Effects of extracellular human immunodeficiency virus type 1 vpr protein in primary rat cortical cell cultures.

Recent evidence suggests that HIV-1 Vpr exists in soluble form in the serum and cerebrospinal fluid (CSF). Further, its abundance in the bloodstream, and the CSF, and its activity on other cell types suggest that it could have an effect on brain activity. Using mixed embryonic rat brain cultures as a model to examine the effects of physiological concentrations of extracellular Vpr protein, Vpr-induced cell death was observed. We also observed similar Vpr-induced effects in enriched primary cortical rat astrocytes, as well as in the C6 glioma cell line. Vpr-induced cell death observed in the astrocytic cells appeared to be caused primarily by a necrotic mechanism, although a few apoptotic nuclei were also present. We did not observe Vpr-induced effects on any primary cortical neurons, although we did observe Vpr-induced cell death in hippocampal neurons and astrocytes. Finally, we observed no cell cycle effects due to extracellular Vpr protein. This data points out that different cell types are affected by the toxic effects of extracellular Vpr protein, and that differential toxic effects of extracellular Vpr protein are observed in similar cell types.

A vitamin E-deficient diet affects nerve regeneration in rats.

Degenerative changes in the neuromuscular system have been found in animals and humans with vitamin E (E) deficiency. This morphologic study examined the effect of dietary E on the regeneration of peripheral nerves in male Sprague-Dawley rats. After feeding an E-sufficient diet (dl-alpha-tocopheryl acetate 50 mg/kg diet) for 6 d, 24 rats were randomly and equally assigned to one of three groups: control (CTRL) fed an E-sufficient diet for 43 d without surgery, normal (NE) fed an E-sufficient diet, or low (LE) fed an E-deficient diet (dl-alpha-tocopheryl acetate 0 mg/kg diet). After 22 d of feeding, NE and LE had surgical compression of the right sciatic nerve and continued eating for 15 d. On day 43, the right triceps surae muscles and a segment of the right sciatic nerve were removed, then all rats were euthanized. The nerve and muscles were processed for morphologic analyses. Presurgery and postsurgery LE ate less food (P < 0.048 and P < 0.001, respectively), which resulted in a lower body weight gain (P < 0.0002). LE had irregularly shaped and less myelinated axons than NE (P < 0.0001) and CTRL (P < 0.0001). The LE plantaris muscle had less type II fibers when compared with NE (P < 0.007) and CTRL (P < 0.03). The results suggest that an E-deficient diet affects food intake, impairs nerve regeneration, and decreases type II fibers, whereas an E-sufficient diet contributes to normal axon regeneration.

Experimental perturbation of the development of sexual size dimorphism in the mouse skeleton.

BALB/c mice were raised for 30 days under nine treatment regimens to determine their effects on sexual dimorphism for size of the femur and of the body. Treatment groups were divided into three experiments: 1) burrowing in a high litter cage; eight times normal gravity for a total of one hour per day; a voluntary exercise control. 2) Unilateral femoral and sciatic nerve ablation; the opposite unoperated leg. 3) Swimming exercise; anabolic steroid; swimming and anabolic steroid; normal exercise control. Traits measured were body weight gain and thirteen femoral characters: moment at failure, length, cross-sectional area, anteroposterior and lateromedial diameters of the cross section, wall thickness for all four quadrants of the shaft, moments of inertia of the cross section in the anteroposterior and lateromedial directions, deflection of the shaft before failure, and total mineral content. Sexual dimorphism of the traits studied varied significantly with treatment. The greatest dimorphism was seen in the controls and the burrowing group. The least dimorphism was seen in the eight times gravity, the steroid, and the exercise and anabolic steroid groups. Reduction in the extent of sexual dimorphism was not correlated with a reduction in the loading regimen but rather seemed mediated by systemic changes in physiology brought on by several factors. Change in the amount of sexual dimorphism for size from the control reflects change in growth rates for males and females in response to different treatments. Change in growth rates of juvenile males and females will likely result in changes in the size of femoral characters and body weight in the adult. Change in the size of femoral characters affects the strength of the femur. In natural populations, a change in the body size of adult males and females could adversely affect reproductive success in groups that normally exhibit strong dimorphism, and hence, select against any conditions that reduce sexual dimorphism. Ordinarily, a significant change in sexual dimorphism--of the magnitude seen here--through natural selection, would take several generations.

