The aim of this study was to examine whether threshold to heat stimuli, and expression of transient receptor potential vanilloid1 (TRPV1) and nerve growth factor (NGF) in dorsal root ganglion (DRG) altered under conditions of long-term limb immobilization. A plastic cast was wrapped around the right limb from the forearm to the forepaw to keep wrist joint at 90° of flexion for 5 weeks. Heat hyperalgesia was tested using the plantar test at 6 h after removing cast. The rats were perfused transcardially with 4 % paraformaldehyde and DRGs were excised at 24 h after removing cast. For size distributions of the TRPV1-IR and NGF-IR neuronal profile, the DRG area measurements over 1000 DRG neurons per animal were measured in each side, on both the immobilized (ipsilateral) and contralateral sides. Ipsilateral withdrawal latency was significantly shorter than contralateral sides. Ipsilateral percentage of immunoreactive neurons in the total DRG neurons was significantly higher than contralateral sides in TRPV1-IR and NGF-IR. Long-term casting induced heat hyperalgesia, and up-regulation and phenotypic change of TRPV1-IR and NGF-IR in DRGs on the immobilized side. These DRG alterations may involve heat hyperalgesia after long-term limb immobilization., T. Nishigami, ... [et al.]., and Obsahuje seznam literatury
The hypothalamic supraoptic and paraventricular nuclei consist of oxytocin and arginine vasopressin synthesizing neurons that send projections to the neurohypophysis. A growing body of evidence in adult animals and young animals at near term confirmed the structure and function in the vasopressinergic and oxytocinergic network. However, whether those distinctive neural networks are formed before near term is largely unknown. This study determined the special patterns in location and distribution of oxytocin- and vasopressin-neurons in the paraventricular and supraoptic nuclei from preterm to term in the ovine fetuses. The results showed that oxytocin- and vasopressin-neurons were present in both nuclei at the three gestational time periods (preterm, near term, and term). In the paraventricular nuclei, vasopressin-cells concentrated mainly in the core of the middle magnocellular paraventricular nuclei, and oxytocin-cells were scattered surrounding the core. In the supraoptic nuclei, vasopressin-cells mostly located in the ventral part, and oxytocincells in the dorsal part. The data demonstrated that the special distributed patterns of vasopressin- and oxytocin-neuron network have formed in those two nuclei at least from preterm. Intracerebroventricular injection of angiotensin II significantly increased fetal plasma oxytocin and vasopressin levels at preterm, which was associated with an increase of oxytocin- and vasopressin-neuron activity marked with c-fos expression. The data provided new evidence for the structural and functional development of the oxytocin- and vasopressin-network before birth., L. Shi, Y. Fan, Z. Xu., and Obsahuje seznam literatury
b1_We newly elaborated and adapte d several radiometric enzyme assays for the determination of activities of the key enzymes engaged in the biosynthesis (thyroid peroxidase, TPO) and metabolic transformations (conjugating enzymes and iodothyronine deiodinases, IDs) of thyroid hormones (THs) in the thyroid gland and in peripheral tissues, especially in white adipose tissue (WAT). We also elaborated novel, reliable radiometric methods for extremel y sensitive determination of enzyme activities of IDs of types 1, 2 and 3 in microsomal fractions of different rat and hum an tissues, as well as in homogenates of cultured mammalia n cells. The use of optimized TLC separation of radioactive products from the unconsumed substrates and film-less autoradiography of radiochromatograms, taking advantage of storage phosphor screens, enabled us to determine IDs enzyme activities as low as 10-18 katals. In studies of the interaction of fluoxetine (Fluox) with the metabolism of THs, we applied adapted radiometric enzyme assays for iodothyronine sulfotransferases (ST) and uridine 5’-diphosphoglucuronyltransferase (UDP-GT). Fluox is the most frequently used representative of a new group of non-tricyclic antidepressant drugs - selective serotonin re-uptake inhibitors. We used the elaborated assays fo r quantification the effects of Fluox and for the assessment of th e degree of potential induction of rat liver ST and/or UDP-GT enzyme activities by Fluox alone or in combination with T3 . Furthermore, we studied possible changes in IDs activities in murine adipose tissue under the