Ca2+ signals mediated by Ins(1,4,5)P3-gated channels in rat ureteric myocytes

2000 ◽  
Vol 349 (1) ◽  
pp. 323-332 ◽  
Author(s):  
François-Xavier BOITTIN ◽  
Frédéric COUSSIN ◽  
Jean-Luc MOREL ◽  
Guillaume HALET ◽  
Nathalie MACREZ ◽  
...  
Keyword(s):  
Ryanodine Receptor ◽  
Time Course ◽  
Binding Capacity ◽  
Ryanodine Receptors ◽  
Initiation Site ◽  
Variable Number ◽  
Spatial Spread ◽  
Receptor Antibody ◽  
Low Concentrations ◽  
High Level

Localized Ca2+-release signals (puffs) and propagated Ca2+ waves were characterized in rat ureteric myocytes by confocal microscopy. Ca2+ puffs were evoked by photorelease of low concentrations of Ins(1,4,5)P3 from a caged precursor and by low concentrations of acetylcholine; they were also observed spontaneously in Ca2+-overloaded myocytes. Ca2+ puffs showed some variability in amplitude, time course and spatial spread, suggesting that Ins(1,4,5)P3-gated channels exist in clusters containing variable numbers of channels and that within these clusters a variable number of channels can be recruited. Immunodetection of Ins(1,4,5)P3 receptors revealed the existence of several spots of fluorescence in the confocal cell sections, supporting the existence of clusters of Ins(1,4,5)P3 receptors. Strong Ins(1,4,5)P3 photorelease and high concentrations of acetylcholine induced Ca2+ waves that originated from an initiation site and propagated in the whole cell by spatial recruitment of neighbouring Ca2+-release sites. Both Ca2+ puffs and Ca2+ waves were blocked selectively by intracellular applications of heparin and an anti-Ins(1,4,5)P3-receptor antibody, but were unaffected by ryanodine and intracellular application of an anti-ryanodine receptor antibody. mRNAs encoding for the three subtypes of Ins(1,4,5)P3 receptor and subtype 3 of ryanodine receptor were detected in these myocytes, and the maximal binding capacity of [3H]Ins(1,4,5)P3 was 10- to 12-fold higher than that of [3H]ryanodine. These results suggest that Ins(1,4,5)P3-gated channels mediate a continuum of Ca2+ signalling in smooth-muscle cells expressing a high level of Ins(1,4,5)P3 receptors and no subtypes 1 and 2 of ryanodine receptors.

Adaptation of the skeletal muscle calcium-release mechanism to weight-bearing condition

1996 ◽  
Vol 270 (6) ◽  
pp. C1588-C1594 ◽  
Author(s):  
S. C. Kandarian ◽  
D. G. Peters ◽  
T. G. Favero ◽  
C. W. Ward ◽  
J. H. Williams
Keyword(s):  
Ryanodine Receptor ◽  
Calcium Release ◽  
Binding Capacity ◽  
Polyclonal Antibodies ◽  
Weight Bearing ◽  
Ryanodine Receptors ◽  
Release Mechanism ◽  
Surgical Removal ◽  
Scatchard Analysis ◽  
Western Blots

In the present study, we examined whether weight-bearing condition can regulate the sarcoplasmic reticulum (SR) Ca(2+)-release mechanism. Measurements of alpha 1-subunit dihydropyridine (alpha 1-DHP) and ryanodine receptor levels were made in hypertrophied fast-twitch plantaris muscles 5 wk after surgical removal of synergist muscles (increased weight bearing) and in atrophied slowtwitch soleus muscles (14 days of non-weight bearing) of the rat. Rates of AgNO3-induced SR Ca2+ release were measured with fura 2 as the Ca2+ indicator and pyrophosphate as the precipitating ion during vesicular Ca2+ loading. Ca(2+)-release rates were 38, 49, and 58% lower in vesicles from hypertrophied vs. control muscles at AgNO3 concentrations of 0.05, 0.5, and 5 microM, respectively (control = 18.2 +/- 1.4 microM.mg-1. min-1). Western blots showed no differences in the relative expression of alpha 1-DHP or ryanodine receptor when IIID5 (monoclonal) or GP3 (polyclonal) antibodies were used. There was also no difference in ryanodine (10 nM) binding in Ca(2+)-incubated SR vesicles from hypertrophied muscles, suggesting no difference in the number of channels. In contrast, expression of alpha 1-DHP and ryanodine receptors was increased by 144 and 157% in non-weight-bearing soleus muscles, respectively. Scatchard analysis of DHP binding showed a 40% increase in maximum binding capacity and no change in the dissociation constant with non-weight-bearing muscles. The direction of modification of the SR Ca(2+)-release mechanism is opposite with increased and decreased weight bearing, but the mechanism by which this is achieved appears to be different.