Adaptive modeling in a mammalian skeletal model system.

Juvenile BALB/c mice were used as a model system to test the effects of various loading and exercise regimens on the growth and development of femora. Six treatments and three controls were used to document changes in geometric, mechanical, and material properties of the femora associated with strength. In each age-matched experiment, body weight and the strength, length, anterior and posterior diameters, cross-sectional area, moments of inertia in the anteroposterior and lateromedial directions, cortical wall thickness, and mineral content of the femora were assessed and found to vary significantly among treatment groups. An adaptive interpretation of these data was provided by calculating Pearson correlation coefficients between moment at failure (one measure of strength) and each geometric, mechanical and material property of the femora that contributes to strength. We make the assumption that at the termination of the experiment the greater the coordination between changes in strength and changes in the parameters that contribute to strength (the greater the number of correlations), the more adaptively modeled the femora are. Adaptive modeling here refers to the manner in which the femora grow and develop (adapt) under a given treatment regimen. Absolute strength of whole femora was reflected by our measure of adaptive modeling in all groups with one exception. In each experiment, the voluntary exercise controls were the most adaptively modeled. The least adaptively modeled groups also showed a general retardation of growth. It appears that juvenile mouse femora demonstrate a wide range of responses to different conditions of loading and exercise and that some of these changes are likely permanent. Moreover, at least two major variables--1) mechanical loading and 2) glucocorticoid mediated psychological stress--appear to contribute to the differences seen between the treatment groups.

Electromyographic cross-talk within a compartmentalized muscle of the cat.

1. Experiments were conducted to test the extent to which the electromyographic (EMG) activity generated by the activation of single motor units is conducted from one neuromuscular compartment of the cat lateral gastrocnemius (LG) muscle into adjacent compartments. 2. Potentials produced by stimulation of forty-five single motor units were monitored from bipolar fine-wire EMG electrodes which had been implanted either into the centres of each of the four neuromuscular compartments of LG or into regions of the muscle known to lie on the border of contiguous compartments. 3. In all cases single unit potentials could be recorded from the electrodes in the centre of the compartments which clearly identified the compartment of residence of the muscle unit. Regardless of unit type, the amplitude of the potential recorded from electrodes in one compartment was always greater than that recorded from any other compartment. 4. Smaller potentials could be recorded from electrodes in the centre of compartments adjacent to the compartment of residence of the muscle unit. For those motor units where the amplitude of the EMG potentials recorded from the compartment of residence was large, the amplitude of such 'cross-talk' could be greater than the amplitude of potentials recorded from the compartment of residence of smaller motor units. 5. In the case of electrodes placed at compartment boundaries, no clear compartment selectivity of recording of motor unit potentials was evident. 6. These results indicate that great care must be taken in choosing sites of EMG electrode placement when performing kinesiological studies, especially when the amplitude of the EMG activity recorded is of consideration.

Cat triceps surae motor nuclei are organized topologically.

A "compartment nucleus" consists of a group of spinal motoneurons that supply the individual primary branches of a muscle nerve. Each primary muscle nerve branch innervates a specific subvolume or compartment of a muscle. The spatial and size distributions of motoneurons supplying primary nerve branches of the cat medial gastrocnemius muscle were examined to: determine whether or not the medial gastrocnemius motor nucleus is organized in the same manner as that of the lateral gastrocnemius; determine the type of organization which collectively governs the triceps surae (lateral and medial gastrocnemius, soleus) motor nuclei. Individual primary branches of the medial gastrocnemius nerve were exposed to a horseradish peroxidase solution to retrogradely label their cells of origin. Each compartment nucleus was compared with the distribution of the entire medial gastrocnemius motor nucleus. These data were also compared with data on the spatial and size distribution of motoneurons in the lateral gastrocnemius motor nucleus. Results indicate rostrocaudal overlap in the distribution of motoneurons in each compartment nucleus. However, a rostral to caudal topographic arrangement and preferential size distribution for each compartment nucleus exist for proximal to distal muscle compartments. These and other results indicate that the motor nuclei of the entire triceps surae muscle group are organized by a topologic type map.

