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PubMed1994

Biochemical evidence for the orientation of cytochrome b in the yeast mitochondrial membrane in the eight-helix model.

The topographical localization of the N-terminus of cytochrome b in the inner mitochondrial membrane was determined by mild proteolysis of the yeast mitochondrial cytochrome bc1 complex and identification of the proteolytic fragments derived from subunits of the complex with an established orientation in the inner membrane. The cytochrome bc1 complex was incorporated into proteoliposomes which were separated by cytochrome c affinity chromatography into two populations in either the mitochondrial or the submitochondrial orientation. Core protein I which protrudes from the matrix side of the inner membrane was digested by proteinase K only in proteoliposomes with the submitochondrial orientation and not in those with the mitochondrial orientation. By contrast, cytochrome c1 with protrudes from the cytoplasmic side of the inner membrane was digested by proteinase K only in proteoliposomes with the mitochondrial orientation and not in those with the submitochondrial orientation. Cytochrome b was digested by SV8 protease only in proteoliposomes with the mitochondrial orientation to yield two aggregating fragments of 25.6 and 24.5 kDa. These peptides were isolated by preparative gel chromatography and sequenced to establish that the cleavage of cytochrome b by SV8 protease occurred at glutamate residues 59 and 66. These residues are localized in the extramembranous loop between the two hydrophobic membrane-spanning helices A and B and thus face the cytoplasmic side of the inner mitochondrial membrane. These results indicate that the N-terminus of yeast cytochrome b protrudes from the matrix side of the inner membrane consistent with the eight-helix model for the orientation of cytochrome b in the membrane.

Amino Acid Sequence↗
PubMed1999

Chemical orientation to food by the crayfish Orconectes rusticus: influence of hydrodynamics.

Many different organisms orient to chemicals in a variety of habitats. Each of these habitats has a unique hydrodynamic environment that is dependent upon the structure of that habitat. Differences in the hydrodynamics (i.e. turbulence) of an environment will be reflected in the fine-scale structure of chemical signals. To determine what role dispersion dynamics play in influencing orientation behaviour, we studied crayfish searching for food sources in different artificial streams. Streams differed only in substrate composition (sand or cobbles), and the hydrodynamics associated with different substrates were quantified. A detailed analysis of orientation paths showed that crayfish could orient to food sources in streams with either substrate. The most parsimonious explanation is that animals are using information contained in the spatial and temporal distribution of chemicals in the flow to make directional decisions. Crayfish located the source more quickly, spent more time moving, and walked faster while orienting in streams having a cobble substrate compared with those having a sand substrate. These differences between substrates were not seen in control streams. These results show that the hydrodynamics associated with chemical signal structure can greatly influence the temporal properties of orientation to food sources. For crayfish, differences in the turbulent structure of flow may actually increase orientation efficiency by decreasing search time. On a broader scale, these results show that it is important to quantify orientation behaviour in a number of hydrodynamically different environments. Copyright 1999 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed1997

Assessment of g-dependent Cellular Gravitaxis: Determination of Cell Orientation from Locomotion Track

Movement of cells in the gravity field is principally affected in two ways: velocity and orientation. Experimental observation of gravitaxis in large cell populations can document the velocity and orientation of swimming tracks, but orientations of individual cells are not represented at low magnifications. Cell orientations may depart from track orientations due to superposition of sedimentation on cellular propulsion. Here, we show that determination of the sedimentation rate in addition to cell track parameters allows a reconstitution of cell orientation employing geometric principles. Published and original cellular data indicate that gravitactic orientation of cells swimming in the gravity field is superior to that suggested from the experimental tracks. Similar conclusions apply to cells which walk or glide along substrate surfaces. Calculation of cell orientation coefficients provides a basis for determinations of the acceleration-dependence of gravitaxis and for quantitative tests on physical and/or physiological principles of cellular gravitaxis. Copyright 1997 Academic Press Limited

Journal Article↗
PubMed1980

Early lesions of mouse vibrissal follicles:: their influence on dendrite orientation in the cortical barrelfield.

