ANALYSIS OF THE GEOTROPIC ORIENTATION OF YOUNG RATS. IX

Adult hybrid rats from the cross of races A x B show a total capacity to vary their geotropic performance which is identical with that of their B parents. The proportionate modifiable variability of geotropic orientation also agrees quantitatively with that for the B parents. These relationships are not disturbed by the action of adrenin, which leads to a distortion of the θ vs. α curve. Young rats of the F ₁ generation show a greater proportion of unmodifiable variation of geotropic orientation. It is pointed out that the present findings support the conclusion that the capacity of rats to exhibit variation of geotropic orientation is limited by their genetically determined composition and that the special condition in the young hybrids may be understood as due to a kind of temporary disharmony of developmental events.     

ANALYSIS OF THE GEOTROPIC ORIENTATION OF YOUNG RATS. VII

The intraperitoneal injection of standard young rats of race A with 2/5 cc. of adrenalin chloride 1:50,000 results in increased speed of geotropically oriented creeping upon an inclined surface. It was expected that the effect of such increased frequency of stepping must be analogous to that due to imposition of added loads carried by the rats during geotropic progression. This is verified. The curve connecting θ with log sin α is distorted, under adrenalin, so as to be comparable to that obtained with an added mass of approximately 2.5 gm. upon the young rat''s saddle; the threshold slope of surface for orientation is accordingly lowered, from α = 20° to α = 12.5°; at the new threshold slope of surface the mean orientation angle θ is the same as in the absence of adrenalin at the corresponding threshold slope of surface. The total variation of performance is significantly increased in the injected rats, and at given slope of surface the variation is slightly increased. The proportionate modifiable variation of response is quite unaffected by the distortion of the θ – α curve, and is the same as in standard young A rats untreated or carrying additional loads. It is pointed out that for the consideration of the problem as to whether a given experimental treatment, or a given natural situation, affects in any way the variation of performance of a living system, it is necessary to obtain indices of variability which involve the expression of variation of performance as a function of measured conditions governing the performance.     

ON THE GEOTROPIC ORIENTATION OF HELIX

The snail Helix nemoralis in negatively geotropic creeping orients upward upon an inclined surface until the angle of the path of progression (θ) is related to the tilt of the surface (α) as (Δ sin θ) (Δ sin α) = – const.; θ is very nearly a rectilinear function of log sin α. The precision of orientation (P.E._θ) declines in proportion to increasing sin α, P.E._θ/^θ in proportion to θ. These facts are comprehensible only in terms of the view that the limitation of orientation is controlled by the sensorial equivalence of impressed tensions in the anterior musculature.     

ON THE EQUILIBRATION OF GEOTROPIC AND PHOTOTROPIC EXCITATIONS IN THE RAT

The intensity of light required to just counterbalance geotropic orientation of young rats, with eyelids unopened, is so related to the angle of inclination (α) of the creeping plane that the ratio log I/log sin α is constant. This relationship, and the statistical variability of I as measured at each value of α, may be deduced from the known phototropic and the geotropic conduct as studied separately, and affords proof that in the compounding of the two kinds of excitation the rat is behaving as a machine.     

GEOTROPISM OF AGRIOLIMAX

On an inclined glass plate the slug Agriolimax orients and creeps upward or downward. The angle of orientation on the plane (θ) is proportional to the logarithm of the component of gravity in the creeping plane. The coefficient of variability of the measured values of (θ) decreases linearly as the logarithm at the gravity component in the creeping plane increases. The cosine of the angle of orientation decreases almost directly in proportion to the sine of the angle of inclination of the creeping plane to the horizontal, as previously found for young rats (Crozier and Pincus). But a more satisfactory formulation for the present case shows that the sine of the angle of orientation (θ) decreases in direct proportion to the increase of the reciprocal of the sine of the angle of inclination of the creeping plane. This formulation is derived from the theory that the geotropic orientation is limited by the threshold difference between the pull of the body mass on the mutually inclined longitudinal muscles at the anterior end of the slug.     