An anatomical and functional analysis of cat biceps femoris and semitendinosus muscles.

The anatomy, architecture, and innervation patterns of the hamstring muscles, biceps femoris, and semitendinosus were examined in adult cats using microdissection and glycogen-depletion techniques. The biceps femoris muscle consists of two heads. The anterior head, which attaches mainly to the femur, is divided into two parts by the extramuscular branches of its nerve. The posterior head is innervated by a single nerve. Semitendinosus is composed of two heads, one proximal and one distal to a tendinosus inscription, each of which is separately innervated. The extramuscular branches of the nerves to these hamstring muscles thus partition them into innervation subvolumes termed parts. The available evidence suggests that each of the parts of these muscles so innervated is not equivalent to the collections of single motor units that have been described for ankle extensors as neuromuscular compartments. It is quite likely that each of the parts of the hamstring muscles may contain more than one neuromuscular compartment. Using chronically implanted EMG electrodes, the activation patterns of different parts of the hamstring muscles were analyzed during locomotion. The anterior and middle parts of biceps femoris are active during the early stance phase, probably producing hip extensor torque. The posterior part of biceps femoris and semi-tendinosus act most consistently as flexors, during the early swing phase, but also may function in synergy with hip, knee, and ankle joint extensors near the time of foot placement. Greater variability is found in the activity patterns of posterior biceps femoris and semitendinosus with respect to the kinematics of the step cycle than is observed for anterior and middle biceps femoris. It is suggested that this variation may reflect a larger role of sensory feedback in shaping the timing of activity in posterior biceps femoris and semitendinosus than in "nonarticular" muscles.

Thalamic afferents to cytoarchitectonic subdivisions of area 6 on the anterior sigmoid gyrus of the dog: a retrograde and anterograde tracing study.

The cytoarchitecture and thalamic afferents of cortical area 6 located on the anterior sigmoid gyrus were mapped and analyzed in the dog by means of cytoarchitectonic, horseradish peroxidase (HRP), and autoradiographic methods. Cytoarchitectonically, area 6 consists of medial and lateral subdivisions that correspond, respectively, to areas 6a alpha and 6a beta in the cat. In the dog, area 6a alpha is characterized by a wide layer III, the merging of borders between layers III and V, the presence of small-to-medium-size pyramidal cells in layer V, and a pallisade arrangement of cells in layer VI. Area 6a beta appears more stratified, with a relatively acellular layer present between layers V and VI and the presence of large pyramidal cells in layer V. Neither area 6a alpha nor 6a beta contains a layer IV. Data obtained from injections of HRP into areas 6a alpha or 6a beta revealed that labeled thalamic neurons were distributed in a longitudinal band extending from the rostral part of the ventral anterior nucelus (VA) through the caudal part of the mediodorsal nucleus (MD). Labeled cells were observed in the ventral lateral and ventral medial thalamic nuclei as well as in several of the intralaminar nuclei including the central lateral, central medial, parafascicular, and centrum medianum nuclei. A few labeled cells were also located in the suprageniculate nucleus. The densest thalamic labeling was present in VA and MD following injections into area 6a alpha. Equivalent or even larger injections into area 6a beta resulted in much less thalamic labeling. The band of labeled cells also extended into the hypothalamus, zona incerta, amygdala, claustrum, periaqueductal gray of the midbrain, and the nucleus of Darkschewitsch. Results from autoradiographic experiments showed that area 6 subdivisions receive a loosely organized topographic input from VA. Injections of tritiated amino acids were made into selected regions of VA and into the caudal part of MD, areas in which the largest numbers of HRP-labeled cells were located. Area 6a alpha receives afferents primarily from the rostromedial part of VA and the caudal part of MD while area 6a beta receives its principal input from the caudal and lateral parts of VA with minimal input from MD. Axons originating from VA terminate in both layers I and III of area 6 while those originating from the caudal part of MD terminate only in layer III.(ABSTRACT TRUNCATED AT 400 WORDS)

Compartmentalization of the cat lateral gastrocnemius motor nucleus.

