Changes in energy metabolism in several brain regions and vegetative responses of white rats during neurotization

Changes of vegetative reactions and cytochrome oxidase (CChO) activity in various brain structures were studied in rats during neurotization. One week neurotization led to an increase of arterial blood pressure, respiration rate, cardiac stroke volume and heart rate. In three weeks of neurotization there was a decrease of stroke volume accompanied by an increase of heart rate and some decrease or respiration rate leading to a reduction of oxygen consumption. Neurotization during one and especially three weeks elicited an enhancement of CChO activity in various brain areas, more pronounced in the cerebral cortex. A four week "rest" after neurotization during three weeks normalized the CChO activity. CChO activation during neurotization is supposed to be one of the mechanisms of adaptation to hypoxia accompanying neurosis.     

Changes in the cytochrone C and A (A3) content in the liver mitochondria of rats with hyperthyroidism

Thyroxin was administered daily to male Wistar rats aged 1, 3, 12 and 24 months in a dose of 250 g per 100 g body weight. After 9 days there was a significant increase in the content of cytochromes c and a (a(3)). An appreciable rise in the cytochrome c content was recorded as long as after one day, whereas the content of cytochromes a (a(3)) did not exceed normal even after 2 days. Following one day the content of cytochromes a (a(3)) in 3-, 12-, and 24-month-old rats was sightly lower than in normal rats. A significant temporary increase in the c/a (a(3)) ratio was observed after 1-2 days only in 12- and 24-month-old rats. The c/a (a(3)) ratio increased with age. It is suggested that application of thyroid hormones may be promising in studies on the regulation of mitochondrial biogenesis.     

Effect of pyridoxal phosphate on gamma-aminobutyric acid metabolism in different sections of the brain in irradiated animals]

A study was made of the effect of X-rays (4,5 Gy) and pyridoxal phosphate (3 mg/kg, v/v) on the activity of pyridoxal enzymes of GABA metabolism (e.g. glutamate decarboxylase, E.C. 4.1.1.15) and aminobutyrate aminotransferase (GABA-T, E.C. 2.6.1.19), as well as on GABA and glutamate content of the hemisphere cortex, brain stem and cerebellum of rabbits 6 and 10 days following irradiation and injection of a coenzyme. The height of the radiation sickness in rabbits was characterized by the manifest changes in glutamate decarboxylase and GABA-T activity, as well as in GABA and glutamate content of various brain parts differing in the structural and functional functions. The administration of pyridoxal phosphate produced pronounced activation of glutamate decarboxylase, particularly 6 days after irradiation and administration of the co-enzyme, and, to a lesser extent, influenced GABA-T function. Pyridoxal phosphate favored maintaining the GABA level above the control level in the hemisphere cortex and brain stem 6 and 10 days after exposure. The injection of pyridoxal phosphate did not normalize the glutamate content of the brain parts 6 days after exposure, but favored the normalization of GABA-T activity on day 10.     

Effect of thyroidectomy on phosphorylative oxidation and RNA synthesis in various regions of the brain in the adult monkey

The subcellular effects of thyroidectomy in selected brain regions of Cynomolgus monkey were analyzed. 20 days after operation the respiratory rates, the activities of succinate cytochrome c reductase, glycerol-3-phosphate dehydrogenase and of oligomycin-sensitive ATPase were decreased in mitochondria isolated from all brain structures. The highest reduction (30%) was found in cerebral cortex and hippocampus. Cerebellar and striatal activities were reduced by about 20%. A smaller decrease (15%) was observed in thalamus. The effects of thyroidectomy on in vitro RNA synthesis were followed in cerebral cortex, cerebellum and thalamus. In the three analyzed regions, the activities of nucleolar and nucleoplasmic RNA polymerases dropped by 40%. Replacement therapy with T4 (2.5 micrograms/kg/day) or T3 (1 microgram/kg/day) administered immediately after thyroidectomy for 20 days, maintained mitochondrial and nuclear activities at normal level.     

