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1.
Maria Weinert Tharakeswari Selvakumar Travis S. Tierney Kambiz N. Alavian 《Journal of visualized experiments : JoVE》2015,(96)
Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson’s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population. 相似文献
2.
《Peptides》2015
A new peptide with 61 amino acids cross-linked by 4 disulfide bridges, with molecular weight of 6938.12 Da, and an amidated C-terminal amino acid residue was purified and characterized. The primary structure was obtained by direct Edman degradation and sequencing its gene. The peptide is lethal to mammals and was shown to be similar (95% identity) to toxin Ts1 (gamma toxin) from the Brazilian scorpion Tityus serrulatus; it was named Tt1g (from T. trivittatus toxin 1 gamma-like). Tt1g was assayed on several sub-types of Na+-channels showing displacement of the currents to more negative voltages, being the hNav1.3 the most affected channel. This toxin displays characteristics typical to the β-type sodium scorpion toxins. Lethality tests and physiological assays indicate that this peptide is probably the most important toxic component of this species of scorpion, known for causing human fatalities in the South American continent. 相似文献
3.
Summary Marion Island (47°S, 38°E) has one of the most oceanic climates on earth, with consistently low air temperatures, high precipitation, constantly high humidity, and low incident radiation. Since 1968 mean surface air temperature has increased significantly, by 0.025° C year–1. This was strongly associated with corresponding changes in sea surface temperature but only weakly, or not at all, with variations in radiation and precipitation. We suggest that changing sealevel (atmospheric and oceanic) circulation patterns in the region underlie all of these changes. Sub-Antarctic terrestrial ecosystems are characterized by being species-poor and having a simple trophic structure. Marion Island is no exception and a scenario is presented of the implications of climatic change for the structure and functioning of its ecosystem. Primary production on the island is high and consequently the vegetation has a large annual requirement for nutrients. There are no macroherbivores and even the insects play only a small role as herbivores, so most of the energy and nutrients incorporated in primary production go through a detritus, rather than grazing, cycle. Ameliorating temperatures and increasing CO2 levels are expected to increase productivity and nutrient demand even further. However, most of the plant communities occur on soils which have especially low available levels of nutrients and nutrient mineralization from organic reserves is the main bottleneck in nutrient cycling and primary production. Increasing temperatures will not significantly enhance microbially-mediated mineralization rates since soil microbiological processes on the island are strongly limited by waterlogging, rather than by temperature. The island supports large numbers of soil macro-arthropods, which are responsible for most of the nutrient release from peat and litter. The activities of these animals are strongly temperature dependent and increasing temperature will result in enhanced nutrient availability, allowing the potential for increased primary production due to elevated temperature and CO2 levels to be realized. However, housemice occur on the island and have an important influence on the ecosystem, mainly by feeding on soil invertebrates. The mouse population is strongly temperature-limited and appears to be increasing, possibly as a result of ameliorating temperatures. We suggest that an increasing mouse population, through enhanced predation pressure on soil invertebrates, will decrease overall rates of nutrient cycling and cause imbalances between primary production and decomposition. This, along with more direct effects of mice (e.g. granivory) has important implications for vegetation succession and ecosystem structure and functioning on the island. Some of these are already apparent from comparisons with nearby Prince Edward Island where mice do not occur. Other implications of climatic change for the island are presented which emphasize the very marked influences that invasive organisms have on ecosystem structure and functioning. We suggest that changing sealevel circulation patterns, by allowing opportunities for colonization by new biota, may have an even more important influence on terrestrial sub-Antarctic ecosystems than is suggested merely on the basis of associated changes in temperature or precipitation. 相似文献
4.
