Effects of phosphocreatine on the heart nerve structures under conditions of local immune impairment (electrophysiological and neurochemical aspects)

Effects of phosphocreatine on the neurohumoral mechanisms controlling the heart under conditions of a local immune injury were studied in acute experiments on anesthetized dogs using electrophysiological, biochemical, and electron microscopy techniques. After the development of heart injury, cardiogenic depressor reflexes evoked by an intracoronary injection of veratrine and mediated by vagus mechanisms disappeared, while pressor reflexes became dominating. This phenomenon correlated with an increase in concentration of a vasoconstrictor agent, leucotriene LTC₄, in the blood and with a considerable ultrastructural impairment of nerve terminals in the myocardium. Preliminary injection of phosphocreatine prevented the development of structural impairments, favored the preservation of vagosympathetic depressor reflex, and not only prevented the increase in LTC₄ concentration, but even dramatically decreased its level (by 82%, as compared with the initial level). We concluded that complex protective effect of phosphocreatine provides structural and functional preservation of the receptor apparatus in the heart and can play a considerable role in normalization of neurohumoral mechanisms controlling the heart under conditions of pathological impairment.Neirofiziologiya/Neurophysiology, Vol. 27, No. 2, pp. 140–146, March–April, 1995.     

Effect of phosphocreatine and related compounds on the phospholipid metabolism of ischemic heart

Changes in the content of lysophosphoglycerides in a crude plasmalemmal fraction of canine heart during short-term ischemia (occlusion of the left descending coronary artery for 8 min) have been studied in the presence and in the absence of phosphocreatine and phosphocreatinine. In the control experiments without PCr or PCr-nine ischemia caused significant elevation of the content of LPG: that of lysophosphatidylcholine was increased by 83% and that of lysophosphatidylethanolamine by 168%. Intravenous administration of PCr and PCr-nine in doses of 300 mg/kg completely prevented accumulation of LPG in the ischemic zone. Because of the well-known arrhythmogenic properties of LPG, the inhibitory effect of PCr and PCr-nine on the elevation of their concentration in the ischemic zone may be closely related to the antiarrhythmic action of PCr and PCr-nine in acute myocardial ischemia.     

Effect of phosphocreatine on reflex activity of the brain stem reticular formation

The effects of phosphocreatine (PCr) on startle reflexes, known to be relayed through the brain stem reticular formation (RF), were investigated on chloralose-anesthetized rats. PCr (10^–6 to 10^–3 M) was either applied superficially to the bottom of the fourth ventricle or microinjected into the reticular gigantocellular nucleus. The PCr effect was found to depend on its concentration. At low concentrations (10^–6 to 10^–5 M), PCr markedly facilitated the reflexes; sometimes its application gave rise to additional, later and longer, discharges following startle reflexes, whereas an inhibitory effect predominated at higher PCr concentration (10^–4 to 10^–3 M). Possible mechanisms of PCr action on the studied reflexes are discussed.Neirofiziologiya/Neurophysiology, Vol. 25, No. 3, pp. 272–278, 1993.     

Sympathetic neural influence on bone metabolism in microgravity (Review)

Bone loss is one of the most important complications for astronauts who are exposed to long-term microgravity in space and also for bedridden elderly people. Recent studies have indicated that the sympathetic nervous system plays a role in bone metabolism. This paper reviews findings concerning with sympathetic influences on bone metabolism to hypothesize the mechanism how sympathetic neural functions are related to bone loss in microgravity. Animal studies have suggested that leptin stimulates hypothalamus increasing sympathetic outflow to bone and enhances bone resorption through noradrenaline and β-adrenoreceptors in bone. In humans, even though there have been some controversial findings, use of β-adrenoblockers has been reported to be beneficial for prevention of osteoporosis and bone fracture. On the other hand, microneurographically-recorded sympathetic nerve activity was enhanced by exposure to microgravity in space as well as dry immersion or long-term bed rest to simulate microgravity. The same sympathetic activity became higher in elderly people whose bone mass becomes generally reduced. Our recent findings indicated a significant correlation between muscle sympathetic nerve activity and urinary deoxypyridinoline as a specific marker measuring bone resorption. Based on these findings we would like to propose a following hypothesis concerning the sympathetic involvement in the mechanism of bone loss in microgravity: An exposure to prolonged microgravity may enhance sympathetic neural traffic not only to muscle but also to bone. This sympathetic enhancement increases plasma noradrenaline level and inhibits osteogenesis and facilitates bone resorption through β-adrenoreceptors in bone to facilitate bone resorption to reduce bone mass. The use of β-adrenoblockers to prevent bone loss in microgravity may be reasonable.     