Effects of progesterone induced postmaturity on CBG and pituitary-adrenal activities in the rat foetus

1986 ◽  
Vol 112 (3) ◽  
pp. 396-403 ◽  
Author(s):  
Jean Paul Dupouy ◽  
Alain Chatelain
Keyword(s):  
Binding Capacity ◽  
Sharp Decrease ◽  
Plasma Corticosterone ◽  
Corticosterone Concentration ◽  
Rat Pups ◽  
Body Weights ◽  
Normal Rat ◽  
Severe Jaundice ◽  
High Level ◽  
Subsequent Rise

Abstract. CBG and pituitary-adrenal activities were investigated in intact rat foetuses, in newborns spontaneously delivered by vaginal way and in postmature foetuses from mothers with delayed parturition caused by daily progesterone injection from day 20 of gestation. The postmature foetuses had lower body weights and higher adrenal weights on day 22, 23 and 24 of gestation than newborns of the same conceptional age. The corticosterone binding capacity of the plasma as well as the binding capacity of CBG for corticosterone decreased in intact foetuses for the last 3 days of gestation and stayed very low in pups from day 0 to day 8 postpartum. These parameters decreased more slowly in postmature foetuses; however, the differences between the latter and intact foetuses or newborns were not statistically significant. Similar evolution occurred in intact pregnant and suckling females as well as in females with prolonged gestation. The fall in CBG activity in normal rat pups and the subsequent rise in free steroids could explain a sharp decrease in plasma ACTH levels as well as the drop in adrenal and plasma corticosterone concentration. In foetuses with prolonged gestation, the same phenomenon did not occur. Stress conditions produced by maintaining growing foetuses in utero and the development of severe jaundice maintained high ACTH levels. In contrast, the fall in adrenal and plasma corticosterone concentrations in spite of the high level of circulating ACTH could be mainly due to the progesterone inhibition of the steroidogenic activity of the foetal adrenals.

scholarly journals Fitness Costs of Chlorantraniliprole Resistance Related to the SeNPF Overexpression in the Spodoptera exigua (Lepidoptera: Noctuidae)

2021 ◽  
Vol 22 (9) ◽  
pp. 5027
Author(s):  
Changwei Gong ◽  
Xinge Yao ◽  
Qunfang Yang ◽  
Xuegui Wang ◽  
Yuming Zhang ◽  
...  
Keyword(s):  
Insect Pest ◽  
Ryanodine Receptors ◽  
Oviposition Rate ◽  
Fitness Cost ◽  
Fitness Costs ◽  
Male Adult ◽  
Mating Response ◽  
Transmembrane Segments ◽  
High Level ◽  
Lepidoptera Noctuidae