Fire can cause severe nitrogen (N) losses from grassland, chaparral, and temperate and boreal forest ecosystems. Paradoxically,
soil ammonium levels are markedly increased by fire, resulting in high rates of primary production in re-establishing plant
communities. In a manipulative experiment, we examined the influence of wild-fire ash residues on soil, microbial and plant
N pools in a recently burned Californian bishop pine (Pinus muricata D. Don) forest. Ash stimulated post-fire primary production and ecosystem N retention through direct N inputs from ash to
soils, as well as indirect ash effects on soil N availability to plants. These results suggest that redistribution of surface
ash after fire by wind or water may cause substantial heterogeneity in soil N availability to plants, and could be an important
mechanism contributing to vegetation patchiness in fire-prone ecosystems. In addition, we investigated the impact of fire
on ecosystem N cycling by comparing 15N natural abundance values from recently burned and nearby unburned P. muricata forest communities. At the burned site, 15N natural abundance in recolonising species was similar to that in bulk soil organic matter. By contrast, there was a marked
15N depletion in the same species relative to the total soil N pool at the unburned site. These results suggest that plant uptake
of nitrate (which tends to be strongly depleted in 15N because of fractionation during nitrification) is low in recently burned forest communities but could be an important component
of eco- system N cycling in mature conifer stands.
Received: 29 June 1999 / Accepted: 24 October 1999 相似文献
5.
B. Leroi 《Population Ecology》1973,15(1):163-182
The leaf-mining larvae of the celery fly, P. heraclei, have the ability to leave their primary mine to bore a secondary mine in another leaflet or leaf. This phenomenon is always numerically significant under natural conditions (depending on the plots and the years, 38 to 97% of the larvae observed made such a change), although it is not an obligatory behavior. The distribution of the secondary mines on the leaflets and on the leaves varies with the importance of the migrations, with the total unoccupied and healthy leaf area and with the relative position of different leaves on the plant. These migrations are due to the insufficiency of healthy food available for the larvae: either because the quantity of parenchyma offered by a leaflet is too small for a larva, or because there is intraspecific competition following multiple egg-layings on the same leaflet, or because there is a deterioration of the quality of the parenchyma, particularly following the development of celery leaf-spot. The possibility for the larvae of P. heraclei to bore secondary mines appears to be an extremely important factor in the population dynamics of this species. This permits the reduction of the negative effects of intraspecific competition and celery leaf-spot, and permits the colonization by the larvae of young leaves not used by the females at the time of egg-laying. 相似文献
6.
Summary . We propose a fully inferential model-based approach to the problem of comparing the firing patterns of a neuron recorded under two distinct experimental conditions. The methodology is based on nonhomogeneous Poisson process models for the firing times of each condition with flexible nonparametric mixture prior models for the corresponding intensity functions. We demonstrate posterior inferences from a global analysis, which may be used to compare the two conditions over the entire experimental time window, as well as from a pointwise analysis at selected time points to detect local deviations of firing patterns from one condition to another. We apply our method on two neurons recorded from the primary motor cortex area of a monkey's brain while performing a sequence of reaching tasks. 相似文献
7.
The metallochromic indicator murexide has been used to monitor calcium concentration changes during the dextran-induced, phosphatidylserine-dependent degranulation of rat peritoneal mast cells. The dextran-induced Ca2+-uptake showed an absolute dependence on the presence of phosphatidylserine. The extent of Ca2+-uptake increased with phosphatidylserine in a concentration-dependent manner. At 25 degrees C the half-life of the uptake process equalled 35 +/- 5 s. Exposure of the mast cells to dextran in the presence of Ca2+, but in the absence of phosphatidylserine, desensitized the cells. The subsequent addition of phosphatidylserine failed to restore the Ca2+-uptake activity. However, the Ca2+-ionophore A23187 did promote Ca2+ uptake by the cells without PS. 相似文献
8.