conditions that promoted either tissue hypertrophy (obesogenic treatment) or involution (caloric restriction), and in response to leptin, using our newly developed radiometric enzyme assays for IDs., b2_Our results suggest that deiodinase D1 has a functional role in WAT, with D1 possibly being involved in the control of adipose tissue metabolism and/or accumulation of the tissue. Significant positive correlation between specific enzyme activity of D1 in WAT and plasma leptin levels was found. The newly developed and adapted radiometric enzyme assays proved to be very useful tools for studies of factors modulating THs metabolism, not only in model animals but also in clinical studies of human obesity., S. Pavelka., and Obsahuje bibliografii a bibliografické odkazy
Inhibitory neurotransmission plays a substantial role in encoding of auditory cues relevant for so und localization in vertebrates. While the anatomical organization of the respective afferent auditory brainstem circuits shows remarkable similarities between mammals and birds, the properties of inhibitory neurotransmission in these neural circuits are strikingly different. In mammals, inhibition is predom inantly glyciner gic and endowed with fast kinetics. In birds, inhibition is mediated by γ - Aminobutiric acid (GABA) and too slow to convey temporal information. A further prominent difference lies in the mechanism of inhibition in the respective systems. In auditory brainstem neurons of mammals, [Cl-] i undergoes a developmental shift causing the actions of GABA and glycine to gradually change from depolarization to the ‘classic’ hyperpolarizing-inhibition before hearing onset. Contrary to this, in the mature avian auditory brainstem Cl - homeostasis mechanisms accurately adjust the Cl - gradient to enable depolarizing, but still very efficient, shunting inhibition. The present review considers the mechanisms underlying development of the Cl - homeostasis in the auditory system of mammals and birds and discusses some open issues that require closer attention in future studies., I. Milenković, R. Rübsamen., and Obsahuje bibliografii a bibliografické odkazy
The aim of the study was to assess the impact of preweaning overnutrition upon the ontogeny of intestinal microbiota, alkaline phosphatase activity (AP) and pa rameters of growth and obesity in male Sprague-Dawley rats. We tested whether intestinal characteristics acquired in suckling pups could programme the development of enhanced fat deposition during normalized nutrition beyond weaning. Postnatal nutrition was manipulated by adjusting the number of pups in the nest to 4 (small litters - SL) and 10 (normal litters - NL). In the postweaning period both groups were fed with a standard diet. The jejunal and colonic Lactobacillus/Enterococcus (LAB) and the Bacteroides/Prevotella (BAC) were determined using the FISH technique, and the jejunal AP activity was assayed histochemically. At 15 and 20 days of age the SL pups became heavier, displayed increased adiposity accompanied by significantly higher LAB and lower numbers of BAC and with higher AP activity in comparison with rats nursed in NL nests. These differences pers isted to day 40 and withdrawal of the previous causal dietary influence did not prevent the post-weaning fat accretion. These results reveal the significance of early nutritional imprint upon the gut microbial/functional development and allow better unde rstanding of their involvement in the control of obesity., Z. Šefčíková ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Opening of the mitochondrial membrane permeability transition pore (MPTP) is an important factor in the activation of apoptotic and necrotic processes in mammalian cells. In a previous paper we have shown that cardiac mitochondria from neonatal rats are more resistant to calcium load than mitochondria from adult animals. In this study we have analyzed the ontogenetic development of this parameter both in heart and in liver mitochondria. We found that the high resistance of heart mitochondria decreases from day 14 to adulthood. On the other hand, we did not observe a similar age-dependent sensitivity in liver mitochondria, particularly in the neonatal period. Some significant but relatively smaller increase could be observed only after day 30. When compared with liver mitochondria cardiac mitochondria were more resistant also to the peroxide activating effect on calcium-induced mitochondrial swelling. These data thus indicate that the MPTP of heart mitochondria is better protected against damaging effects of the calcium load and oxidative stress. We can only speculate that the lower sensitivity to calcium-induced swelling may be related to the higher ischemic tolerance of the neonatal heart., Z. Drahota, ... [et al.]., and Obsahuje seznam literatury