Total RNA content and blood flow in rat brain after RNA administration

The changes in blood flow through selected brain structures and the changes in the total RNA content of cells of these structures were examined after a single administration of yeast RNA to 6-month-old male rats. The total content of ribosomal RNA in cells of the limbic system (septum, hippocampus, hypothalamus) increased 48 hrs after the administration of 100 mg i.p. yeast RNA , dropped after 7 days (in hypothalamus), 21 and 30 days (in hippocampus), 30 days (in septum). In cells of the limbic system as a whole there is a higher total RNA content in experimental rats. No changes were observed in the cells of parietal brain cortex. Blood flow increased in limbic structures 21 and 30 days after RNA administration and in septum and in hippocampus also 90 days after application. No changes were observed in parietal brain cortex, bulbi olfactorii, cerebellum and brain stem. Histochemical changes correlated positively with blood flow changes in the limbic system 14, 21, 30 and 90 days after RNA application. The body weight of experimental rats did not differ from that of control animals. The changes in haemodynamic parameters were transient and were demonstrated as fluctuations in heart rate, cardiac output, and peripheral resistance. Blood pressure experienced no changes.     

Postnatal development of energy metabolism in the rat brain

The activities of cytochrome c oxidase and F0F1-ATPase as well as the content of cytochromes cc1, aa3, and b were investigated in free brain mitochondria in the course of postnatal development and aging. The results show an increase of Vmax of both enzymes during postnatal development (between day 5 and 30). During the following phase ending at the age of 6 months, a decrease of F0F1-ATPase and cytochrome c oxidase activity occurs. From 6 to 12 months of age the activity of these enzymes did not change. The KM for both enzymes remained unchanged during the whole period observed. The content of cytochromes increased from the low values found in young rats, reached the highest values at around one month, and decreased till the age of 3 months. Later, their content in brain mitochondria did not markedly change. Our results suggest that the metabolic maturation of brain mitochondria differs in several aspects from the same process in other tissues, mainly in the time course. This is probably due to the unique role of neural tissue in the organism.     

Changes in the concentrations of total RNA and DNA in the cerebral cortex and internal organs (liver and heart) after prolonged hypovolemic hypotension and in the post-resuscitation period

It was shown in experiments on dogs that after 4-hour hypovolemic hypotension the content of total RNA in brain cortex and myocardium homogenates decreased. In the liver, there was a significant decrease both in RNA and DNA content. In the postresuscitation period, the content of nucleic acids in the myocardium returned to normal after 14--21 days, and that in the liver after 3--4 months. The gray matter of the brain manifested a delayed lowering of DNA content (after 14--21 days), and the level of nucleic acid did not return ot normal over 3--4 months after resuscitation.     

Postnatal development of the scratch reflex in rabbits]

The development of the scratch reflex was studied in newborn (up to 2 months old) rabbits in norm and after elimination or activation of some parts of their nervous system (reticular formation, cerebellum, caudate nucleus, cerebral cortex, superior cervical sympathetic ganglia). The experiments with the section of the brain stem at the border between the medulla and the midbrain showed that in very young (5-10 days old) rabbits in norm the scratch reflex is controlled by the spinal cord with no influences of structures situated above the section's level. Later on the spinal mechanism of the scratch reflex becomes subject to supraspinal influences, among which in 2-3 weeks old animals facilitatory effects are predominant produced, in particular, by the reticular formation and the cerebellum, whereas in older age prevail inhibitory influences of the cerebral cortex, cerebellum, caudate nucleus and the sympathetic nervous system.     

Distribution and activity of pyridoxal kinase in human brain during ontogenesis]

The activity of pyridoxal kinase was sharply increased in whole brain tissue of human, embryos and fetuses within 6-11 weeks of development. In brain stem the maximal values of the enzyme activity was observed at early stages of prenatal development of fetuses. The activity of pyridoxal kinase was increased in cerebral cortex and in the limbic system up to complete maturation of fetuses. It correlated with the fetus age within 14-40 weeks of development as calculated per 1 g of tissue wight or 1 mg of protein. The enzyme is distributed evenly in brain of newborns, babies and adult people. Its activity in grey cortex substance is higher, than in white one. There are 2-10-fold individual fluctuations of pyridoxal kinase activity in brain of people without CNS pathology. In newborns, having prolonged hypoxy at prenatal period, the enzyme activity was on the average by 70-80% lower at different brain parts than in newborns which had no primary asphyxia. A low pyridoxal kinase activity (not more than 1-5% as compared with its normal level) was observed in different brain parts of a child affected by focal gliosis and epilepsy.     