ANALYSIS OF THE GEOTROPIC ORIENTATION OF YOUNG RATS. X

The inheritance of elements of geotropic performance in lines of rats (A and B) has been investigated by examining the orientation of young offspring produced in matings of F_1(AxB) with A: Previous studies had shown that the three recognizable groups of receptor elements concerned in geotropic orientation in each of these lines appeared to be inherited in such a way that B groups were dominant with respect to A groups, although this was to a minor extent complicated by influences affecting the variation of orientation as well as the exact form of the curve relating orientation angle (θ) to slope of surface. In the backcross F₁ x A, therefore, at least eight different types of curves were to be expected. These are in fact identifiable among the forty-one individuals carefully studied. Their classification is concordant with the behavior of the respective indices of variation of θ, for which an interpretation has been provided. The basic result is, therefore, that the three receptor groups of excitation units are inherited independently, and alternatively as regards the members of a homologous pair, and that rather simple dominance relations obtain between homologous groups from the two races, namely that a B effect is dominant over the homologous A effect. This interpretation has been tested in various ways, and is in principle completely consistent with the results of a similar experiment involving races A and K.     

ANALYSIS OF THE GEOTROPIC ORIENTATION OF YOUNG RATS. II

1. Equations describing the geotropic orientation of young rats as a function of the inclination of the surface on which creeping take place, under standardized conditions, are found to be of similar form but with different values of the contained constants, when several different, genetically stabilized lines or races are compared. The values of these constants are characteristic for the several races. 2. The biological "reality" of the differences between young rats of two races, as given mathematical form in terms of these parameters and coefficients, can be submitted to radical test by investigating their behavior in inheritance. A simple result favorable to the inquiry would be decisive; a complex, non-clear result would not however be definitely unfavorable to the view that "real" differences in behavior are in question. The actual result is of a kind demonstrating (a) the efficiency of the original formulations, and (b), at the same time, the definite inheritance of certain quantitative aspects of geotropic behavior. 3. On the assumption that orientation on a sloping surface is achieved when, within a threshold difference, the tension-excitations on the two sides of the body (legs) are the same, the angle of oriented progression (θ) can be taken as a direct measure of the total excitation. This is consistent with the equation, accurately obeyed by our initial races, Δ cos θ/Δ sin α = - const., where α is the slope of the surface. 4. The total excitation of tension-receptors must be regarded as involving, over a gross interval of time, (1) the total array of receptors with thresholds below a certain value, a function of the stretching force, and (2) the frequency of change of tension. The latter, largely determined (it is assumed) by the frequency of stepping, should be proportional to the speed of progression. This speed is directly proportional to log sin α. Hence Δθ/Δ log sin α. plotted against sin α, should give a picture of the distribution of effective thresholds among the available tension-receptors in terms of the exciting component of gravity. For the races investigated this distribution can be resolved in each case into three groups. 5. A "variability number" is employed which permits the demonstration that the variability of θ as measured is (1) definitely controlled by α, and is (2) a characteristic number for each of the pure races used. 6. By attaching a weight to rats of one race it is found that Δθ/Δα is modified in a manner concordant with the assumption that the three "groups of sense organs" are in fact discrete. 7. In race K these three groups (I, II, III) are large, in race A, small (i, ii, iii). F₁ rats of the cross between these two races show i, ii, III. 8. F₁ individuals back-crossed to A give in the progeny two sorts of individuals, in equal numbers: i, ii, III and i, ii, iii. 9. F₁ individuals back-crossed to K are expected to give in the progeny four types of individuals, I, II; i, II; I, ii; i, ii. In the numbers available these classes are reasonably clear, and occur with equal frequency. 10. It is pointed out that these considerations imply a mode of definition of a gene somewhat different from that commonly employed by tacit assumption; namely, a definition of the effect in inheritance as a function of some controlling, independent variable.     