Spodopteraexigua, a multifeeding insect pest, has developed a high level of resistance to chlorantraniliprole, which is a benzoylurea insecticide that targets the ryanodine receptors (RyRs). Herein, the resistant strain (SE-Sel) and sensitive strain (SE-Sus) were obtained by bidirectional screening for six generations. The potential oviposited eggs and oviposition rate of the SE-Sel strain were dramatically lower than those of the SE-Sus strain; on the contrary, the weights of prepupae and preadult were significantly increased. As a post-mating response, the higher number of non-oviposited eggs in the SE-Sel strain was caused by a lower mating rate. In addition, the expression levels of vitellogenin (SeVg) and its receptor (SeVgR) in the SE-Sel strain were consistently lower than those in the SE-Sus strain. An RyRI4743M mutation, contributing to the resistance to chlorantraniliprole, was located in the S3 transmembrane segments and might have affected the release of calcium ions; it led to the upregulated expression of the neuropeptide SeNPF and its receptor SeNPFR, and the mating and oviposition rate were significantly recovered when the SeNPF was knocked down though RNA interference (RNAi) in the male adult of the SE-Sel strain. Moreover, the expression of the juvenile hormone-binding proteins SeJHBWDS3 and SeJHBAN in the male adult of the SE-Sel strain was significantly decreased, which proved the existence of a fitness cost from another angle. Therefore, these results indicate that the fitness cost accompanied by chlorantraniliprole resistance in S. exigua may be related to the decrease in mating desire due to SeNPF overexpression.

scholarly journals The transcription factor OpWRKY2 positively regulates the biosynthesis of the anticancer drug camptothecin in Ophiorrhiza pumila

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Xiaolong Hao ◽  
Chenhong Xie ◽  
Qingyan Ruan ◽  
Xichen Zhang ◽  
Chao Wu ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anticancer Activity ◽  
Time Course ◽  
Indole Alkaloid ◽  
Fold Increase ◽  
Wrky Transcription Factor ◽  
Mobility Shift ◽  
Pathway Gene ◽  
Low Concentrations ◽  
Encoding Gene

AbstractThe limited bioavailability of plant-derived natural products with anticancer activity poses major challenges to the pharmaceutical industry. An example of this is camptothecin, a monoterpene indole alkaloid with potent anticancer activity that is extracted at very low concentrations from woody plants. Recently, camptothecin biosynthesis has been shown to become biotechnologically amenable in hairy-root systems of the natural producer Ophiorrhiza pumila. Here, time-course expression and metabolite analyses were performed to identify novel transcriptional regulators of camptothecin biosynthesis in O. pumila. It is shown here that camptothecin production increased over cultivation time and that the expression pattern of the WRKY transcription factor encoding gene OpWRKY2 is closely correlated with camptothecin accumulation. Overexpression of OpWRKY2 led to a more than three-fold increase in camptothecin levels. Accordingly, silencing of OpWRKY2 correlated with decreased camptothecin levels in the plant. Further detailed molecular characterization by electrophoretic mobility shift, yeast one-hybrid and dual-luciferase assays showed that OpWRKY2 directly binds and activates the central camptothecin pathway gene OpTDC. Taken together, the results of this study demonstrate that OpWRKY2 acts as a direct positive regulator of camptothecin biosynthesis. As such, a feasible strategy for the over-accumulation of camptothecin in a biotechnologically amenable system is presented.

scholarly journals Complete heart block in cardiac sarcoidosis reversed by corticosteroid therapy: time course of resolution

2021 ◽  
Vol 14 (3) ◽  
pp. e240834
Author(s):  
Anna Tomdio ◽  
Huzaefah Syed ◽  
Kenneth Ellenbogen ◽  
Jordana Kron
Keyword(s):  
Heart Block ◽  
Time Course ◽  
Cardiac Sarcoidosis ◽  
Complete Heart Block ◽  
Positron Emission ◽  
Av Conduction ◽  
Magnetic Resonance Imaging Mri ◽  
High Level ◽  
Active Inflammation ◽  
Lateral Walls

A 53-year-old man was admitted for recurrent syncope and found to have complete heart block (CHB). Cardiac magnetic resonance imaging MRI) showed extensive patchy late gadolinium enhancement in the apical and lateral walls, consistent with cardiac sarcoidosis (CS) but no scar in the septum. A fluorodeoxyglucose (FDG)–positron emission tomography showed FDG uptake in the septum and basal lateral walls. Imaging suggested active inflammation in the septum affecting atrioventricular (AV) conduction but no irreversible fibrosis. Diagnosis of isolated CS requires a high level of suspicion and multidisciplinary teamwork involving heart failure specialists, electrophysiologists and rheumatologists. After specialist and patient discussion, treatment of the disease was initiated with prednisone 40 mg daily, 11 months after presenting with CHB. Three weeks later, ECG with pacing inhibited showed second-degree AV block Mobitz type II and 4 weeks later, AV conduction recovery. This highlights the importance of immediate therapy in reversing AV conduction abnormalities in CS.