There is a statistically significant order in the tangential orientation of stellate cell dendrites, both spiny and smooth, in layer IV of the barrelfield of the mouse parietal cortex. Neurones situated in a barrel side have most of their dendrites oriented towards the barrel hollow; those situated in the hollow preferentially orient their dendrites parallel to the long axis of the barrel. A quantitative measure of the orientation of individual dendrites in barrelfields of 60-day old mice was obtained using a semi-automatic computer-microscope and a minicomputer. In the same manner, the dendrite orientation of layer IV stellate cells was determined in barrelfields, whose (cytoarchitectonic) pattern had been experimentally altered through lesions of the middle row of the mystacial vibrissal follicles at birth. The dendrites of these cells are oriented in harmony with the novel parcellation of the cortex. Consequently, for cells in the altered areas of the barrelfields, the dendrite orientation is different from that of cells with identical positions in a normal field (see Fig. 8). We tentatively interpret these findings as an adaptation of dendrite orientation to an altered pattern of thalamic input to layer IV that, in turn, is a consequence of the peripheral manipulation.

Animals↗
PubMed1986

The orientation of the cervical vertebral column in unrestrained awake animals. I. Resting position.

The orientation of the cervical vertebral column was studied by X-ray photography of the region containing the head and the neck in nine unrestrained species of vertebrates (man, monkey, cat, rabbit, guinea pig, rat, chicken, frog, lizard). In addition, the orientation of the horizontal semicircular canals was measured in four species using landmarks on the skull. In all vertebrates studied, with the exception of frog and lizard, the general orientation of the cervical vertebral column was vertical when animals were at rest, and not horizontal or oblique as suggested by the macroscopic appearance of the neck. The posture of the animal, whether lying, sitting or standing, had little effect on this general vertical orientation, although some variability was noticed depending on the species. This finding prompted the definition of a resting zone, where the cervical column can take any orientation within a narrow range around a mean position. The cervical vertebral column composes part of the S-shaped structure of the entire vertebral column, with one inflection around the cervico-thoracic (C7/Th1) junction. This feature is already noticable in the lizard. The vertical orientation of the cervical vertebral column is interpreted to provide a stable and energy saving balance of the head. Furthermore, when the head is lowered or raised, the atlanto-occipital and cervico-thoracic junctions are predominantly involved, while the entire cervical column largely preserves its intrinsic configuration. The curved configuration of the cervico-thoracic vertebral column embedded in long spring-like muscles is interpreted to function as a shock absorber. At rest, animals did not hold their heads with the horizontal canals oriented earth horizontally all the time, but often maintained them pitched up by ca. 5 deg, as has been reported for man. At other times, presumably when the vigilance level increased, the horizontal canals were brought into the earth horizontal plane. The vertical orientation of the cervical column results in a vertical positioning of the odontoid process of the axis (second cervical vertebra, C2), which thus provides the axis of rotation for yaw movements of the head. This axis corresponds to that of the horizontal semicircular canals. The vertical organization of the cervical vertebral column in birds and mammals, whether the animal is quadrupedal or bipedal, points to a common organizational principle for eye and head movement systems.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Cortical templates for the self-organization of orientation-specific d- and l-hypercolumns in monkeys and cats.

Blasdel and Salama's sensory maps of orientation-selective edge detectors in the monkey striate cortex can be reduced to an idealized scheme in which orientation hypercolumns of the d- and l-type occur in alternating sequence (Fig. 1). This scheme resolves the apparent contradiction between linear and circular arrangements of successive edge directions in earlier accounts. The actual configuration of hypercolumns is in register with two possible templates for the self-organization of orientation selectivity: the isometric cytochrome oxidase blobs of the colour system, and the anisometric slabs of the ocular dominance system. The centers of the hypercolumns coincide with the blobs. Simulation of cortical self-organization shows this co-incidence even in the absence of template-specific interactions. However, blobs and slabs are symmetrical to these centers, and therefore no templates for the asymmetrical distribution of preferred orientation in the hypercolumns. The present simulation derives the pre-natal formation of an initial scheme from a hypothetical gradient of nervous activity. Post-natal formation, or maturation, of this scheme is achieved by visual experience. Simulation of corresponding interactions between simultaneously activated neurons illustrates both the gain in orientation selectivity (Figs. 2 and 3), and the optimization of farfield diversity and nearfield conformity (Figs. 4 and 5). The results are compatible with the actual distribution of blob-centered d- and l-hypercolumns, iso-orientation modules and orientation fractures in the monkey. A surprisingly similar distribution of blobless d- and l-hypercolumns is expected in the absence of the colour system. Applied to the apparently blobless cortex of the cat, the scheme explains the modulation of deoxyglucose uptake along the iso-orientation bands in a report of Löwel, Freeman, and Singer.

Animals↗
PubMed1991

Distribution of actin filament lengths and their orientation measured by gel electrophoresis in capillaries.

F-actin was electrophoresed in capillary tubes filled with agarose gel. The use of capillary imparted high resistance on the gel allowing the use of high enough concentration of salts to keep F-actin polymerized, and allowed the application of high electric fields without liberating considerable amount of heat. The intensity profile of the electrophoretic band of F-actin showed a peak, which in 1% agarose in the electric field of 17.8 V cm-1 at 0 degree C, migrated at 3.4 cm hr-1. Microscopic observation of actin filaments extracted from different positions along the gel showed that during electrophoresis filaments distributed themselves in such a manner that the longest polymers migrated slowest and the shortest migrated fastest. Using this observation we calculated the weight and number distributions of filament lengths from corresponding experimental intensity profiles. Phalloidin-labelled F-actin oriented in the gel upon application of an electric field. F-actin showed unusual orientational response: it oriented rapidly when the field was applied, but relaxed very slowly when the field was removed. Orientation of F-actin varied within an electrophoretic band, longest polymers showing the best orientation and short oligomers and monomers not orienting at all. The degree of orientation increased with the size of the electric field. When F-actin was labelled with phalloidin before electrophoresis, it was no longer able to migrate in the gel, but the electric field oriented it in the same way as when it was labelled after the electrophoresis. These results show that the electrophoresis of F-actin in agarose fractionates it according to its length, that by using electrophoresis it is possible to rapidly obtain distribution of filament lengths, and that F-actin migrates in agarose by the process of reptation.

Actins↗
PubMed1997

Prehension of objects oriented in three-dimensional space.

When reaching for an object, the proximity of the object, its orientation, and shape should all be correctly estimated well before the hand arrives in contact with it. We were interested in the effects of the object's orientation on manual prehension. Subjects were asked to reach for an object at one of several possible orientations. We found that the trajectory of the hand and its rotation and opening were significantly affected by the object's orientation within the first half of the movement. We also detected a slight delay of the wrist relative to the forearm and a small bias of the orientation of the fingers' tips toward the orientation of the table on which the object lay. Finally, the aperture of the hand was proportional to the physical size of the object, which shows that size constancy was achieved from the variation of the object's orientation. Taken together, these results indicate that the three components of the movement - the transport, rotation, and opening of the hand - have access to a common visual representation of the object's orientation.

Adult↗
PubMed1980

Orientation of pigments and pigment-protein complexes in the green photosynthetic bacterium Prosthecochloris aestuarii.

The orientation of pigments and pigment-protein complexes of the green photosynthetic bacterium Prosthecochloris aestuarii was studied by measurement of linear dichroism spectra at 295 and 100 K. Orientation of intact cells and membrane vesicles (Complex I) was obtained by drying on a glass plate. The photochemically active pigment-protein complexes (photosystem-protein complex and reaction center pigment-protein complex) and the antenna bacteriochlorophyll a protein were oriented by pressing a polyacrylamide gel. The data indicate that the near-infrared transitions (Qy) of bacteriochlorophyll c and most bacteriochlorophyll a molecules have a relatively parallel orientation to the membrane, whereas the Qy transitions of the bacteriochlorophyll a in the antenna protein are oriented predominantly perpendicularly to the membrane. Carotenoids and the Qx transitions (590-620 nm) of bacteriochlorophyll a, not belonging to the bacteriochlorophyll a protein, have a relatively perpendicular orientation to the membrane. The absorption and linear dichroism spectra indicate the existence of different pools of bacteriochlorophyll c in the chlorosomes and of carotenoid and bacteriopheophytin c in the cell membrane. The results suggest that the photosystem-protein and reaction center pigment-protein complexes are oriented with their short axes approximately perpendicular to the plane of the membrane. The symmetry axis of the bacteriochlorophyll a protein has an approximately perpendicular orientation.