THE GEOTROPIC RESPONSE IN ASTERINA

Upon a surface inclined at angle α Asterina gibbosa orients upward during negatively geotropic creeping until the average angle (θ) of the path is such that Δ sin θ/Δ sin α = const. This is true also in positively geotropic movement. The direction of orientation may be temporarily reversed by mechanical disturbance. The variation of θ is greater at low slopes. Tests with directed impressed pulls, due to an attached cork float, show that the pull upon the tube feet is of primary consequence for the determination of θ. When the component of gravitational pull in the direction of movement reaches a fraction of the total pull which is proportional to the gravitational vector parallel to the surface, the laterally acting component is ineffective. On this basis, it follows that Δ sin θ/Δ sin α = const.     

GEOTROPIC ORIENTATION IN ARTHROPODS : I. MALACOSOMA LARV?.

The geotropic orientation of caterpillars of Malacosoma americana during progression upon a surface inclined at angle α to the horizontal is such that the path makes an average angle θ upward on the plane, of a magnitude proportional to log sin α. More precisely, the product (sin α) (sin θ) is constant. This is traced to the fluctuation of the pull of the head region upon the lateral musculature of the upper side during the side to side swinging implicated in progression.     

THE PHOTOTROPIC EXCITATION OF LIMAX

The photic orientation of Limax creeping geotropically upon a vertical plate is such that the phototropic vector determining the angular deflection β from the vertical path is proportional to log I. This is proved by the fact that with horizontal illumination tan β is directly proportional to log I; with non-horizontal light rays from a small source the ratio See PDF for Equation is directly proportional to log I (where A = the angle between light rays and the path of orientation), the vector diagram of the field of excitation being in this case not a right-angled triangle.     

THE USE OF SHADOW-CASTING TECHNIQUE FOR MEASUREMENT OF THE WIDTH OF ELONGATED PARTICLES

A method is described for the estimation of the true width of fibrillar or rod-like structures from electron micrographs of metal-shadowed preparations. The method is based on variations in the image width as a function of the angle (β) between the long axis of the fibril and the direction of the shadow in the plane of the preparation. The image width when β = 0° practically represents the real width of the elongated particle but is often indistinguishable from the background. The fibril image width is conveniently measured at β values between 15° and 90°. The true width is obtained by plotting the image width versus sin β and extrapolating to β = 0°. Latex spheres are sprayed with the fibrils or rods to indicate the direction of shadow. Tobacco mosaic virus (TMV) was used as a model structure because of its known constant diameter of 150 A (5). The width (in the case of TMV equal to the diameter) found by the present method was 150 A ± 8 A.     

GEOTROPIC ORIENTATION IN ARTHROPODS : III. THE FIDDLER CRAB UCA.

On an inclined surface the fiddler crab Uca pugnax, during sidewise progression, orients upward through an angle θ on the surface. The extent of negatively geotropic orientation (θ) is a rectilinear function of sin α, where α is the inclination of the surface to the horizontal. This equation differs from that describing the geotropic orientation of various other animals. The difference is traced to the fact that from an initial position with the transverse axis of the body horizontal the crab is required to turn upward to an extent such that the vertical line from its center of gravity pierces the inclined surface within the base of support provided by the legs. This leads to the equation sec θ/tan α = const., which is obeyed within the limits of precision of the measurements. This type of control of geotropic orientation represents an extension of the "muscle tension theory," and is in no sense in conflict with this view. The assumptions underlying the analytical expression connecting θ and α are verified by the asymmetry in the orientation of male fiddlers, which is shown to be due to the presence of the enlarged chela and which disappears when the claws are removed.     