Thyrotropin-releasing hormone (TRH)-immunoreactive structures in the brain of the domestic mallard

Summary The distribution of immunoreactive thyrotropin-releasing hormone (TRH) in the central nervous system of the domestic mallard was studied by means of the peroxidase-antiperoxidase technique. After colchicine pretreatment, the highest number of TRH-immunoreactive perikarya was found in the parvocellular subdivision of the paraventricular nucleus and in the preoptic region; a smaller number of immunostained perikarya was observed in the lateral hypothalamic area and in the posterior medial hypothalamic nucleus. TRH-immunoreactive nerve fibers were detected throughout the hypothalamus, forming a dense network in the periventricular area, paraventricular nucleus, preoptic-suprachiasmatic region, and baso-lateral hypothalamic area. TRH-containing nerve fibers and terminals occurred in the organon vasculosum of the lamina terminalis and in the external zone of the median eminence in juxtaposition with hypophyseal portal vessels. Scattered fibers were also seen in the internal zone of the median eminence and in the rostral portion of the neural lobe. Numerous TRH-immunoreactive fibers were detected in extra-hypothalamic brain regions: the highest number of immunoreactive nerve fibers was found in the lateral septum, nucleus accumbens, olfactory tubercle, and parolfactory lobe. Moderate numbers of fibers were located in the basal forebrain, dorsomedial thalamic nuclei, hippocampus, interpeduncular nucleus, and the central gray of the mesencephalon. The present findings suggest that TRH may be involved in hypophysiotropic regulatory mechanisms and, in addition, may also act as neuromodulator or neurotransmitter in other regions of the avian brain.     

Protective effect of peptide semax (ACTH(4-7)Pro-Gly-Pro) on the rat heart rate after myocardial infarction

Semax, a member of ACTH-derived peptides family, was used in treatment of ischemic stroke in patients. It decreased neurological deficiency and reduced NO hyperproduction in the rat brain caused by acute cerebral hypoperfusion. We suggest that semax is also capable of protecting the rat heart from ischemic damage 28 days after myocardial infarction (MI) induced by left descendent coronary artery occlusion. Semax (150 microg/kg) was given i. p. in the operating day twice: 15 min and 2 hours after coronary occlusion, and once a day for the following 6 days. In 28 days after infarction, the MI group developed cardiac hypertrophy, cell growth was caused mainly by the increase of contractile filaments not supported by the appropriate mitochondrial growth that indicated an impaired energy supply of the cells. Moreover, cardiac hypertrophy was accompanied by decreased mean arterial blood pressure and cardiac contractile function and increased left ventricular end-diastolic pressure. Pharmacological change of cardiac afterload revealed that, in 28 days after MI, the rat heart was not able to change its contractile performance in response to either increase or decrease of systemic blood pressure, and as a result could not maintain its diastolic pressure. All these changes obviously reflect development of heart failure. Semax did not affect cardiac work but partially prevented end-diastolic pressure growth in left ventricle as well as ameliorated cardiomyocyte hypertrophy and disproportionate growth of contractile and mitochondrial apparatus, thus exerting beneficial effect on the left ventricular remodeling and heart failure development late after myocardial infarction.     