Analysis of the level of cyclic adenosine-3',5'-monophosphate in structures of the 'informational' and 'motivational' system of the rat brain during active conditioning

The content of cyclic adenosine-3',5'-monophosphate (cAMP) was studied in structures of the "motivational" and "infromational" systems of rat brain after the active avoidance conditioning procedure in rats. Three groups of animals were examined: naive rats, trained (conditioned) rats, and group of the active control presented with uncombined conditioned (light) and unconditioned (electric footshock) stimuli. The content of cAMP was determined in the frontal cortex, hippocampus, amygdala, and hypothalamus of both hemispheres immediately after the retrieval of conditioned reaction one day after conditioning. A significant increase in cAMP level was bilaterally observed in the hypothalamus in the group of active control, and in both hippocampi and the right frontal cortex in the conditioned animals. Positive correlations between the cAMP levels in symmetrical regions of the frontal cortex, amygdala, and hypothalamus were revealed in all the examined groups. Additionally, intra- and interhemispheric correlations were found in the active control and conditioned rats. Patterns of correlation were specific for each of these groups. The observed phenomenon is discussed in term of involvement of "informational" and "motivational" brain structures in the mechanisms of adaptive behavior.     

THE EFFECTS OF REPEATED-LYSERGIC ACID DIETHYLAMIDE INJECTIONS ON CATECHOLAMINE LEVELS AND TYROSINE HYDROXYLASE ACTIVITY IN RAT BRAIN REGIONS

Daily injections of 100 μg/kg of d -lysergic acid diethylamide (LSD) for 14 days produced a significant decrease in the dopamine level in rat brain corpus striatum which was still apparent 15 days after the last LSD treatment. Further LSD injections did not change the amount of dopamine depletion. In cerebral cortex, 14 days of LSD injections produced a significant decrease in the norepinephrine level and a significant increase in tyrosine hydroxylase activity. The elevated tyrosine hydroxylase activity was still present 15 days after the final LSD injection but only in those animals receiving daily vehicle injections during this period. Pre-treatment of rats with daily saline injections for 2 weeks before the 2 week period of LSD treatment prevented both the reduced norepinephrine content and elevated tyrosine hydroxylase activity usually found 24 h after the last LSD injection.     

Differential time course of induction of rat liver microsomal cytochrome P-450 isozymes and epoxide hydrolase by Aroclor 1254

The time course of induction of rat liver microsomal cytochromes P-450a, P-450b + P-450e, P-450c, and P-450d and epoxide hydrolase has been determined in immature male rats administered a single large dose [1500 mumol (500 mg)/kg body wt] of the polychlorinated biphenyl mixture Aroclor 1254. Differential regulation of these xenobiotic-metabolizing enzymes was indicated by their characteristic patterns of induction. The rate of induction of cytochrome P-450a and epoxide hydrolase was relatively slow, and steady-state levels of these enzymes were maintained from approximately Days 9 to 15 after Aroclor 1254 treatment. In contrast, cytochrome P-450c was maximally induced 2 days after Aroclor 1254 treatment and remained at a constant level through Day 15. Steady-state levels of cytochrome P-450d, beginning 1 week after Aroclor 1254 treatment, were preceded by a fairly rapid rate of induction and possibly by a small decline from maximal levels observed around Days 4 to 5. Like those of the other cytochrome P-450 isozymes and epoxide hydrolase, the levels of cytochromes P-450b + P-450e were constant from Day 9 to 15 after Aroclor 1254 treatment. However, an unexpected but reproducible decline (approximately 25%) in total cytochrome P-450 content observed between Days 4 and 9 after Aroclor 1254 treatment principally reflected a dramatic and totally unanticipated decrease (approximately 45%) in the level of cytochromes P-450b + P-450e. This transient decline in the level of cytochromes P-450b + P-450e was not due to an unusual effect of a mixture of polychlorinated biphenyls, since identical results were obtained with two individual congeners, namely 2,3,4,5,4'-penta- and 2,3,4,5,3',4'-hexachlorobiphenyl, that induced the same isozymes as Aroclor 1254. In contrast, when rats were treated with 2,4,5,2',4',5'-hexachlorobiphenyl, which induces cytochromes P-450a and P-450b + P-450e and epoxide hydrolase but not cytochromes P-450c or P-450d, maximal levels of cytochromes P-450b + P-450e were attained on Day 4 and no decrease was observed over the next 11 days. These results suggest that there may be an interaction in the regulation of induction of certain individual cytochrome P-450 isozymes.     

Interhemispheric asymmetry in the functional organization of the prefrontal cortex in animals of different typologic groups

Rats allocated to groups by the method of "emotional resonance": rats which did and did not escape crying of a partner (A- and E-groups, respectively). Unit activity in the right and left prefrontal brain cortex (PFC) was recorded in these rats. The recorded neurons neurons were divided in two groups according to their reaction to a change in the level of food motivation. The so-called D-neurons decreased their activity after feeding of animals after a 24-hour food deprivation and the other group (I-neurons) increased its firing rate rate in this situation. It was shown that hemispheric distributions of D- and I-neurons are different in selected rat groups. In E-rats the I-neurons substantially predominated in the left hemisphere, whereas the D-neurons were more frequently recorded in the right one. No such asymmetry was observed in A-group of rats. During intracranial stimulation of emotionally positive brain structures I-neurons increased their firing rate, predominantly, in the left hemisphere, whereas during intracranial emotionally negative stimulation activation of the D-neurons predominated at the right. Features of the observed functional interhemispheric asymmetry of prefrontal cortex in A- and E-groups of rats were explained by differences in the interaction between hemispheres and dissimilar activation control.     

Therapeutic doses of SkQ1 do not induce cytochromes P450 in rat liver

The effect of SkQ1 (a mitochondria-targeted antioxidant) on the level of cytochromes P450 in rat liver was studied. It was found that administration of therapeutic dose of SkQ1 with drinking water for 5 days (250 nmol/kg of body weight per day) did not alter the level of cytochromes P450. Under the same conditions, the standard dose of phenobarbital used for the induction of cytochromes P450 caused the 2.7-fold increase in the content of these cytochromes. We conclude that therapeutic doses of SkQ1 do not induce cytochromes P450 in rats.     

Unilateral microdialysis of gabazine in the dorsal medulla reverses thermal prolongation of the laryngeal chemoreflex in decerebrate piglets.

The laryngeal chemoreflex (LCR) is elicited by water in the larynx and leads to apnea and respiratory disruption in immature animals. The LCR is exaggerated by the elevation of brain temperature within or near the nucleus of the solitary tract (NTS) in decerebrate piglets. Thermal prolongation of reflex apnea elicited by superior laryngeal nerve stimulation is reduced by systemic administration of GABA(A) receptor antagonists. Therefore, we tested the hypothesis that microdialysis within or near the NTS of gabazine, a GABA(A) receptor antagonist, would reverse thermal prolongation of the LCR. We examined this hypothesis in 21 decerebrate piglets (age 3-13 days). We elicited the LCR by injecting 0.1 ml of water into the larynx before and after each piglet's body temperature was elevated by approximately 2.5 degrees C and before and after 2-5 mM gabazine was dialyzed unilaterally and focally in the medulla. Elevated body temperature failed to prolong the LCR in one piglet, which was excluded from analysis. Elevated body temperature prolonged the LCR in all the remaining animals, and dialysis of gabazine into the region near the NTS (n = 10) reversed the thermal prolongation of the LCR even though body temperature remained elevated. Dialysis of gabazine in other medullary sites (n = 10) did not reverse thermal prolongation of the LCR. Gabazine had no consistent effect on baseline respiratory activity during hyperthermia. These findings are consistent with the hypothesis that hyperthermia activates GABAergic mechanisms in or near the NTS that are necessary for the thermal prolongation of the LCR.     

BRAIN DOPAMINE-β-HYDROXYLASE: REGIONAL DISTRIBUTION AND EFFECTS OF LESIONS AND 6-HYDROXY-DOPAMINE ON ACTIVITY

Abstract— A modification of a specific and sensitive radioassay was used to measure dopamine-β-hydroxylase (DBH) (EC 1.14.2.1) in various regions of the rat CNS. Highest activity was found in the hypothalamus. Relative to activity in the hypothalamus (= 100 per cent), activity in brainstem was 80 per cent, in sensory motor cortex 55 per cent, in caudate nucleus 32 per cent, and in cervical spinal cord 30 per cent. Two to three weeks after a unilateral electrolytic lesion of the lateral hypothalamus, activity of DBH in the ipsilateral cerebral cortex fell to 17 per cent of control values without changes in activity ipsi- or contra-laterally in the brainstem. Thalamic lesions did not affect DBH activity. In cerebral cortex contralateral to the hypothalamic lesion, enzymic activity rose 30 per cent. After intracisternal administration of 6-hydroxy-dopamine (6-OH-DA), cortical DBH activity fell to 20 per cent of control values. Reserpine (3 mg/kg subcutaneously for 3 days) did not increase the activity of DBH in brain regions but did increase the activity of DBH in adrenal gland 200 per cent. Our results suggest that: (a) DBH is widely distributed in neurons in CNS with a regional pattern of activities that appears to parallel the Jevels of norepinephrine; (b) DBH activity in the cerebral cortex depends on the integrity of structures (e.g. medial forebrain bundle) in lateral hypothalamus; (c) DBH in brain areas lacking cell bodies of nore- pinephrine-neurons (e.g. cerebral cortex) is contained in norepinephrine-containing axon terminals and (d) the activity of DBH in brain is not increased by reserpine under conditions that provoke marked increase of DBH activity in the adrenal gland.     