Bacterial Proteins↗
PubMed1980

The orientation of the magnetic axes of the membrane-bound iron-sulfur clusters of spinach chloroplasts.

Spinach chloroplast membranes were oriented onto mylar sheets by partial dehydration, and the orientation of the magnetic axes of membrane-bound paramagnetic clusters determined by electron paramagnetic resonance (EPR) spectroscopy. Our results indicate that the reduced Rieske iron-sulfur cluster signal is of orthorhombic symmetry oriented with th gy = 1.90 axis orthogonal to the membrane plane and with the gz = 2.03 axis in the membrane plane; the gx-axis is undetectable, presumably due to its broadness. If the Rieske center is a two-iron iron-sulfur cluster, we conclude that the iron-iron axis lies in the plane of the membrane. Illumination reduces the two bound chloroplast iron-sulfur proteins known as Clusters A and B. Center A is oriented such that gx = 1.86 and gy = 1.94 lie at an angle of about 40, and gz = 2.05 is at approximately 25, to the membrane plane. There are two possible orientations of Cluster B depending on the set of g-values assigned to this cluster. For one set of g-values, gz = 2.04 and gx = 1.89 are oriented in the plane of the membrane while gy = 1.92 is orthogonal to the plane. Alternatively, gz = 2.07 and gy = 1.94 are oriented approximately 50 and 40 to the membrane plane respectively, and gx = 1.80 is in the plane of the membrane. An additional light-induced signal at g = 2.15 oriented orthogonal to the plane is currently unexplained, as are other membrane perpendicular signals seen at g = 2.3 and g = 1.73 in dark-adapted samples.

Cell Membrane↗
PubMed1988

Filamentous phage morphogenetic signal sequence and orientation of DNA in the virion and gene-V protein complex.

The circular single-stranded viral DNA (ssDNA) of filamentous phage is oriented in the virus with a hairpin forming sequence called mos at the leading end of the virion-the end that emerges from the cell first. In the experiments that originally defined mos it was also found to enhance virion production 100-fold, though in other circumstances it has no such effect. Using a new electron-microscopic method, we have ascertained the orientation of ssDNA in phage having mutations involving mos and other viral functions, and also in the intracellular precursor to the virion-a rod-shaped complex between the ssDNA and the phage-encoded gene-V protein, pV. The results show (1) that the ssDNA is oriented in the complex as in the virion, with mos at one end; (2) that orientation is maintained even if assembly is not mediated by the complex; (3) that orientation is manifested in polyphage--abnormal particles in which many unit-length ssDNA molecules are sheathed in a single, extremely long capsid; (4) that orientation is imposed by mos itself (or something very nearby) since it disappears in a mos deletion; and (5) that a 229-base segment including the minus strand of mos is also effective at imposing orientation. On the basis of our findings, we speculate that mos determines orientation in two stages, one imposing an axis on the ssDNA loop during formation of the pV/ssDNA complex, the other imposing a direction on the loop during initiation of particle assembly. The sequence requirements for the two stages may be different.

Bacteriophages↗
PubMed1985

Orientation selectivity in cats and humans assessed by masking.

A two-alternative, spatial forced-choice procedure was used to measure contrast thresholds for detection of sinusoidal gratings that appeared within one-dimensional random noise. The orientation of the noise was symmetrically varied relative to the orientation of the test grating to derive estimates of orientation tuning at two spatial frequencies for both cats and humans. For cats, orientation tuning (half-width at half-strength) averaged 23 deg while for humans tuning averaged 28 deg. Both species displayed narrower tuning at the higher spatial frequency. In addition, evidence is presented that estimates of orientation tuning may be narrower when only one orientation of noise is presented, due to "off-channel" detection strategies. These estimates of orientation tuning are discussed in terms of the orientation selectivity of cortical neurons.

Animals↗
PubMed1991

Orientation discrimination depends on spatial frequency.

