1,721,006 research outputs found
Unique characteristics of Ca2+ homeostasis of the trans-Golgi compartment.
No full textTaking advantage of a fluorescent Ca(2+) indicator selectively targeted to the trans-Golgi lumen, we here demonstrate that its Ca(2+) homeostatic mechanisms are distinct from those of the other Golgi subcompartments: (i) Ca(2+) uptake depends exclusively on the activity of the secretory pathway Ca(2+) ATPase1 (SPCA1), whereas the sarco-endoplasmic reticulum Ca(2+) ATPase (SERCA) is excluded; (ii) IP(3) generated by receptor stimulation causes Ca(2+) uptake rather than release; (iii) Ca(2+) release can be triggered by activation of ryanodine receptors in cells endowed with robust expression of the latter channels (e.g., in neonatal cardiac myocyte). Finally, we show that, knocking down the SPCA1, and thus altering the trans-Golgi Ca(2+) content, specific functions associated with this subcompartment, such as sorting of proteins to the plasma membrane through the secretory pathway, and the structure of the entire Golgi apparatus are dramatically altered
INTRACELLULAR CA2+ STORES OF RAT CEREBELLUM - HETEROGENEITY WITHIN AND DISTINCTION FROM ENDOPLASMIC-RETICULUM
No full textRat cerebellum microsomes were subfractionated on isopycnic linear sucrose (20-42%)-density gradients. The distribution of endoplasmic reticulum (ER) markers (RNA, signal-sequence receptor alpha, calnexin, calreticulin, the immunoglobulin-binding protein Bip) and markers of intracellular rapidly exchanging Ca2+ stores [Ca2+ channels sensitive to either Ins(1,4,5)P3 or ryanodine) was investigated biochemically and immunologically. The comparison indicates that: (a) vesicles bearing the InsP3 receptor were separated from those bearing the ryanodine receptor; (b) ER markers, i.e. Bip, calnexin, signal-sequence receptor alpha, RNA, did not sediment as either InsP3 or ryanodine receptors did; (c) calreticulin, an intralumenal low-affinity high-capacity Ca(2+)-binding protein, had a widespread distribution, similar to that of Bip and calnexin, and was present in Purkinje, granule, Golgi and stellate neurons, as indicated by immunofluorescent labelling of cerebellum cortex cryosections. The present results show that the ER is not a homogeneous entity, and that Ca2+ stores are heterogeneous insofar as InsP3 receptors and ryanodine receptors are segregated, either to discrete intracellular organelles or to specialized ER subcompartment
A Vibrating Wire Susceptometer with a special electronic control for fast measurements at high temperatures
No full textThe measurement of the temperature behaviour of initial magnetic susceptibility is a powerful method for the thermomagnetic analysis of ferromagnetic materials. However, its application to nanostructured materials with technical relevance, particularly in the case of metastable systems, is made difficult by several conflicting conditions: the necessity to employ low magnetic fields, the required high sensitivity and the need for rapid scans in the high-temperature range. The vibrating wire susceptometer, an instrument belonging to the class of alternating gradient force magnetometers, has, in theory, the right characteristics to make such measurements. However, management of the instrument when carrying out rapid scans is intrinsically complex and requires a special electronic controller described here in detail. A combination of two phase-locked loop blocks is needed to provide the correct phase shift to ensure the locking of the resonance frequency while the instrument is working. A new measurement procedure that keeps the oscillation amplitude constant has also been implemented and it has proved to be very useful for rapid overview of the sample magnetic properties. The limitations of the controller performance due to the presence of noise are discussed. Extensive test measurements were carried out and analysed
Phase and frequency control in the vibrating wire magnetic susceptometer
No full textThe recently developed vibrating wire susceptometer (VWS) is an instrument inherently suitable to perform measurements with high sensitivity in a wide temperature range (up to 1200 K). In the present work we describe the principle of the electronic control of phase and frequency of the VWS that allows accurate determination of both the real and imaginary part of the susceptibility. In particular, the high quality factor, intrinsic to the mechanical system of the VWS, allows in principle to detect very small energy losses due to magnetic dissipative processes in the specimen
HETEROGENEITY OF MICROSOMAL CA2+ STORES IN CHICKEN PURKINJE NEURONS
No full textChicken cerebellum microsomes were subfractionated on isopycnic, linear sucrose (15-50%) density gradients. The distribution of four markers of intracellular, rapidly-exchanging Ca2+ stores, i.e. the Ca2+ pump, the receptors for inositol 1,4,5-trisphosphate (IP3) and ryanodine (Ry), and calsequestrin (CS, an intralumenal, high capacity Ca2+ binding protein) was investigated biochemically and immunologically. In the cerebellum, high levels of these markers are expressed by one of the cell types, the Purkinje neuron. Heavy subfractions were enriched in both CS and Ry receptor, intermediate subfractions in the IP3 receptor, while the Ca2+ pump was present in both intermediate and heavy subfractions. Intact cells and pelleted subfractions were examined by conventional and immuno-electron microscopy (immunogold labeling of ultrathin cryosections with anti-CS and anti-IP3 receptor antibodies). Of the strongly CS-labeled, moderately dense-cored vacuoles (calciosomes) recently described in chicken Purkinje neurons only partly exhibited labeling for the IP3 receptor as well, and the rest appeared negative. The latter were enriched in a heavy subfraction of the gradient where Ry receptors were also concentrated, whereas the CS-rich vacuoles in an intermediate subfraction were almost always IP3 receptor-positive. The population of CS-rich calciosomes of chicken Purkinje neurons appears therefore to be molecularly heterogeneous, with a part responsive to IP3 and the rest possibly sensitive to Ry
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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