Angle of Inclination of Tank-Treading Red Cells: Dependence on Shear Rate and Suspending Medium

Red cells suspended in solutions much more viscous than blood plasma assume an almost steady-state orientation when sheared above a threshold value of shear rate. This orientation is a consequence of the motion of the membrane around the red cell called tank-treading. Observed along the undisturbed vorticity of the shear flow, tank-treading red cells appear as slender bodies. Their orientation can be quantified as an angle of inclination (θ) of the major axis with respect to the undisturbed flow direction. We measured θ using solution viscosities (η₀) and shear rates (γ˙) covering one and three orders of magnitude, respectively. At the lower values of η₀, θ was almost independent of γ˙. At the higher values of η₀, θ displayed a maximum at intermediate shear rates. The respective maximal values of θ increased by ∼10° from 10.7 to 104 mPas. After accounting for the absent membrane viscosity in models by using an increased cytoplasmic viscosity, their predictions of θ agree qualitatively with our data. Comparison of the observed variation of θ at constant γ˙ with model results suggests a change in the reference configuration of the shear stiffness of the membrane.     

ORIENTATION IN COMPOUND FIELDS OF EXCITATION; PHOTIC ADAPTATION IN PHOTOTROPISM

During upward geotropic orientation upon a vertical plate the slug Agriolimax creeps vertically, in darkness. Horizontal light from one side produces orientation of dark-adapted slugs away from the vertical path, through an angle (β). The magnitude of this angle is a function of the light intensity and of time. The moderately rapid course of light adaptation is followed by measurements of β at fixed intervals. Simple assumptions as to the nature of the orienting forces lead to the conclusion that the logarithm of the tangent of β should decrease linearly with time, and that the rate of the decrease should vary directly with the logarithm of the light intensity. Both expectations are adequately realized. Certain implications of these results for behavior analysis are pointed out.     

THE ORIENTATION OF ANIMALS BY OPPOSED BEAMS OF LIGHT

When orientation is attained under the influence of beams of parallel light opposed at 180° the deflection θ from a path at right angles to the beams is given by tan See PDF for Equation, where I ₁ and I ₂ are the photic intensities and H is the average angle between the photoreceptive surfaces. This expression is independent of the units in which I is measured, and holds whether the primary photosensory effect is proportional to I or to log I. When photokinetic side-to-side motions of the head occur, H decreases with increasing total acting light intensity, but increases if higher total light intensity restricts the amplitude of random movements; in each case, H is very nearly proportional to log I ₁ I ₂. For beams of light at 90°, See PDF for Equation. The application of these equations to some particular instances is discussed, and it is shown why certain simpler empirical formulæ previously found by others yield fair concordance with the experimental data. The result is thus in complete accord with the tropism theory, since the equations are based simply on the assumption that when orientation is attained photic excitation is the same on the two sides.     

GEOTROPIC ORIENTATION IN ARTHROPODS : II. TETRAOPES.

The creeping of the beetle Tetraopes tetraopthalmus during negatively geotropic orientation shows the angles of orientation (θ) on a surface inclined at α° to the horizontal to be proportional to sin α. The direction of orientation easily suffers temporary reversal to positive as result of handling. Mechanical stability during upward progression should be just possible when K ₁ cot α = K ₂ sin θ + K ₃ cos θ, the weight of the body being supported on the tripod formed by the legs on either side and by the posterior tip of the abdomen. Lack of this stability produces tensions on the legs through (1) the bilaterally distributed pull of the body mass on the legs, and (2) the torque on the legs due to the weight of the abdomen. The downward gravitational displacement of the tip of the abdomen causes K ₂ and K ₃ in the preceding formula to be functions of α. These relations have been tested in detail by shifting the location of the center of gravity, by attaching additional masses anteriorly and posteriorly, and by decreasing the total load through amputation of the abdomen; the latter operation changes the conditions for stability. Different formulæ are thus obtained (cf. earlier papers) for the orientation of animals in which the mechanics of progression and the method of support of the body weight on an inclined surface are not the same. This demonstrates in a direct way that the respective empirical equations cannot be regarded as accidents. The results are in essence the same as that already obtained with young mammals. The diversity of equations required for the physically unlike cases merely strengthens the conception of geotropic orientation as limited by the tensions applied to the musculature of the body (caterpillars, slugs) or of appendages (beetles, and certain other forms) when the body is supported upon an inclined surface, since equations respectively pertaining to the several instances, and satisfactorily describing the observations, are deduced on this basis.     