scholarly journals The Rapid Extraction of Gist—Early Neural Correlates of High-level Visual Processing

2012 ◽  
Vol 24 (2) ◽  
pp. 521-529 ◽  
Author(s):  
Frank Oppermann ◽  
Uwe Hassler ◽  
Jörg D. Jescheniak ◽  
Thomas Gruber
Keyword(s):  
Visual Processing ◽  
Time Course ◽  
Occipital Cortex ◽  
Stimulus Onset ◽  
Gamma Band ◽  
Oscillatory Activity ◽  
Rapid Extraction ◽  
The Right ◽  
High Level ◽  
Individual Objects

The human cognitive system is highly efficient in extracting information from our visual environment. This efficiency is based on acquired knowledge that guides our attention toward relevant events and promotes the recognition of individual objects as they appear in visual scenes. The experience-based representation of such knowledge contains not only information about the individual objects but also about relations between them, such as the typical context in which individual objects co-occur. The present EEG study aimed at exploring the availability of such relational knowledge in the time course of visual scene processing, using oscillatory evoked gamma-band responses as a neural correlate for a currently activated cortical stimulus representation. Participants decided whether two simultaneously presented objects were conceptually coherent (e.g., mouse–cheese) or not (e.g., crown–mushroom). We obtained increased evoked gamma-band responses for coherent scenes compared with incoherent scenes beginning as early as 70 msec after stimulus onset within a distributed cortical network, including the right temporal, the right frontal, and the bilateral occipital cortex. This finding provides empirical evidence for the functional importance of evoked oscillatory activity in high-level vision beyond the visual cortex and, thus, gives new insights into the functional relevance of neuronal interactions. It also indicates the very early availability of experience-based knowledge that might be regarded as a fundamental mechanism for the rapid extraction of the gist of a scene.

scholarly journals The role of bound potassium ions in the hydrolysis of low concentrations of adenosine triphosphate by preparations of membrane fragments from ox brain cerebral cortex

1970 ◽  
Vol 120 (1) ◽  
pp. 15-24 ◽  
Author(s):  
P. S. G. Goldfarb ◽  
R. Rodnight
Keyword(s):  
Potassium Chloride ◽  
Magnesium Chloride ◽  
Adenosine Triphosphatase ◽  
Time Course ◽  
Atp Hydrolysis ◽  
Protein Ratio ◽  
Low Concentrations ◽  
Hydrolysis Of ◽  
Emission Flame

1. The intrinsic Na+, K+, Mg2+ and Ca2+ contents of a preparation of membrane fragments from ox brain were determined by emission flame photometry. 2. Centrifugal washing of the preparation with imidazole-buffered EDTA solutions decreased the bound Na+ from 90±20 to 24±12, the bound K+ from 27±3 to 7±2, the bound Mg2+ from 20±2 to 3±1 and the bound calcium from 8±1 to <1nmol/mg of protein. 3. The activities of the Na++K++Mg2+-stimulated adenosine triphosphatase and the Na+-dependent reaction forming bound phosphate were compared in the unwashed and washed preparations at an ATP concentration of 2.5μm (ATP/protein ratio 12.5pmol/μg). 4. The Na+-dependent hydrolysis of ATP as well as the plateau concentration of bound phosphate and the rate of dephosphorylation were decreased in the washed preparation. The time-course of formation and decline of bound phosphate was fully restored by the addition of 2.5μm-magnesium chloride and 2μm-potassium chloride. Addition of 2.5μm-magnesium chloride alone fully restored the plateau concentration of bound phosphate, but the rate of dephosphorylation was only slightly increased. Na+-dependent ATP hydrolysis was partly restored with 2.5μm-magnesium chloride; addition of K+ in the range 2–10μm-potassium chloride then further restored hydrolysis but not to the control rate. 5. Pretreatment of the washed preparation at 0°C with 0.5nmol of K+/mg of protein so that the final added K+ in the reaction mixture was 0.1μm restored the Na+-dependent hydrolysis of ATP and the time-course of the reaction forming bound phosphate. 6. The binding of [42K]potassium chloride by the washed membrane preparation was examined. Binding in a solution containing 10nmol of K+/mg of protein was linear over a period of 20min and was inhibited by Na+. Half-maximal inhibition of 42K+-binding required a 100-fold excess of sodium chloride. 7. It was concluded (a) that a significant fraction of the apparent Na+-dependent hydrolysis of ATP observed in the unwashed preparation is due to activation by bound K+ and Mg2+ of the Na++K++Mg2+-stimulated adenosine triphosphatase system and (b) that the enzyme system is able to bind K+ from a solution of 0.5μm-potassium chloride.