Thresholds were measured for discriminating the orientation of sinusoidal gratings of varying spatial frequency, and found to decrease monotonically with increasing spatial frequency. For discrimination of high-contrast (10 times threshold) near-vertical gratings, thresholds ranged from about 1 deg at 0.04 c/deg to 0.5 deg at 0.2 c/deg, after which there was little improvement. At lower contrasts and for discriminations around a mean of 45 deg, thresholds varied more so, and continued to improve until 1 c/deg. The variation of orientation discrimination thresholds with spatial frequency follows a similar trend to the variation in orientation bandwidth of visual units over the same range of spatial frequencies. Thus the present results are consistent with recent "opponent-process" models of orientation discrimination, that predict that thresholds to be limited (at least in part) by the maximum slope of orientation selectivity of visual detectors. That thresholds for high contrast vertical gratings did not improve for frequencies higher than 0.2 c/deg implies that orientation bandwidth and noisiness of oriented detectors may not be the sole factor limiting orientation discrimination, and suggests the existence of more central noise sources.

Contrast Sensitivity↗
PubMed1995

Sensitivity to orientation modulation in micropattern-based textures.

We have measured the sensitivity of the human visual system to sinusoidal modulations of orientation in micropattern-based textured stimuli. The result is the orientation modulation function, or OMF, which describes this sensitivity as a function of the spatial frequency of orientation modulation. We found that the OMF was bandpass with peak sensitivity at spatial frequencies ranging between 0.06 and 0.2 c/deg, depending on the size of the micropatterns. The OMF was found to be scale invariant, that is its position on the spatial frequency axis did not change with viewing distance when spatial frequency was measured in object rather than retinal units. This scale invariance was shown to result from the visual system taking into account the scale rather than the density of the micropatterns as viewing distance was changed. It has been argued by Bergen [(1991) Vision and visual dysfunction (Vol. 10B) New York: Macmillan] that scale invariance in textures is a consequence of the coupling of mechanisms which detect textural features with those which detect local luminance contrasts. We reasoned that Gabor micropattern textures might therefore show narrower OMFs compared to line micropattern textures. However we found no difference in OMF bandwidth between the Gabor and line micropattern textures, suggesting that the line micropatterns were acting as selectively as the Gabor micropatterns for the spatial scale of the mechanisms which detected the orientation modulation. Evidence is presented which suggests that the mechanisms which detected the orientation modulation in our stimuli are non-linear. Finally we showed similar OMFs for sine-wave and square-wave modulations of micropattern orientation, and similar OMFs for modulations of micropattern with orientation about the horizontal and about the vertical, the direction of modulation in both cases being horizontal. The implications of these findings for the mechanisms involved in orientation-defined texture processing is discussed.

Contrast Sensitivity↗
PubMed1996

Color, orientation and cytochrome oxidase reactivity in areas V1, V2 and V4 of macaque monkey visual cortex.

Color and orientation processing in the macaque monkey first segregates into cytochrome oxidase (CO)-rich blobs and -poor interblobs of area V1, from where the two streams flow through areas V2 and V4. This parallel representation is believed to enhance processing speed by compartmentalizing tasks of similar kinds, though our knowledge of the mechanisms is still elementary. We have examined the interaction and separation of color and orientation processing in neurons (n = 569) of the macaque visual cortex (V1, V2, V4) on the basis of microelectrode recordings. In all three areas, neurons selective for midspectral (MS) colors (e.g., yellow, green) were also found to be more orientation selective than those preferring endspectral (ES) colors (e.g., blue, red). The majority of achromatic (AC) cells responsive to bright stimuli were also orientation selective. When locations of cells and penetration columns were correlated with cytochrome oxidase (CO) landmarks in V1 and V2, V1 interblob and V2 interstripe cells were found to be predominantly midspectral and oriented, while V1 blob and V2 thin stripe cells were found to be predominantly endspectral and non-oriented. Cells preferring dark colors were found to cluster in thick stripes in V2, and in columns in V4. Separate clustering of midspectral (MS) and endspectral (ES) systems in V4 was also noted. With the results shown in a companion paper (Behav. Brain Res., 76 (1996) 51-70), the present data indicate that the visual system appears to optimize color and spatial acuity by separating chromatic information into non-oriented endspectral and oriented midspectral components.

Action Potentials↗
PubMedDate not supplied

Flagellar apparatus absolute orientations and the phylogeny of the green algae.

