Larval Manduca prothoracic gland cells in vitro responded to prothoracicotropic hormone (PTTH) from neurosecretory cells of the brain with an increase of intracellular free calcium. This effect is reversible and dose-dependent. Preincubation of the glands with TMB-8 and dantrolene, which inhibit the release of calcium from intracellular stores, did not decrease the PTTH-stimulated increase in calcium, indicating that intracellular calcium stores are not involved in the control of ecdysteroidogenesis. Pharmacological studies of the PTTH effect with calcium channel blockers revealed that the increase in calcium was totally blocked by cadmium, partially inhibited by nickel and lanthanum and by amiloride, an antagonist of T-type calcium channels. All other inhibitors tested were ineffective, suggesting that the increase in cytosolic calcium is induced by opening of calcium channels, presumably of the T-type, in response to PTTH. The action of PTTH on these channels may be mediated by a G-protein as shown by the effect of mastoparan, a G-protein activator, which increased the concentration of cytosolic calcium comparable to that evoked by PTTH., Heiner Birkenbeil, and Lit
The nature and rate of juvenile hormone (JH) biosynthesis and effects of allatostatins and allatotropin have been investigated in isolated corpora allata (CA) of adults and larvae of the noctuid tomato moth, Lacanobia oleracea. In adult female CA, mean rates of synthesis were relatively constant (10-16 pmol/pr/h) at all times. However, the range of JH synthesis by individual CA of similarly aged insects was quite large (2-30 pmol/pr/h). High performance liquid chromatography (HPLC) separation shows that adult female moth CA synthesise predominantly JH I and JH II. Rates of JH synthesis in vitro are dependent on methionine concentration. Synthetic Manduca sexta allatostatin (Mas-AS) caused a dose-dependent inhibition of JH synthesis by adult female CA but only to a max. of 54%, whilst 10 µM synthetic M. sexta allatotropin caused a 37% stimulation of CA activity. At 1 mM the cockroach allatostatin, Dip-allatostatin-2, had no significant effect on JH synthesis. In larval L. oleracea, rates of JH biosynthesis were very low., Neil Audsley, Robert J. Weaver, John P. Edwards, and Lit
The Laboratory of Cell Regeneration and Plasticity and the PIGMOD Centre of the Institute of Animal Physiology and Genetics of the CAS in Liběchov have been using their unique biomedical models - miniature pigs - to study fundamental causes of serious diseases, identify new biomarkers and search for new therapeutic methods /including those based on cell therapy and genetic therapy) to treat amyotrophic lateral sclerosis, traumatic spinal cord injury and human melanoma. Special transgenic minipigs carrying human mutated huntingtin allow researchers to investigate in detail the fatal neurodegenerative disorder called Huntington's disease. and Jana Olivová.
Climate features that influence life cycles, notably severity, seasonality, unpredictability and variability, are summarized for different polar zones. The zones differ widely in these factors and how they are combined. For example, seasonality is markedly reduced by oceanic influences in the Subantarctic. Information about the life cycles of Arctic and Antarctic arthropods is reviewed to assess the relative contributions of flexibility and programming to life cycles in polar regions. A wide range of life cycles occurs in polar arthropods and, when whole life cycles are considered, fixed or programmed elements are well represented, in contrast to some recent opinions that emphasize the prevalence of flexible or opportunistic responses. Programmed responses ale especially common for controlling the appearance of stages that are sensitive to adverse conditions, such as the reproductive adult. The relative contribution of flexibility and programming to different life cycles is correlated with taxonomic affinity (which establishes the general lifecycle framework for a species), and with climatic zone, the habitats of immature and adult stages, and food., Hugh V. Danks, and Lit
We have separated 2b myosin heavy chain (MyHC) isoform from the rat extensor digitorum longus muscle by SDS-PAGE and analyzed it by two subsequent mass spectrometry techniques. After tryptic digestion, the obtained peptides were identified by Matrix-Assisted Laser Desorption/Ionisation reflectron Time of Flight mass spectrometry (MALDI-TOF MS) and sequenced by Liquid chromatography tandem mass spectrometry (ESI LC/MS/MS). The analyzed peptides proportionally covered 30 % of the 2b MyHC isoform sequence. The results suggest that the primary structure is identical with the highest probability to a NCBI database record ref|NP_062198.1|, representing the last updated record of rat 2b isoform. Nonetheless, four peptides carrying amino acid substitution(s) in comparison with the NCBI database record were identified., J. Žurmanová, D. Maláčová, F. Půta, P. Novák, J. Říčný, T. Soukup., and Obsahuje bibliografii a bibliografické odkazy