In vivo development of brain phosphocreatine in normal and creatine-treated rabbit pups

To study the effects of creatine (Cr) on brain energy metabolism and on hypoxia-induced seizures, 5- to 30-day-old rabbit pups were given subcutaneous Cr (3 g/kg) for 3 days before exposure to 4% O2 for 8 min. In saline-treated controls, hypoxic seizures were most frequent at 15 days (80% of pups) and 20 days (60%) of age. Seizures were prevented at 15 days and reduced 60% at 20 days in Cr-treated pups. In surface coil-localized brain 31P nuclear magnetic resonance spectra, with signal from both cerebral gray (GM) and white (WM) matter, the phosphocreatine (PCr)/nucleoside triphosphate (NTP) ratio doubled between 5 and 30 days of age in controls. In all Cr-injected pups, brain PCr/NTP increased to values seen in 30-day-old controls. When spectra were acquired in predominantly GM and WM slices in vivo, the PCr/NTP ratio was very low in GM at 5 days but reached adult levels by 15 days in controls. In WM, the ratio increased steadily from 5 to 30 days of age. In Cr-injected pups, PCr/NTP increased to mature levels in WM and in GM at all ages. In conclusion, hypoxic seizures occur midway in the time course of brain PCr/NTP increase in rabbit pups as previously described in rat pups. In both altricial pups, systemic Cr increases brain PCr/NTP ratio and prevents hypoxic seizures. These results suggest that mature levels of PCr and/or Cr in brain limit EEG activation either directly or indirectly by preventing hypoxic metabolic changes.     

The influence of neuromodulators on the synaptic plasticity in dopaminergic structures of the midbrain (hypothetical mechanism)

A mechanism underlying the effects of neuromodulators on long-term changes in the efficacy of excitatory and inhibitory inputs to dopaminergic and inhibitory cells of the substantia nigra and ventral tegmental area is suggested. According to this mechanism, activation of Gi/0 protein-coupled dopamine D2 autoreceptors and opioid kappa (mu) receptors on dopaminergic (inhibitory) cells promotes the LTD of excitatory inputs to these cells and decrease in their activity. Activation of Gq/11 protein-coupled alpha1 adrenoreceptors, muscarinic M1, neurokinin NK3 (alpha1, M3, NK1, serotonin 5-HT2) receptors on dopaminergic (inhibitory) cells as well as activation of Gs protein-coupled D1 receptors on inhibitory cells promotes the LTP of excitatory inputs to these cells and increase in their activity. Augmenting (lowering) GABA release can be provided by activation of presynaptic D1 and M3 receptors (mu, 5-HT1, and adenosine A1) receptors. Increase (decrease) in GABA concentration due to modulation of inhibitory cell activity and/or GABA release will promote the induction of LTD (LTP) of excitatory inputs to target dopamine cells. The model agree with known experimental data describing the involvement of neuromodulators in modification of dopamine cell activity and dopamine release. The suggested model can be useful in understanding the operation of neuronal networks, which include the basal ganglia.     

An (1)H-MRS evaluation of the phosphocreatine/creatine pool (tCr) in human muscle

The human gastrocnemius was examined with and without creatine supplementation under the conditions of rest, ischemic fatigue (IF), and recovery to perturb the pool sizes and equilibrium between phosphocreatine (PCr) and creatine (Cr). (1)H- and (31)P-magnetic resonance spectroscopy (MRS) were used to examine the total creatine (tCr) pool in each of the metabolic states. (31)P-MRS monitored the depletion of the PCr peak during IF to <5% of that at rest. (1)H-MRS focused on the tCr methyl peak at 3.02 ppm (dipolar coupled triplet), at which point it was expected that the triplet peak intensity would be similar both in IF and rest. Initial (1)H-MRS data showed the peak intensity during IF decreased, suggesting a change in tCr pool size. Subsequent studies of transverse relaxation time (T(2)) revealed that this decline was primarily due to a more rapid T(2) decay of the tCr peak in IF (T(2) approximately 40 ms) compared with at rest (T(2) approximately 162 ms). Because Cr is the major contributor to tCr in IF, it is possible that there is a pool of Cr displaying reduced mobility in vivo. Moreover, the residual dipolar coupled triplet observed at rest collapsed into a broad singlet during IF, suggestive of significant changes in the ordered environment experienced at rest for PCr compared with when it is converted to Cr during IF. In addition, these data suggest that in (1)H-MRS studies whose goals include quantitative estimates of tCr pool sizes, standardized metabolic conditions or careful T(2) evaluations will be required.     