The aim of our work was to provide data from women of fertile age with type 1 diabetes mellitus about the endogenous androgens and on their relations to the parameters of diabetes control. Forty-two women were examined, they did not use contraceptives for at least three months prior to the examination. A multivariate regression analysis showed that the daily insulin dose, the fasting glycemia and the HbA1c values and patient´s age correlated negatively with dehydroepiandrosterone sulfate, dehydroepiandrosterone and prolactin levels. The testosterone/ dehydroepiandrosterone sulfate ratio correlated positively with daily insulin dose and patient´s age. In contrast to adrenal androgens the values of other hormones, including total and free testosterone, androstenedione, dihydrotestosterone, estradiol, LH, FSH, 17-OH-P, progesterone and cortisol revealed no significant correlation. To conclude, significant relations between the glucose control parameters and the adrenal androgens and prolactin were demonstrated. These relationships should be considered as an important factor influencing diabetes control so the additional cardiovascular risk in women with DM1., H. Kvasnickova, R. Hampl, K. Vondra., and Obsahuje bibliografii
We have examined the changes of intercellular electrical coupling protein connexin-43 (Cx43) and of PKC-ε in heart atria of diabetic rats and/or after the treatment with triiodothyronine (T3 ). Diabetes was induced in Wistar-Kyoto rats by streptozotocin (50 mg/kg, i.v.) and atria were examined after 5 (acute stage) and 10 (chronic stage) weeks. T 3 (10 μg/100 g/day) was applied via a gastric tube for the last 10 days prior to the end of the experiments to non-diabetic and to the half of diabetic rats. Expression and phosphorylated status of Cx43, as well as expression of PKC-ε , were analyzed by Western blots using mouse monoclonal anti-Cx43 and rabbit polyclonal anti-PKC-ε antibodies. We found that the Cx43 expression was significantly increased after the treatment with T3 and in the acute diabetes. Both in diabetes and after T3 treatment the phosphorylation of Cx43 isoforms was markedly suppressed compared to the non-diabetic and T3-untreated controls. Such a down-regulation was less pronounced in diabetic rats after the T3-treatment. The expression of atrial PKC-ε was increased in diabetic rats. This increase was suppressed after T3 administration and the expression was decreased in T3-treated non-diabetic rats. We suggest that the reduced Cx43 phosphorylation in diabetic and hyperthyroid rats can deteriorate a cell-to-cell coupling and consequently facilitate a development of atrial tachyarrhythmia in diabetic or hyperthyroid animals., M. Mitašíková ... [et al.]., and Obsahuje seznam literatury
Decreased baroreflex sensitivity is an early sign of autonomic dysfunction in patients with type-1 diabetes mellitus. We evaluated the repeatability of a mild baroreflex sensitivity decrease in diabetics with respect to their heart rate. Finger blood pressure was continuously recorded in 14 young diabetics without clinical signs of autonomic dysfunction and in 14 age-matched controls for 42 min. The recordings were divided into 3-min segments, and the mean inter-beat interval (IBI), baroreflex sensitivity in ms/mm Hg (BRS) and mHz/mm Hg (BRSf) were determined in each segment. These values fluctuated in each subject within 42 min and therefore coefficients of repeatability were calculated for all subjects. Diabetics compared with controls had a decreased mean BRS (p=0.05), a tendency to a shortened IBI (p=0.08), and a decreased BRSf (p=0.17). IBI correlated with BRS in diabetics (p=0.03); th is correlation was at p=0.12 in the controls. BRSf was IBI independent (controls: p=0.81, diabetics: p=0.29). We conclude that BRS is partially dependent on mean IBI. Thus, BRS reflects not only an impairment of the quick baroreflex responses of IBI to blood pressure changes, but also a change of the tonic sy mpathetic and pa rasympathetic heart rate control. This is of significance during mild changes of BRS. Therefore, an examination of the BRSf index is highly recommended, because this examin ation improves the diagnostic value of the measurement, particul arly in cases of early signs of autonomic dysfunction., J. Svačinová ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy