Merging of <Emphasis Type="Italic">Pedicularis exaltata</Emphasis> and <Emphasis Type="Italic">P. hacquetii</Emphasis> in the Carpathians: from local history to regional phylogeography based on complex evidence

The disjunct occurrence of Pedicularis exaltata in the White Carpathians (Czech Republic), isolated by more than 500 km from the nearest populations in the Eastern Carpathians, has been considered one of the mysteries of the Western Carpathian flora. We used molecular methods (AFLP, ITS and cpDNA sequencing) to reconstruct a possible scenario of the evolution of P. exaltata and its closely related congener P. hacquetii and to evaluate their differentiation. We paid particular attention to the origin of the isolated population in the White Carpathians. We also analysed the vegetation composition at the sampling sites to characterize the habitat preferences of the Pedicularis species and compare different sites. The pattern of molecular variability does not support a species boundary between P. exaltata and P. hacquetii. These assumed species should be merged into one bearing the name Pedicularis hacquetii following the priority rule, as also proposed by several early morphology-based taxonomic studies. The cpDNA variability pattern supports an evolutionary scenario involving a distribution centre (refugium) in the Eastern Carpathians from which the species expanded to the more westerly parts of its extant geographical range. Low population differentiation in AFLPs, high gene diversity and high DW index in isolated populations indicate that this expansion was contiguous rather than based on long-distance dispersal. Under such a scenario, the White Carpathian population is considered one of isolated relict populations scattered throughout the Carpathians and the East European Plain. A complete phylogeographic reconstruction of P. hacquetii s.l., however, requires ancient DNA analysis of herbarium specimens of nowadays extinct populations of the East European Plain. Extant habitats of P. hacquetii s.l. consist of species-rich dry-mesic to mesic (sub)montane grasslands and subalpine tall-herb growths, which may be relics of vegetation widespread in the late Glacial/early Holocene, when the contiguous expansion of the species possibly proceeded. The analysis of species with high fidelity to P. hacquetii s.l. throughout its range indicated that middle altitudes of south-western White Carpathians belong to the regions most suitable for the taxon within the Czech Republic and Slovakia. We found no support for the late introduction/long-distance dispersal scenarios and consider the relic survival scenario most parsimonious for the White Carpathians.     

Skin aging: Dermal adipocytes metabolically reprogram dermal fibroblasts

Emerging data connects the aging process in dermal fibroblasts with metabolic reprogramming, provided by enhanced fatty acid oxidation and reduced glycolysis. This switch may be caused by a significant expansion of the dermal white adipose tissue (dWAT) layer in aged, hair-covered skin. Dermal adipocytes cycle through de-differentiation and re-differentiation. As a result, there is a strongly enhanced release of free fatty acids into the extracellular space during the de-differentiation of dermal adipocytes in the catagen phase of the hair follicle cycle. Both caveolin-1 and adiponectin are critical factors influencing these processes. Controlling the expression levels of these two factors also offers the ability to manipulate the metabolic preferences of the different cell types within the microenvironment of the skin, including dermal fibroblasts. Differential expression of adiponectin and caveolin-1 in the various cell types may also be responsible for the cellular metabolic heterogeneity within the cells of the skin.     

Participation of Intracellular Ca2+ Stores in Ca2+ Signalling in Neurons of the Thalamic Laterodorsal Nucleus of Rats

Experiments were carried out on isolated neurons of the thalamic nucleus lateralis dorsalis (LD) from 12-day-old rats. According to the morphological characteristics, LD neurons were classified as relay thalamo-cortical units and interneurons. The concentration of free Ca²⁺ ions in the cytoplasm ([Ca²⁺] _i ) was measured by a fluorescent calcium indicator, fura-2AM. Application of 30 mM caffeine caused a transient change in the [Ca²⁺] _i in 8 of 15 and in 6 of 11 of the thalamo-cortical units and interneurons under study, respectively. After stimulation of a cell with application of 50 mM KCl, a caffeine-induced increase in the [Ca²⁺] _i was observed in all tested neurons. To study the contribution of Ca²⁺-induced Ca²⁺ release (CICR) to the calcium transient evoked by depolarization of the neuronal membrane, caffeine in a subthreshold concentration was pre-applied. After 50 mM KCl had been added to the medium following pre-application of 0.5 mM caffeine, the calcium transient amplitude in thalamo-cortical neurons increased by 51 ± 7% (n = 16). In interneurons this effect was not observed (n = 11). The data obtained allow us to hypothesize that CICR contributes to the depolarization-evoked calcium transient only in the relay (thalamo-cortical) neurons. Differences in the pattern of calcium signalling, which were detected in two types of neurons of the thalamic LD, can be a factor determining distinctions in the physiological characteristics of these neurons.     

The noradrenaline and serotonin content in symmetrical parts of the normal rat brain and during learning and peptide administration]

Content was compared of noradrenaline (NA) and serotonine (5-OT) in the right and left halves of the rats brain in norm, at elaboration of defensive conditioned reflexes of two-ways avoidance (CRTWA) and at administration of neuropeptides influencing the learning and memory--dezglycilargininvasopressin (DG-AVP), ACTH4-7 pro-gli-pro and dalargin. The conducted studies showed that in control animals the content of NA in the cortex of the right hemisphere was significantly higher than in the cortex of the left one. For the content of 5-OT in symmetric brain parts no significant differences were revealed. Under the elaboration of CRTWA the asymmetry of NA content was not eliminated. Systemic administration of DG-AVP, ACTG4-7 pro-gli-pro and dalargin practically did not change the content of 5-OT, but reduced the content of NA in the cortex and the rest of the brain, and the content of NA in the right and left cortex was equalized. The obtained data point to the asymmetric character of neuropeptides action and to greater resistance of 5-OT-ergic brain system to functional load and to administration of peptides in comparison with NA-ergic system.     

Relationship between noradrenalin uptake and secretion by cortical nerve endings in rats during conditioning

The intensity of highly selective uptake (inactivation) of noradrenalin (NA) and of spontaneous and evoked secretion by synaptosomes in the rat cerebral cortex were investigated by the use of [³H]-NA 30 min after defensive conditioning. Changes in the efficiency of adrenergic synaptic transmission during formation and fixation of the temporary connection were judged on the basis of these parameters. Changes in uptake and spontaneous secretion were not observed in the trained and control rats, whereas the intensity of its secretion induced by KC1 (60 mM) was considerably higher in the trained rats than in the controls. An increase in the intensity of evoked secretion was observed only during conditioned reflex formation. It is concluded that conditioning leads to a specific increase in the efficiency of adrenergic synaptic transmission, which is observable in nerve ending isolated 30 min after formation of the reflex.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR. Translated from Neirofiziologiya, Vol. 14, No. 4, pp. 367–372, July–August, 1982.     

Accounting for recovery processes in the description of interphase cell death

A phenomenological scheme is proposed to describe dose- and time-dependences of the interphase cell death in terms of a continual model of radiation injury. Different regularities of the postirradiation recovery of cells are discussed. The results obtained are in agreement with the experimental data.     

Changes in functioning of rat submandibular salivary gland under streptozotocin-induced diabetes are associated with alterations of Ca2+ signaling and Ca2+ transporting pumps