Glutamate is a well-characterized excitatory neurotransmitter in the central nervous system (CNS). Recently, glutamate receptors (GluRs) were also found in peripheral tissues, including the heart. However, the function of GluRs in peripheral organs remains poorly understood. In the present study, we found that N-methyl-D-aspartate (NMDA) could increase intracellular calcium ([Ca2+]i) level in a dose-dependent manner in cultured neonatal rat cardiomyocytes. NMDA at 10-4 M increased the levels of reactive oxygen species (ROS), cytosolic cytochrome c (cyto c), and 17-kDa caspase-3, but depolarized mitochondrial membrane potential, leading to cardiomyocyte apoptosis. In addition, NMDA treatment induced an increase in ba x mRNA but a decrease in bcl-2 mRNA expression in the cardiomyocytes. The above effects of NMDA were bloc ked by the NMDA receptor antagonist (+)-5-methyl- 10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801), and by ROS scavengers glutathione (GSH) and N-acetylcystein (NAC). These results suggest that stimulation of NMDA receptor in the cardiomyocyte may lead to apoptosis via a Ca2+, ROS, and caspase-3 mediated pathway. These findings suggest that NMDA receptor may play an important role in myocardial pathogenesis., X. Gao, X. Xu, J. Pang, C. Zhang, J. M. Ding, X. Peng, Y. Liu, J.-M. Cao., and Obsahuje bibliografii a bibliografické odkazy
The aim of this work was to evaluate ontogeny of reactive nitrogen species (RNS) production by peripheral blood phagocytes in pig. Pig fetuses (55 and 92 days of gestation) and postnatal piglets (1, 3, 8, 17, 31 and 41 days after birth) were used. RNS production was measured by fluorescent probes diaminofluorescein-diacetate (DAF-FMDA) and dichloro-fluorescein-diacetate (H2DCFDA). Levels of nitration of cell proteins were established by immunofluorescent detection of nitrotyrosine. Levels of plasma nitrites/nitrates were detected spectrophotometrically by Griess reaction. Nitric oxide production measured by DAF-FMDA in neutrophils decreased during postnatal life. Spontaneous RNS measured by H2DCFDA decreased from 55th day of gestation to the 41st day of life. Phorbol-12-myristate-13-acetate activated production decreased during postnatal life. Production of NO measured by DAF-FMDA in macrophages decreased from the 1st to 41st day after birth. RNS production measured by H2DCFDA in monocytes did not show any significant changes during ontogeny. The level of nitrotyrosine significantly decreased from the 3rd to 17th day. Levels of plasma nitrites/nitrates gradually decreased from the 55th day of gestation to the 41st day after birth. A temporary increase in all parameters occurred after weaning, but without any significance. In conclusion, RNS production has a decreasing trend during ontogeny and is transiently upregulated after weaning., P. Zelníčková, M. Faldyna, J. Ondráček, H. Kovářů, F. Kovářů., and Obsahuje bibliografii a bibliografické odkazy
First stadium juveniles of P. angustus were reared under controlled seasonal conditions to maturity, reproduction and death. Individuals born in any one breeding season either had a 1-year or a 2-year life cycle (cohort-splitting). The life cycle was annual for individuals born in the first part of the breeding season (May-August), but became biennial for those born later (August-October). Two phenomena were involved: (1) Only individuals reaching the penultimate stadium (stadium VII) before a critical period at the end of spring could become adult in the breeding season following that of their birth. After this time, stadium VII individuals entered into aestivation and only became adult in the second autumn after their birth. (2) Females becoming adult in autumn entered reproductive dormancy and only laid eggs in the following spring. Overall, individuals born at the start of the breeding season easily reached stadium VII before the critical period and were able to breed at I year, whereas individuals born at the end of the breeding season reached stadium VII after the critical period, then had two consecutive periods of dormancy and only bred at 2 years age. Individuals from the same nest born in the middle of the breeding season (August) could have either annual or biennial life cycles, depending on whether they reached stadium VII before or during aestivation. The environmental factors capable of triggering aestivation in subadults and reproductive dormancy in autumn-maturing females are discussed., Jean-Francois David, Marie-Louise Celerier, Jean-Jacques Geoffroy, and Lit
To understand the contribution of potassium (K+) channels, particularly α-dendrotoxin (D-type)-sensitive K+ channels (Kv.1, Kv1.2 or Kv1.6 subunits), to the generation of neuronal spike output we must have detailed information of the functional role of these channels in the neuronal membrane. Conventional intracellular recording methods in current clamp mode were used to identify the role of α-dendrotoxin (α-DTX)-sensitive K+ channel currents in shaping the spike output and modulation of neuronal properties of cerebellar Purkinje neurons (PCs) in slices. Addition of α-DTX revealed that D-type K+ channels play an important role in the shaping of Purkinje neuronal firing behavior. Repetitive firing capability of PCs was increased following exposure to artificial cerebrospinal fluid (aCSF) containing α-DTX, so that in response to the injection of 0.6 nA depolarizing current pulse of 600 ms, the number of action potentials insignificantly increased from 15 in the presence of 4-AP to 29 action potentials per second after application of DTX following pretreatment with 4-AP. These results indicate that D-type K+ channels (Kv.1, Kv1.2 or Kv1.6 subunits) may contribute to the spike frequency adaptation in PCs. Our findings suggest that the activation of voltage-dependent K+ channels (D and A types) markedly affect the firing pattern of PCs., H. Haghdoust, M. Janahmadi, G. Behzadi., and Obsahuje bibliografii a bibliografické odkazy