Changes in the functional parameters of the brain structures under the influence of enkephalin-like tetrapeptidamide

In complex neurophysiological and cytobiochemical study single injections of tetrapeptide amide (TPA) caused a short-term analgetic effect which manifested itself in the absence of motor reactions and EEG changes of cortical and subcortical brain structures after painful stimulation of extremities. This effect was accompanied by changes of some indices of transmitter (monoamine oxidase) and protein metabolism in the cerebral hemispheres at cellular and subcellular levels. In 30-40 min after a TPA injection, EEG suppression and absence of EPs to light flashes were observed in cortical and subcortical structures. Simultaneously motor disorders developed. The observed EEG changes had an undulatory character: on the second day EEGs were restored and on the third day--suppressed once again. This period of TPA action was accompanied by varied changes of the investigated types of metabolism. The question of the necessity of systemic approach to the study of TPA action is discussed, as such an approach allows to reveal complex neurophysiological and fine biochemical relations in the reactions of brain structures and in animal behaviour.     

The influence of hypothermia on the restoration of neural activity of brain slices from hibernating ground squirrels

1. 1. Neural activity was recorded in hippocampal slices from deep hibernating Yakut ground squirrels and in hippocampal and septal slices from non-hibernating animals.

2. 2. Slices were placed immediately after preparation in hypothermic conditions (3–4°C). Their activity was tested under standard conditions at 31°C in the incubation chamber. Some of the prepared slices were tested after maintenance in hypothermia for 2 or 24 h.

3. 3. In the hippocampal slices of hibernating ground squirrels, neural activity was present, irrespective of the period in hypothermia.

4. 4. Slices from guinea-pigs and hamsters did not possess neural activity after either 2 or 24 h of hypothermic treatment.

     

Atrial natriuretic factor (ANF) binding sites in brain and related structures

Visualization of [¹²⁵I]ANF binding sites in rat brain by an autoradiographic technique demonstrated that these sites are highly localized in areas such as the olfactory bulb, subfornical organ, area postrema and nucleus tractus solitarius. This distribution suggests that certain cardiovascular effects of ANF could be centrally mediated and that the existence of brain ANF-related peptides should be considered. Finally, moderate densities of [¹²⁵I]ANF binding sites are found in the rat and guinea pig eye while low densities are seen in pituitary and pineal gland.     

The influence of phentolamine on the fluorescence of catecholaminergic structures in selected areas of rat brain.

Wistar rats were given phentolamine into the ventriculus lateralis. The D1 group of rats received a larger dose than the D2 group. The animals were decapitated within 2 hours after phentolamine injection. The FALCK fluorescence technique was applied to demonstrate the fluorescence of catecholaminergic structures. The results in rat brain areas selected according to KONIG and KLIPPEL are shown in figs 10-46. Whereas earlier biochemical experiments did not show any phentolamine-induced changes in the NA level of the whole brain, the present histochemical experiments carried out with the fluorescence technique revealed the influence of phentolamine on individual structures and areas of the NA system. In comparison with the control material in 5 out of 11 areas the fluorescence was much weaker, in 3 it was similar to the control group, and in 3 (1 with a larger dose and 2 with a smaller dose of phentolamine) it was slightly stronger. The simultaneous increase of fluorescence in 6 out of 7 areas of the DA system may indicate a compensatory interaction of these areas as a response to the neuromediator decrease in the NA system. The paper discusses the increase in the intensity of fluorescence induced by a small dose of phentolamine in some brain areas, or by a large dose in others, both these alternatives depending on the neuromediator turnover.