The structural specificity of factors in the chemical regulation of muscle tonus at the spinal cord level

The chemical factors of the postural asymmetry (FPA) were studied on the recipients without hemispheres. The structural specificity of the fore and hind limb spinal centers regulation in normal and damaged CNS was observed. In the normal CNS this structural specificity was displayed in the selective activation of the cross-situated hemicenters in the cervical and lumbal regions (for example in the left part of the cervical and in the right part of the lumbal region). In the case of the unilateral lesion of the central motor systems the normal pattern of chemical structural specificity was modified by activation of FPA selectively acting on the partly denervated regions of the spinal cord.     

Distribution and subtype determination ofβ-receptors in the spinal cord of the adult rat

1. We determined the number of beta-receptors in the whole spinal cord of the adult rat and in the cervical, thoracal, and lumbal/sacral parts. 2. The undivided spinal cord contains 47 +/- 10 fmol/mg beta-receptors (KD = 2066 +/- 982 pmol/liter), and the cervical part of the spinal cord contains 53 +/- 8 fmol/mg protein (KD = 3224 +/- 1775 pmol/liter). The thoracal part shows 40 +/- 1 fmol/mg protein (KD = 3229 +/- 104 pmol/liter), and the lumbal/sacral spinal cord contains 48 +/- 8 fmol/mg protein (KD = 3610 +/- 1610 pmol/liter). 3. Competitive inhibition studies with l-practolol, dl-atenolol, and ICI 118,551 were performed and we calculated by a computer program in the whole spinal cord the following ratio of beta-receptor subtypes: 80 +/- 5% Beta 1-receptors and 20 +/- 5% beta 2-receptors. 4. The basal and (-)-isoproterenol- and NaF-stimulated activity of adenylate cyclase was highest in the cervical part of the spinal cord and equally distributed between the thoracal and the lumbal/sacral parts. 5. The whole synaptosomal protein of the cervical part of the spinal cord contained 132 +/- 20 fmol, the thoracal part 117 +/- 3 fmol, and the lumbal/sacral part 133 +/- 22 fmol.     

Impairment of the mitochondrial respiratory enzyme activity triggers sequential activation of apoptosis-inducing factor-dependent and caspase-dependent signaling pathways to induce apoptosis after spinal cord injury

The mitochondrion participates in caspase-independent or caspase-dependent apoptotic pathways through the release of apoptosis-inducing factor or cytochrome c. Whether both mitochondrial apoptotic cascades are triggered in the injured spinal cord remains unknown. Here, we demonstrated that neurons, astrocytes and microglia in spinal segments proximal to a complete spinal cord transection underwent two phases of apoptotic cell death. The early phase of high-molecular weight (HMW) DNA fragmentation was associated with nuclear translocation of apoptosis-inducing factor, reduction in mitochondrial respiratory chain enzyme activity and decrease in cellular ATP concentration. The delayed phase of low-molecular weight (LMW) DNA fragmentation was accompanied by cytosolic release of cytochrome c , activation of caspases 9 and 3, and resumption of mitochondrial respiratory functions and ATP contents. Microinfusion of coenzyme Q₁₀, an electron carrier in mitochondrial respiratory chain, into the epicenter of the transected spinal cord attenuated both phases of induced apoptosis, and reversed the elicited mitochondrial dysfunction, bioenergetic failure, and activation of apoptosis-inducing factor, cytochrome c , or caspases 9 and 3. We conclude that mitochondrial dysfunction after spinal cord transection represents the initiating cellular events that trigger the sequential activation of apoptosis-inducing factor-dependent and caspase-dependent signaling cascades, leading to apoptotic cell death in the injured spinal cord.     

Dynamics of the activity of the postural asymmetry factor after unilateral damage to the motor area of the cortex

The authors studied the time-course of functional rearrangements of the segmental apparatus after unilateral injury of the rat motor cortex. It was found that one day after injury the postural asymmetry of the hind limbs was fixed by the lumbal region of the spinal cord. This functional state of the segmental apparatus lasted 10 days after injury in the presence of the maximal activity of postural asymmetry factor (PAF) in the CSF and increasing activity of the factor in the brain tissue. Recovery of the segmental apparatus to symmetrical function by the end of the third week following injury was accompanied by PAF inactivation.     

Characteristics of the development of postural asymmetry in rats as affected by the opioids met-enkephalin and the kappa-agonist bremazocine

It has been found that bremazocine and met-enkephalin induce postural asymmetry in spinal rats under subarachnoidal and intravenous administration. Intravenous administration of bremazocine to intact animals--even if after it (an hour later) their spinal cord is sectioned--produces no asymmetry, i. e. the spinal cord section is necessary for asymmetry development. The magnitude of postural asymmetry and the side of limb flexion are not constant for each animal, but they change in time. Though, on the average, the percent of asymmetric animals and the ratio of left and right flexions in each group of animals are practically constant. When the spinal cord is sectioned at the T1-T4 level, the bremazocine and metenkephalin induce mainly the right-leg flexion: when the section is made at the T5-T9 level, the left-leg is bent, i. e. the flexion side depends on the level of the section. It is suggested that the ability of opioids to induce postural asymmetry is based on lateralization of opioid receptors in the rat spinal cord.     

Met-enkephalin-induced release into the blood of a factor causing postural asymmetry

Met- and Leu-enkephalin applied subarachnoidally into the rostral portion of a transected spinal cord (at the T6-T7 level) induce postural asymmetry of the hind limbs in rats, Met-enkephalin being predominantly responsible for the flexion of the right, and Leu-enkephalin of the left, hind leg. The blood serum of rats injected with Met-enkephalin contains a factor which, when administered subarachnoidally into the caudal portion of the transected spinal cord, is capable of inducing the hind limb postural asymmetry--predominantly, with the right leg flexion. This factor is inactivated by papain and differs from Met- and Leu-enkephalin in chromatographic properties. Apparently, Met-enkephalin induces the release of a peptide factor into the blood, from the brain or organs innervated by the neurons lying above the cut. It is then carried with the blood to the hind limbs and effects the hind limb postural asymmetry.     

The effect of the protective antigen of Bacillus anthracis on the formation of immunity under the action of live anthrax vaccines

The preparation of low-molecular protective antigen (PA) isolated from strain 34F2 (Sterne) and having a molecular weight of 34 and 51 kD, unlike the preparation of high-molecular PA with a molecular weight of 87 kD, suppressed the formation of acquired resistance to anthrax when introduced into guinea pigs in mixture with live spores of strains of STI, 34F2 and new vaccine strain 228/8; this phenomenon was mainly accompanied by a decrease in the level of antibodies to lethal factor (LF) nad in the antitoxic activity of blood serum. The immunosuppressing action of low-molecular PA depended on the kind of vaccine strain introduced together with this antigen, which suggested the existence of differences in the ligand determinants of strains 34F2 and STI. In contrast to high-molecular PA, low-molecular PA blocked the action of the lethal mixture of PA and LF on the culture of peritoneal mononuclear phagocytes of CBA mice. The competitive relationships between low-molecular PA and high-molecular PA are discussed.     

Opioids evoke postural asymmetry in rats: the side of the flexed paw depends on the type of opioid agonist

Opioid kappa-agonists bremazocine and dynorphin (1-13), sigma-agonist SKF 10.047 and delta-agonist D-Ala2, D-Leu5-enkephalin (DADL) induce postural asymmetry of rats hind limbs under subarachnoidal administration below the level of spinal cord section (T3-T4). The side of the flexed leg depends on the opioid agonist type: bremazocine and dynorphin (1-13) induce predominantly right flexion. SKF 10.047--the left flexion, but not in all doses, DADL--in small doses (1 and 100 pg per animal)--of the right one, in larger doses (up to 10 ng per animal)--of the left one. Saline and opiate mu-agonist morphine do not induce postural asymmetry. Opiate antagonist naloxone prevents asymmetry development when injected prior opioid agonists, and also decreases the number of asymmetries induced by these agonists. Naloxone alone does not influence the per cent of animals with pose asymmetry. The opioid receptors are involved in asymmetry development. The revealed ability of opioid kappa-, delta- and sigma-agonists may be based on lateralization of opioid receptors in the rat spinal cord.     

Effect of unilateral motor cortex ablation on activity of choline acetyltransferase and levels of amino acid transmitter candidates in the spinal cord of adult monkeys

Evidence thatl-glutamate is a neurotransmitter of corticofugal fibers was sought by measuring changes in several biochemical markers of neurotransmitter function in discrete regions of spinal cord after ablation of sensorimotor cortex in monkeys. One and five weeks after unilateral cortical ablation, samples from six areas of spinal cord (ventral, lateral and dorsal regions of the left and right sides) were analysed for choline acetyltransferase (ChAT) activity and contents of amino acid transmitter candidates-glutamic acid (Glu), aspartic acid (Asp), glycine (Gly), taurine (Tau) and -aminobutyric acid (GABA). During one to five weeks after unilateral cortical ablation of the monkey, prolonged hemiplegia in the contralateral side was observed. Histological examination of the spinal cord 5 weeks after unilateral (left) cortical ablation showed no apparent change in either control (ipsilateral, left) or affected (contralateral, right) sides of the cord as examined by the Klüver-Barrera method. The ChAT activity as a cholinergic marker was scarcely changed in any region of either left (control) or right (affected) side of the spinal cord at one and five weeks after unilateral (left side) ablation of the motor cortex. Amino acid levels in each region of the spinal cord were not significantly changed one week after unilateral ablation of the motor cortex. However, a significant decrease of Glu content was observed in the lateral column of the affected (right) side compared to the control (left) side of cervical and lumbar cord five weeks after cortical ablation of the left motor area. No concomitant alterations of other amino acids were detected. These data strongly suggest thatl-Glu is a neurotransmitter for corticofugal pyramidal tract fibers to anterior horn secondary neurons related to motor control activity in monkey spinal cord.     

The ontogeny and phylogenetic conservation of MAP 2 forms

Summary In the adult rat brain, MAP 2 is a high-molecular weight protein that is highly concentrated in dendrites. Immunoblots of homogenates of developing rat brain have indicated that a low-molecular weight form of MAP 2, MAP 2 c, is transiently expressed as the brain is undergoing morphogenesis. Using MAP 2-specific monoclonal antibodies, we have demonstrated that the compartmentalization of high-molecular weight MAP 2 and the developmental regulation of MAP 2 are conserved in mammalian, avian, and amphibian brain. We have also determined the distribution of MAP 2 c in developing neuronal tissue. MAP 2 c appears before high-molecular weight MAP 2 in developing neurons, and in contrast to the dendrite-specific high-molecular weight forms of MAP 2, MAP 2 c is present in axons and glia. We have also shown that MAP 2 c is present in the adult rat retina, where it is concentrated in regenerative photosensitive cells. The transient expression of MAP 2 c in the developing brain of three species as well as in adult photosensitive cells suggests a role for this protein in neurite growth and plasticity.Abbreviations MAP microtubule-associated protein - E embryonic day - P postnatal day     

Serum factors stimulating DNA synthesis in the isolated nuclear system from rat liver

³H-TTP incorporation into DNA by the isolated rat liver nuclei was stimulated by the rat serum in proportion to its concentration. Dialysis and gel-filtration of the serum indicated the presence of two factors: one is low-molecular and another is high-molecular. The high-molecular factor is thermolabile while the low-molecular one is thermostable. The latter is resistant to pronase-treatment and can not be adsorbed on charcoal. The sera from normal and partially hepatectomized rats showed similar stimulatory effect.     

Biochemical processes at the stage of withering during black tea production

We determined the molecular weight and some properties of multiple forms of phenol oxidase from tea leaves and four other perennial plants. It was shown that multiple high- and low-molecular forms of phenol oxidase differed in substrate specificity. Low-molecular forms of the enzyme mostly demonstrated hydroxylase activity, while high-molecular forms showed catechol oxidase activity. It was revealed that the withering stage of black tea production is accompanied by the formation of only high-molecular forms of phenol oxidase, which possess catechol oxidase activity crucial for the procurement of oxidative reactions and the quality of the product.     

Spinal regions involved in baroreflex control of renal sympathetic nerve activity in the rat

Spinal cord injury causes debilitating cardiovascular disturbances. The etiology of these disturbances remains obscure, partly because the locations of spinal cord pathways important for sympathetic control of cardiovascular function have not been thoroughly studied. To elucidate these pathways, we examined regions of the thoracic spinal cord important for reflex sympathetic control of arterial pressure (AP). In anesthetized rats, baroreceptor relationships between pharmacologically induced changes in AP and changes in left renal sympathetic nerve activity (RSNA) were generated in spinally intact rats and after acute surgical hemisection of either the dorsal, left, or right T8 spinal cord. None of these individual spinal lesions prevented the baroreceptor-mediated increases in RSNA caused by decreases in AP. Thus, baroreceptor-mediated increases in RSNA in rats are mediated by relatively diffuse, bilateral, descending, excitatory projections. The ability to reduce RSNA at increased AP was impaired after both dorsal and left hemisections, and baroreceptor gain was significantly decreased. Baroreceptor-induced maximum decreases in RSNA were not affected by right hemisections. However, baroreflex gain was impaired. Because both dorsal and left hemisections, but not right hemisections, attenuated the decrease in RSNA at elevated AP, we conclude that pathways involved in the tonic inhibition of spinal sources of sympathetic activity descend ipsilaterally in the dorsal spinal cord. Our results show that many lesions that do not fully transect the spinal cord spare portions of both descending excitatory pathways that may prevent orthostatic hypotension and descending inhibitory pathways that reduce the incidence of autonomic dysreflexia.     

Contents of extracellular DNA in blood of irradiated rats]

Intact rat plasma contains high-molecular DNA which moves as a single fraction in 0.5% agarose electrophoresis. As early as 2-5 hours after gamma-irradiation in a dose of 1-100 Gy there appears low-molecular DNA (about 180 nucleotide pairs), the amount of which directly correlates with the irradiation dose 5 hours after the exposure. Blot-hybridization showed that low-molecular DNA has no common nucleotide sequences with high-molecular DNA, though has sites similar to genome repeat sequences.     

Effect of hypothermia on subcellular distribution and various physico-chemical properties of cathepsin D in rat brain

The total cooling ef rats down to the rectal temperature 30 degrees and 20 degrees C does not change significantly the ratio of the relative specific activity of cathepsin D in subcellular fractions of the rat brain. The gel chromatographic analysis of heterogeneity of cathepsin D molecular forms in subcellular fractions established the presence of a high-molecular (in the fractions of lysosome and microsome mitochondria) and a low-molecular (in the fractions of lysosome and cytosol mitochondria) enzyme forms. Under hypothermia (20 degrees C) in the brain cytosol fraction there arises a minor zone of the cathepsin D activity corresponding to the high-molecular enzyme form.     

Astrocyte death in organotypic cultures of fetal spinal cord from rats with unilateral micromelia

Growth and differentiation of neurons and glia in spinal cord explants of 16 days old rat fetuses with teratogen-induced left-sided micromelia were studied. Progressive destruction of astrocytes that differentiate in interstitial zone of cultures was observed in 37% of explants of the left side, while the development was normal in cultures of the right side. Possible mechanisms leading to destruction of astrocytes in cultures of spinal cord regions that innervate anomalous limbs are discussed.     

Asymmetric changes of the muscle tonus of the hindlimbs of rats subjected to administration of extracts from the left and right brain hemispheres

Suboccipital administration to rats of the solutions of substances extracted from the left and right halves of the brain with 1 M hot acetic acid gives rise to humping, restricts animals' mobility and induces asymmetric changes in the muscle tone of the hind limbs. The hind limb may be extended back or aside at the same side where brain extraction was performed and may retain this position for 25-30 sec. Proteolysis of the extracts or preliminary administration of nalorphine to the animals abolished the occurrence of, while suboccipital administration of naloxon removed the extract-evoked changes in the muscle tone. After application of the extracts to the spinal cord of the hordotomized (Th5-Th7) rats the electrical activity rose at the same side where brain extraction was performed, primarily in the m. quadriceps. It is assumed that active factors extracted from the left and right halves of the brain are different; they are endogenous opioid peptides or else their effects on the activity of spinal motoneurons are mediated via the opiate systems.     

The chemical nature of the fluorescing products accumulating in the lipids of the crystalline lenses of mice with hereditary cataract

The content of diene conjugates (lipid hydroperoxides) was shown to be significantly higher in lipids extracted from the lenses of mice with hereditary cataract than in the controls. The same holds true for characteristics of fluorescence of the end-product of lipid peroxidation. Two (low- and high-molecular weight) peaks were detected in chromatographic lipid profile of cataract lenses measured by fluorescence on Sephadex LH-20 column, whereas only one (high-molecular weight) peak was found in lipids from normal lenses. It was established that high-molecular weight fluorescent fractions corresponded to lipid components of lipofuscin-like pigments. NMR and mass spectrometry of low-molecular weight fractions suggested that they contained predominantly products of free radical oxidation of long chain polyunsaturated fatty acids (C22:6).     

LIGHT-HARVESTING PIGMENT-PROTEIN COMPLEXES OF THE ULVOPHYCEAE (CHLOROPHYTA):CHARACTERIZATION AND PHYLOGENETIC SIGNIFICANCE1

Photosystem II light-harvesting complexes were isolated from a number of ulvophycean algae. Some of these light-harvesting complexes displayed unusual features, most notably a high apparent molecular weight (ca. 58,000) when isolated by lithium doderyl sulfate polyarrylamide gel electrophoresis. Other ulvophycean light-harvesting complexes had a low-molecular weight (ca. 30,000). The distribution of the high-molecular weight complex was limited to certain members of the Caulerpales and Blastophysa rhizopus (Siphanocladales). Within the Caulerpales, there were also spectral differences between the high-molecular weight and low-molecular weight light-harvesting complex types. The differences in light-harvesting complexes in the Ulvophyceae suggest that there are two lines of evolution in the Caulerpales and that Blastophysa may be an intermediate between the Siphon-ocladales and the Caulerpales.     

Asymmetric distribution of peptide regulators of muscle tonus and substance P in the spinal cord of rats with unilateral hyperactivity of the lumbar enlargement neurons

The injection of tetanus toxin in m. gastrocnemius of the left or right hind limb of rats evokes ipsilateral hyperactivity of lumbar neurons in the spinal cord. In this case the lumbar enlargement extract after its intracisternal injection to healthy animals increases the duration of hind limb passive extension on the side where the donor neurons are hyperactive. The extract of the spinal cord of healthy rats was ineffective. Proteolysis of the extract with pronase or co-injection of opiate antagonist--naloxone--completely eliminated the lateralized changes in the muscular tone of the recipient. Substances that cause the unilateral changes in the muscular tone of the recipient are believed to be peptides. They are assumed to be involved in the functioning of endogenous opioid system. The level of substance P in the donor spinal cord was elevated bilaterally, but was higher in the hyperactive half of the spinal cord.