Pattern differences in experimental fevers induced by endotoxin, endogenous pyrogen, and prostaglandins

1988 ◽  
Vol 254 (4) ◽  
pp. R633-R640 ◽  
Author(s):  
A. Morimoto ◽  
T. Nakamori ◽  
T. Watanabe ◽  
T. Ono ◽  
N. Murakami
Keyword(s):  
Time Course ◽  
Interleukin 1 ◽  
Bacterial Endotoxin ◽  
Endogenous Pyrogen ◽  
Prolonged Time ◽  
Fever Onset ◽  
Long Latency ◽  
Low Concentrations ◽  
High Concentrations ◽  
Experimentally Induced

To distinguish pattern differences in experimentally induced fevers, we investigated febrile responses induced by intravenous (IV), intracerebroventricular (ICV), and intra-preoptic/anterior hypothalamic (POA) administration of bacterial endotoxin (lipopolysaccharide, LPS), endogenous pyrogen (EP), human recombinant interleukin-1 alpha (IL-1), and prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha). Intravenous LPS, EP, or IL-1 in high concentrations caused biphasic fever. In low concentrations, they induced only the first phase of fever. Latency to onset and time to first peak of fever induced by IV injection of LPS or EP were almost the same as those after ICV or POA injection of PGE2. Fever induced by ICV or POA administration of LPS, EP, IL-1, or PGF2 alpha had a long latency to onset and a prolonged time course. There were significant differences among the latencies to fever onset exhibited by groups that received ICV or POA injections of LPS, EP, or PGF2 alpha and by groups given IV injections of LPS or EP and ICV or POA injections of PGE2. Present observations indicate different patterns of fever produced by several kinds of pyrogens when given by various routes. These results permit us to consider the possibility that there are several mediators or multiprocesses underlying the pathogenesis of fever.

scholarly journals Pattern of pyruvate kinase isozymes in erythroleukemia cell lines and in normal human erythroblasts

Blood ◽  
1984 ◽  
Vol 64 (4) ◽  
pp. 930-936 ◽  
Author(s):  
I Max-Audit ◽  
U Testa ◽  
D Kechemir ◽  
M Titeux ◽  
W Vainchenker ◽  
...  
Keyword(s):  
Pyruvate Kinase ◽  
Cell Lines ◽  
Fetal Liver ◽  
Erythroid Cells ◽  
Low Concentrations ◽  
Erythroleukemia Cell ◽  
Erythroleukemic Cell ◽  
High Level

To further investigate the erythroid nature of the two human erythroleukemia cell lines, K562 and HEL-60, and to define the ontogeny of pyruvate kinase (PK) isozymes (R, M2) in developing human erythroid cells, we have studied the isozymic alterations, if any, during differentiation of these cell lines in vitro and normoblasts isolated from fetal liver in vivo. PK activity of erythroleukemic cell lines was intermediate between that observed in leukocytes and in fetal liver erythroblasts. These cell lines contained a high level of M2-PK, but R- PK was always present, albeit at low concentrations, in all the clones or subclones we studied. Erythroblasts from fetal liver were separated according to density on a Stractan gradient. R-PK levels were nearly constant in the different fractions, whereas M2-PK levels markedly decreased as the erythroblasts became mature and almost completely disappeared in late erythroid cells. Thus, these results clearly demonstrate the erythroid origin of these cell lines.

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