Xerostomia and pathological thirst are troublesome complications of diabetes mellitus associated with impaired functioning of salivary glands; however, their cellular mechanisms are not yet determined. Isolated acinar cells were loaded with Ca2+ indicators fura-2/AM for measuring cytosolic Ca2+ concentration ([Ca2+]i) or mag-fura-2/AM-inside the endoplasmic reticulum (ER). We found a dramatic decrease in pilocarpine-stimulated saliva flow, protein content and amylase activity in rats after 6 weeks of diabetes vs. healthy animals. This was accompanied with rise in resting [Ca2+]i and increased potency of acetylcholine (ACh) and carbachol (CCh) but not norepinephrine (NE) to induce [Ca2+]i transients in acinar cells from diabetic animals. However, [Ca2+]i transients mediated by Ca2+ release from ER stores (induced by application of either ACh, CCh, NE, or ionomycin in Ca2+-free extracellular medium) were decreased under diabetes. Application of inositol-1,4,5-trisphosphate led to smaller Ca2+ release from ER under the diabetes. Both plasmalemma and ER Ca2+-ATPases activity was reduced and the latter showed the increased affinity to ATP under the diabetes. We conclude that the diabetes caused impairment of salivary cells functions that, on the cellular level, associates with Ca2+ overload, increased Ca2+-mobilizing ability of muscarinic but not adrenergic receptors, decreased Ca2+-ATPases activity and ER Ca2+ content.     

Caffeine-induced calcium release from the endoplasmic reticulum of acinar cells of the submandibular salivary gland

Ryanodine receptors (RyRs) play a key role in the generalization and spreading of calcium waves in excitable cells; however, the question of the existence of functionally active RyRs in nonexcitable cells demonstrating the capacity for exocytosis (e.g., salivary gland acini) remains open. We studied changes in the total amount of calcium stored in the endoplasmic reticulum (ER) of acinar cells of the submandibular salivary gland of rats and changes in the concentration of ionized Ca²⁺ inside the ER ([Ca²⁺]_ER) using, respectively, a metallochrome dye, arsenazo III, and a low-affinity fluorescent dye, mag-fura 2/AM. In permeabilized cells, caffeine caused dose-dependent decreases in the total amount of calcium and concentration of ionized calcium. The effective concentration of caffeine providing a 50% drop in the [Ca²⁺]_ER (EC₅₀) was, on average, 7.3 ± 1.1 mM. The caffeine-induced drop in the [Ca²⁺]^ER was insensitive to heparin; in addition, it was blocked by high concentrations (100 μM) of ryanodine, potentiated by ryanodine applied in mild concentrations (10 μM), and also demonstrated a bell-shaped dependence on the concentration of cytoplasmic Ca²⁺. Such peculiarities are typical characteristics of the RyR-mediated reaction. Therefore, functional RyRs whose activation results in a transient release of calcium from the ER are present in acinar cells of the submandibular salivary gland. Neirofiziologiya/Neurophysiology, Vol. 39, No. 2, pp. 107–112, March–April, 2007.     

Mitochondria adjust Ca(2+) signaling regime to a pattern of stimulation in salivary acinar cells

The salivary acinar cells have unique Ca(2+) signaling machinery that ensures an extensive secretion. The agonist-induced secretion is governed by Ca(2+) signals originated from the endoplasmic reticulum (ER) followed by a store-operated Ca(2+) entry (SOCE). During tasting and chewing food a frequency of parasympathetic stimulation increases up to ten fold, entailing cells to adapt its Ca(2+) machinery to promote ER refilling and ensure sustained SOCE by yet unknown mechanism. By employing a combination of fluorescent Ca(2+) imaging in the cytoplasm and inside cellular organelles (ER and mitochondria) we described the role of mitochondria in adjustment of Ca(2+) signaling regime and ER refilling according to a pattern of agonist stimulation. Under the sustained stimulation, SOCE is increased proportionally to the degree of ER depletion. Cell adapts its Ca(2+) handling system directing more Ca(2+) into mitochondria via microdomains of high [Ca(2+)] providing positive feedback on SOCE while intra-mitochondrial tunneling provides adequate ER refilling. In the absence of an agonist, the bulk of ER refilling occurs through Ca(2+)-ATPase-mediated Ca(2+) uptake within subplasmalemmal space. In conclusion, mitochondria play a key role in the maintenance of sustained SOCE and adequate ER refilling by regulating Ca(2+) fluxes within the cell that may represent an intrinsic adaptation mechanism to ensure a long-lasting secretion.