Cyclic adenosine-3',5'-monophosphate content in the limbic system structures of the rat brain on the administration of corticosteroids

The effect of hydrocortisone and DOCA on the cAMP content in the hypothalamus, hippocampus and striate body of the rat brain was investigated. Single (determined after 1 and 24 hours) and repeated (7 days) hydrocortison administration in a dose of 5 mg/100 g body weight was accompanied by an increase in the cAMP concentration in the brain structures under study. Single administration of DOCA in a dose of 0.5 mg/100 g body weight did not produce any changes in the cAMP level in the structures of the rat brain limbic system; however, the dose of 2.5 mg raised the cAMP level. Prolonged administration of the hormone in the above doses dod not change the cAMP level in the brain structures. Only the hippocampus showed a 210% increase in the cAMP level during DOCA administration in a dose of 0.5 mg.     

Level of neuroactive steroids in the brain and sex-related peculiarities of formation and extinction of conditioned reflex in rats

Sex-related peculiarities of dynamics of brain sex steroids in the process of learning and extinction of the conditioned reflex of passive avoidance have been studied in model experiment. Prior to learning of the conditioned reflex, female rats were found to be distinguished by manifestation of anxiety and fear as compared with male rats. At formation of the conditioned reflex, no significant sex-related differences were detected between males and females, whereas extinction of the conditioned reaction of passive avoidance in males occurred by 2–3 days faster than in females. At learning of conditioned reaction of passive avoidance, in sexually mature male rats there was revealed an increase of the testosterone content in various brain structures, especially in hippocampus and frontal cortex, while its level in blood plasma remained unchanged. Also shown was an elevation of estradiol concentration in female amygdale, whereas at extinction of the conditioned reaction of passive avoidance, a rise of estradiol values was noted in hippocampus and cingular cortex. At the same time, the testosterone level in blood plasma did not change, whereas after extinction of the conditioned reflex the estradiol concentration decreased statistically significantly. Different dynamics of changes of the sex steroid levels in brain and blood plasma can indicate a possibility of their formation in the nervous tissue. The performed correlation analysis confirms the concept of selective involvement of testosterone and estradiol of individual brain structures in realization of processes of learning and memory in sexually mature male and female rats.     

Terminal Electron Transport in Leptospira

The cytochrome content of three leptospiral strains grown in several media was investigated after it was shown that respiratory inhibitors suppressed oxygen consumption of a water isolate, B(16), and that two pathogenic serotypes, pomona and schueffneri, were active catalase producers, whereas B(16) lacked catalase activity. Reduced minus oxidized difference spectra disclosed cytochromes of the a, c, and c(1) types in all strains. Although no spectral evidence suggested the existence of cytochrome b components, they could have been masked by cytochrome c, and their presence cannot be ruled out. Carbon monoxide difference spectra revealed peaks indicative of a cytochrome oxidase of the o type in all strains. Carbon monoxide spectra further suggested that a cytochrome a oxidase, possibly a(1) or a(3), and a pigment with absorption spectra different from those of previously characterized cytochromes existed in the two pathogens and not in the water isolate. Physiological reduction of the cytochromes by various metabolic substrates implied that the cytochrome system in Leptospira was functional. No effect of the various growth media on the cytochrome patterns of the three strains was observed, indicating that all three strains were capable of synthesis of cytochrome components and did not require heme prosthetic groups present in serum.     

Studies on the effect of iron overload on rat cortex synaptosomal membranes

Iron as ferrous ammonium sulfate was injected into the cerebral spinal fluid of rats. After three consecutive days of injection of 4 mumol of iron, the total iron content of brain cortex synaptosomes from the iron-treated animals was 2-fold higher than that from control animals receiving the saline vehicle only. Spin label studies of the synaptosomal membranes demonstrated that the lipid region of the membranes became more rigid and, in addition, the mobility of labeled SH groups of membrane proteins decreased after the iron treatment. The cholesterol content was significantly higher in iron-treated animals as compared to controls. Centrophenoxine pretreatment (100 mg/kg body weight daily for 6 weeks) diminished the iron effects. Synaptosomal membrane alterations observed after iron treatment were similar to changes observed previously during aging. This lends support to the notion that free-radical induced damage occurs in brain membranes with increasing age.