The absolute orientation of the flagellar apparatus in green algal motile cells is a feature of considerable value in studies of green algal systematics and phylogeny. The absolute orientation patterns found in those algae for which this feature is known or can be deduced are reviewed. Counterclockwise absolute orientation occurs in all classes except the Chlorophyceae and is considered primitive, while the clockwise absolute orientation present in most members of the Chlorophyceae is the result of progressive clockwise rotation of components during evolution. Extant intermediates documenting this rotation include Hafniomonas vegetative cells, which show counterclockwise absolute orientation, and Chaetopeltis quadriflagellate zoospores, in which the flagellar apparatus is strictly cruciate except for a slight clockwise offset of the microtubular rootlets. The V-shaped arrangement of the basal bodies in the flagellar apparatus, as well as the presence of proximal sheaths and of two layers of scales on the cell body, further identifies the Chaetopeltis zoospore as a primitive cell type within the Chlorophyceae . Trends towards the exsertion of basal bodies from a flagellar pit, either apically or laterally, the elimination of quadriflagellate cells, and, in the Chlorophyceae , an increasing amount of basal body offset, indicate advancement within the classes. Absolute orientation is conserved during flagellar apparatus replication and development. Events after flagellar apparatus division in the algae studied may be subdivided into component assembly, which is universal and preserves phylogenetically-useful features, and component reorientation, which occurs in relatively few green algae and adapts the flagellar apparatus to specialized functions. From these flagellar apparatus orientation studies, a major reevaluation of evolution within the Chlorophyceae is proposed, with weakly- thalloid algae possessing desmoschisis (e.g. Chaetopeltis ) considered primitive, and most other types, including the Volvocales , considered more advanced. The evolution of wall formation does not preclude the formation of scales in primitive chlorophycean genera. In addition, one or more previously undescribed major lineages may exist within the green algae, including one, the Pleurastrum lineage, whose members possess dorsiventrally -flattened motile cells, counterclockwise absolute orientation of the flagellar apparatus, and a phycoplast at cytokinesis. The Chlorophyceae , the Ulvophyceae , and the Pleurastrum lineage are considered to have a common ancestor that resembled the modern genus Pyramimonas , while the Charophyceae is thought to be of more ancient derivation. Th

Chlorophyta↗
PubMed2004

Acquiring laparoscopic skill proficiency: does orientation matter?

OBJECTIVE: This study was undertaken to determine whether side-on laparoscopic operating orientation alters time-to-skill mastery compared with head-on orientation. STUDY DESIGN: One hundred thirty-one medical students were randomly assigned by operating axis (camera position to operating field) and completed 10 attempts at each of 5 previously validated laparoscopic skills stations. Time-to-completion was recorded for each attempt, generating an orientation and skill station learning curve. Statistical analysis was performed by using repeated measures analysis of variance and linear, polynomial, and logarithmic models with 95% CIs. RESULTS: Sixty-eight students were randomly assigned to head-on orientation and 63 to side-on orientation. Comparing median time-to-completion by station, head-on attempts were faster than side-on attempts for every station. Comparing learning curves by orientation, side-on learning curves were steeper than head-on learning curves for every station, except one. Asymptotes were reached in both strata by the tenth attempt. CONCLUSION: Greater initial disorientation is seen with side-on orientation compared with head-on orientation. This can be overcome with practice.

Clinical Clerkship↗
PubMed2004

Fibroblast responses to cyclic mechanical stretching depend on cell orientation to the stretching direction.

Fibroblasts in intact tendons align with stretching direction, but they tend to orient randomly in healing tendons. Therefore, a question arises: Do fibroblast responses to mechanical stretching depend on their orientation? To address this question, human patellar tendon fibroblasts were grown in custom-made silicone dishes that possess microgrooved culture surfaces. The direction of the microgrooves was either parallel or normal to the direction of cyclic uniaxial stretching. Fibroblasts grown in these microgrooves had a polar morphology and oriented along the direction of the microgrooves regardless of the stretching conditions. Tendon fibroblasts expressed higher levels of alpha-smooth muscle actin when they were oriented parallel to the stretching direction than when they were oriented normal to the stretching direction. Also, cyclic stretching of the fibroblasts perpendicular to their orientation induced a higher activity level of secretory phospholipase A(2) compared with stretching of the cells parallel to their orientation. Thus, these results show that fibroblast responses to mechanical stretching depend on cell orientation to the stretching direction.

Actins↗