference harmonic arises from a mixture of two sine-wave gratings resulting from the nonlinearity of the visual system. Difference harmonic, experienced as some periodic beatlike structure, may still be observed if frequencies of the component gratings are higher than the maximum visual acuity. The visibility of the low-frequency beatlike pattern produced by pairs of sine-wave gratings, which themselves are of spatial frequencies too high to be resolved, could be accounted for either by a difference frequency distortion product (Burton, 1973) or by a special beat detector (Derrington & Badcock, 1985). We found that increasing the contrast of one component grating may be compensated for by reducing the contrast of the other component grating, the beatlike pattern being at threshold. This is exactly what would be expected if the beatlike pattern is detected because of the difference harmonics produced by nonlinearity of the visual system. We have determined contrast thresholds for the difference harmonics which occur between two unresolved different spatial frequencies. The contrast sensitivity function for difference harmonics was found to have a marked similarity both in the shape and position of peak sensitivity to the contrast sensitivity function for single sine-wave gratings. Another important characteristic of the contrast sensitivity function for difference harmonics is that it depends only on the frequency difference, delta f = f1 - f2, rather than on the value of either f1 or f2. All this indicates that a difference harmonic arises from local nonlinearities in the visual system. More specifically, the visual system may be represented as a cascade system, composed of a linear system with transfer function O (f) followed by a nonlinear element, r(.), without spatial spread in cascade with another linear system with transfer function P (f).(ABSTRACT TRUNCATED AT 400 WORDS)

Contrast Sensitivity↗

A behavioural analysis of spatial localization following electrolytic, kainate- or colchicine-induced damage to the hippocampal formation in the rat.

This experiment examines the notion that in the rat the hippocampal formation is an essential structure in the neurological representation of spatial abilities. Spatial localization by rats with different types of hippocampal damage, including bilateral electrolytic lesions, unilateral and bilateral kainic acid-induced CA3-CA4 lesions, and unilateral and bilateral colchicine-induced dentate gyrus lesions, was compared with vehicle-injected and normal control groups in the Morris water task. The task required the rats to escape from cold water by finding a submerged and hidden platform located at a fixed place within the room. The start point was varied randomly from trial to trial and there were no local cues available to indicate the position of the hidden platform. After training, the platform was moved. Escape latencies and the initial swimming headings revealed that all lesion groups, except the unilateral CA3-damaged group, were impaired at finding the platform: the dentate-damaged rats exhibited the greatest deficit. When the platform was moved the control rats swam mainly in the part of the pool that had previously contained the platform and, on finding it in the new location, they showed a marked dishabituation of rearing. None of the bilateral lesion groups showed these effects.

Animals↗

Confocal image analysis of spatial variations in immunocytochemically identified calmodulin during pollen hydration, germination and pollen tube tip growth in Nicotiana tabacum L.

Using monoclonal anti-calmodulin antibodies in conjunction with confocal scanning laser microscopy we have analysed the spatial variations in the distribution pattern of calmodulin (CaM) during the sequential events of pollen hydration, germination and tube growth in Nicotiana tabacum. These immunocytochemical observations have been complemented by immunochemical studies wherein the anti-calmodulin antibody raised against pea CaM recognises a polypeptide of c. 18 kDa in the pollen extracts. Digitisation of confocally acquired optical sections of immunofluorescence images reveals that in hydrated pollen a high level of CaM is consistently present in the region of the germinal apertures. Subsequently, with the onset of germination a high CaM concentration was found associated with the plasma membrane of the germination bubble and in the cytoplasm in its vicinity, while in the vegetative cytoplasm a weak diffuse and intense punctate signal was registered. CaM immunostain was also detected in association with the plasma membrane of the tube tips in both short and long pollen tubes. Furthermore, the cytosol of the tubes invariably manifested an apically focused CaM gradient. We were, however, unable to detect any vacuolar association of CaM in the older regions of the pollen tubes. Although punctate immunostain was obvious across the pollen tube numerous punctate structures were invariably present in the extreme tip. The possible implications of these findings in development of cell polarity, polarised growth, maintenance of calcium homeostasis and CaM interactions with other mechanochemical motor proteins in effecting propulsion of organelles during pollen hydration, germination and pollen tube growth are discussed.

Blotting, Western↗