The spatial and size distributions of motoneurons which supply the cat lateral gastrocnemius (LG) neuromuscular compartments were examined to determine their anatomical organization. Individual primary muscle nerve branches (PMNB) to LG were isolated by microdissection, cut, and soaked in horseradish peroxidase (HRP). As a control in each case, the entire contralateral lateral gastrocnemius-soleus (LG-S) nerve was similarly cut and soaked. Retrogradely labeled motoneurons were identified, their positions plotted, and their sizes measured on both sides of the cord. Results show that extensive overlap exists in the spatial distribution of motoneurons innervating different PMNBs. Labeled cells supplying each PMNB also vary considerably in their sizes. However, both a highly significant topography-like organization and a preferential size distribution are found between different groups of motoneurons. Neurons which supply proximal compartments occupy more rostral portions of the LG motor nucleus and are among the largest in the pool. Very few small motoneurons innervate proximal compartments. Neurons supplying more distal compartments are distributed in more caudal parts of the pool and contain both large and small cells in relatively equal numbers. If the large cells are alpha and the small cells gamma motoneurons, then proximal compartments are relatively poor in gamma innervation and presumably muscle spindles, and distal compartments are rich in fusimotor innervation.

Compartmentalization of single muscle units in cat lateral gastrocnemius.

The distribution of muscle fibers in single muscle units in cat lateral gastrocnemius (LG) muscle was analyzed using the method of glycogen depletion. Single, type FF motor units in LG were identified by stimulation of dissected ventral rootlets and then activated to produce depletion of glycogen in their innervated muscle fibers. Serial histological sections were made of the entire triceps surae muscles and then reacted for the demonstration of glycogen. Projected tracings of the sections were used to compare the distribution of glycogen depleted muscle fibers to the heads of the LG muscle. Results of analysis of single muscle units indicate that the muscle fibers innervated by single cat LG type FF motor units are distributed to the same regions of muscle which are innervated by primary branches of the nerve to the LG muscle. The single units are thus compartmentalized. These results support the hypothesis that each of the neuromuscular compartments of LG contains a unique population of motor units. It is suggested that compartmentalization of muscles about primary branches of their muscle nerves may from a major organizational principle in the design of skeletal muscles.

Structure of the principal carotenoid pigment of Cellulomonas biazotea.

The yellow-pigmented Cellulomonas biazotea, ATCC 486, contains a mixture of carotenoids. The principal compound is a decapreno carotenoid (C50H72O2) tentatively characterized as 2,2'-bis(4-hydroxy-3-methyl-2-butenyl)-gamma,gamma-carotene on the basis of electronic absorption, infrared, proton magnetic resonance, and mass spectrometries. The carotenoid is presumed to be identical to sarcinaxanthin from Sarcina lutea pro-synon. Micrococcus luteus and, therefore, is isomeric with decaprenoxanthin from Cellulomonas dehydrogenans.

Occurrence of squalene and sterols in Cellulomonas dehydrogenans (Arnaudi 1942) comb. nov. Hester 1971.

The neutral lipid fraction of the photochromogenic, coryneform bacterium Cellulomonas dehydrogenans (Arnaudi 1942) comb. nov. contains the sterol precursor squalene and at least two sterols, cholesterol and beta-sitosterol. The compounds were characterized by mass spectrometry and combination gas-liquid chromatography--mass spectrometry. De novo sterol biosynthetic ability was shown from incorporation of 14C from D-[U-14C]glucose into squalene and the sterol fraction. The squalene concentration approximated 0.002 to 0.005% of the total dry cell weight, and the sterols approximated 0.03 to 0.05%.