Maria Teresa de los Frailes Lucinda Cacicedo Maria Jesús Lorenzo Franco Sánchez-Franco 《Journal of neurochemistry》1989,52(5):1333-1339
The influence of membrane depolarization on somatostatin secretion and protein synthesis by fetal and neonatal cerebrocortical neurons was studied. Cortical cells obtained by mechanical dispersion were maintained as monolayer cultures for 8 days. The ability of fetal cerebrocortical and hypothalamic cells to release immunoreactive somatostatin (IR-SRIF) was confirmed. Total protein synthesis was determined by the incorporation of [3H]phenylalanine into trichloroacetic acid-precipitable proteins. To study the effect of acute depolarization on protein synthesis, cells were incubated for 30 min with [3H]phenylalanine or [3H]leucine and the depolarizing agent. In fetal cerebrocortical cells, potassium (30 and 56 mM) decreased protein synthesis and RNA levels and increased IR-SRIF release. Depolarization by veratridine, a sodium channel activator, induced a similar effect. The effect of veratridine on IR-SRIF and protein synthesis was reversed by tetrodotoxin, a sodium channel blocker, or verapamil, a calcium channel blocker. These findings suggest that protein synthesis by cerebrocortical cells is decreased in fetal brain cells by membrane depolarization and is dependent on Na+ and Ca2+ entry into cells. In postnatal (day 7) cerebrocortical cells, depolarization induced by high potassium concentrations led to a concomitant increase in protein synthesis, RNA content, and somatostatin release. These findings indicate that depolarization of the cellular membrane is coupled to an increase in protein synthesis in neonatal, but not in fetal, dispersed brain cells. 相似文献
9.
Aditya Mittal Anandkumar Madhavjibhai Changani Sakshi Taparia 《Journal of biomolecular structure & dynamics》2020,38(15):4579-4583
AbstractNumber of naturally occurring primary sequences of proteins is an infinitesimally small subset of the possible number of primary sequences that can be synthesized using 20 amino acids. Prevailing views ascribe this to slow and incremental mutational/selection evolutionary mechanisms. However, considering the large number of avenues available in form of diversity of emerging/evolving and/or disappearing living systems for exploring the primary sequence space over the evolutionary time scale of ~3.5 billion years, this remains a conjecture. Therefore, to investigate primary sequence space limitations, we carried out a systematic study for finding primary sequences absent in nature. We report the discovery of the smallest peptide sequence “Cysteine-Glutamine-Tryptophan-Tryptophan” that is not found in over half-a-million curated protein sequences in the Uniprot (Swiss-Prot) database. Additionally, we report a library of 83605 pentapeptides that are not found in any of the known protein sequences. Compositional analyses of these absent primary sequences yield a remarkably strong power relationship between the percentage occurrence of individual amino acids in all known protein sequences and their respective frequency of occurrence in the absent peptides, regardless of their specific position in the sequences. If random evolutionary mechanisms were responsible for limitations to the primary sequence space, then one would not expect any relationship between compositions of available and absent primary sequences. Thus, we conclusively show that stoichiometric constraints on amino acids limit the primary sequence space of proteins in nature. We discuss the possibly profound implications of our findings in both evolutionary and synthetic biology.Communicated by Ramaswamy H. Sarma 相似文献
10.
The effects of implementing Directive 91/271/EEC of 21 May 1991 (Waste Water Treatment Plan Directive) and Directive 91/676/EEC
of 12 December (Nitrates Directive) are analysed in 7 Portuguese estuaries (Minho, Lima, Douro, Mondego, Tagus, Sado and Guadiana)
and two coastal lagoons (Ria de Aveiro and Ria Formosa), with a modelling approach. MOHID Water Modelling System was used
to perform simulations with three nitrogen load scenarios for each system: a reference scenario, a 50% nitrate removal by
agriculture scenario and another with a 100% nutrients removal by waste water treatment plants (WWTP). It is shown that the
interaction between hydrodynamic and ecological processes is an important feature to study trophic problems in estuaries.
Ecological processes such as primary production only occur inside the system if the residence time of water is high enough
to enable organismal activity and if the adequate conditions are found (e.g. light, nutrients, temperature). From the model
results it is possible to conclude: (i) in systems with short residence time a reduction in nutrient load will only produce
a decrease in nutrient transit and will not affect the system’s global ecological status (e.g. Douro Estuary); (ii) in systems
with long residence time the effects will range from significant, when primary production is mostly limited by nutrients (e.g.
Ria de Aveiro), to non-significant, when primary production in the system is light-limited (e.g. Tagus Estuary). 相似文献