Propagation Speed in Myelinated Nerve: II. Theoretical Dependence on External Na+ and on Temperature

The Hodgkin-Huxley (H.H.) equations modified by Dodge for Rana pipiens myelinated nerve have been solved to determine how well the theory predicts the effects of changes of temperature and [Na⁺]₀ on propagation. Conduction speed θ was found to have an approximately exponential dependence on temperature as was found experimentally, but the theoretical temperature coefficient (Q₁₀) was low; 1.5 compared with the experimental finding of 2.95. θ was found to be a linear function of log ([Na⁺]₀) in contrast to the experimental finding of a square root dependence on [Na⁺]₀. θ is 50% greater at one-fourth normal [Na⁺]₀ than the theory predicts. The difference between the theoretical θ([Na⁺]₀) and the experimental θ([Na⁺]₀) is probably due to an imprecisely known variation of parameters and not to a fundamental inadequacy of the theory.     

The DNA polymerase III holoenzyme contains γ and is not a trimeric polymerase

There is widespread agreement that the clamp loader of the Escherichia coli replicase has the composition DnaX₃δδ’χψ. Two DnaX proteins exist in E. coli, full length τ and a truncated γ that is created by ribosomal frameshifting. τ binds DNA polymerase III tightly; γ does not. There is a controversy as to whether or not DNA polymerase III holoenzyme (Pol III HE) contains γ. A three-τ form of Pol III HE would contain three Pol IIIs. Proponents of the three-τ hypothesis have claimed that γ found in Pol III HE might be a proteolysis product of τ. To resolve this controversy, we constructed a strain that expressed only τ from a mutated chromosomal dnaX. γ containing a C-terminal biotinylation tag (γ-C_tag) was provided in trans at physiological levels from a plasmid. A 2000-fold purification of Pol III* (all Pol III HE subunits except β) from this strain contained one molecule of γ-C_tag per Pol III* assembly, indicating that the dominant form of Pol III* in cells is Pol III₂τ₂ γδδ’χψ. Revealing a role for γ in cells, mutants that express only τ display sensitivity to ultraviolet light and reduction in DNA Pol IV-dependent mutagenesis associated with double-strand-break repair, and impaired maintenance of an F’ episome.     

Cardiac Dysfunction Induced by Obesity Is Not Related to β-Adrenergic System Impairment at the Receptor-Signalling Pathway

Obesity has been shown to impair myocardial performance. Some factors have been suggested as responsible for possible cardiac abnormalities in models of obesity, among them beta-adrenergic (βA) system, an important mechanism of regulation of myocardial contraction and relaxation. The objective of present study was to evaluate the involvement of βA system components in myocardial dysfunction induced by obesity. Thirty-day-old male Wistar rats were distributed in control (C, n = 25) and obese (Ob, n = 25) groups. The C group was fed a standard diet and Ob group was fed four unsaturated high-fat diets for 15 weeks. Cardiac function was evaluated by isolated papillary muscle preparation and βA system evaluated by using cumulative concentrations of isoproterenol and Western blot. After 15 weeks, the Ob rats developed higher adiposity index than C rats and several comorbidities; however, were not associated with changes in systolic blood pressure. Obesity caused structural changes and the myocardial responsiveness to post-rest contraction stimulus and increased extracellular calcium (Ca²⁺) was compromised. There were no changes in cardiac function between groups after βA stimulation. The obesity was not accompanied by changes in protein expression of G protein subunit alpha (Gsα) and βA receptors (β₁AR and β₂AR). In conclusion, the myocardial dysfunction caused by unsaturated high-fat diet-induced obesity, after 15 weeks, is not related to βAR system impairment at the receptor-signalling pathway.