Adenosine 3',5-cyclic monophosphate phosphodiesterase activity in granulosa cells from Booroola × Romney ewes with and without the F gene

1989 ◽  
Vol 120 (2) ◽  
pp. 287-293
Author(s):  
K. P. McNatty ◽  
D. A. Heath ◽  
S. Lun ◽  
N. L. Hudson
Keyword(s):  
Granulosa Cells ◽  
Maximum Velocity ◽  
Camp Accumulation ◽  
Camp Phosphodiesterase ◽  
F Gene ◽  
Specific Effects ◽  
The Mean ◽  
Camp Hydrolysis ◽  
Camp Levels ◽  
In Cells

ABSTRACT Granulosa cells from ovarian follicles (≥ 1 mm diameter) in Booroola ewes which are homozygous (FF) or heterozygous (F+) for the F gene have previously been shown to produce significantly more cAMP in response to FSH or LH than those from similar sized follicles in ewes without the F gene (++). The aim of these studies was to test whether these F gene-specific differences arose because of differences in cAMP-phosphodiesterase (cAMP-PDE) activity. In the first study using 1 μmol cAMP/1 as substrate, no F gene-specific effects were noted in cAMP-PDE activity in granulosa cells from small (1–2·5 mm diameter, n = 4 per genotype) or large (≥ 3 mm diameter, n = 4 per genotype) follicles from FF, F+ or ++ ewes, despite F gene-specific effects in FSH (1 μg/ml)and LH (0·1 μg/ml)-induced cAMP accumulation in these same cell preparations. The overall mean levels of cAMP-PDE across all genotypes in cells from small and large follicles were 0·47± 0·04 (s.e.m., n = 12) and 0·28 ± 0·03 pmol cAMP/106 cells per min respectively; the mean PDE activity in cells from small follicles was significantly (P < 0·05) higher compared with that in cells from large follicles. In a second study, granulosa cells from each genotype were pooled over all follicle sizes (≥ 1 mm diameter, one pool per genotype) and the rates of cAMP hydrolysis tested over a range of substrate concentrations (0–16 μmol/l) but no genespecific differences with respect to the Michaelis constant and maximum velocity were noted. In a third study, the rates of FSH-induced cAMP accumulation by granulosa cells from small and large follicles of FF and ++ ewes were examined in the presence of a PDE inhibitor, 1-methyl-3-isobutylxanthine (0·2 mmol/l). In this study FSH (1 μg/ml) stimulated significantly more cAMP in cells from FF compared with ++ ewes in both small and large follicles. In cells from small follicles, the mean ± s.e.m. FSHstimulated cAMP levels were 4·7 ± 0·8 (n = 6) for FF ewes and 1·8 ± 0·4 (n = 8) pmol/106 cells per h for ++ ewes (P < 0·025). In large follicles, the cAMP levels were 17·0 ± 3·5 (n = 6) for FF ewes and 6·3 ± 0·5 pmol/106 cells per h for ++ ewes (P < 0·05). Collectively, these data suggest that the F genespecific difference in cAMP synthesis is probably the result of an event(s) associated with its formation rather than its degradation. Journal of Endocrinology (1989) 120, 287–293

Gap junction-dependent and -independent EDHF-type relaxations may involve smooth muscle cAMP accumulation

2002 ◽  
Vol 282 (4) ◽  
pp. H1548-H1555 ◽  
Author(s):  
Andrew T. Chaytor ◽  
Hannah J. Taylor ◽  
Tudor M. Griffith
Keyword(s):  
Smooth Muscle ◽  
Gap Junction ◽  
Phosphodiesterase Inhibitor ◽  
Channel Blockers ◽  
Camp Accumulation ◽  
Camp Phosphodiesterase ◽  
Mechanical Responses ◽  
A 23187 ◽  
Extracellular Release ◽  
Camp Levels

We have compared the mechanisms that contribute to endothelium-derived hyperpolarizing factor (EDHF)-type responses induced by ACh and the Ca2+ ionophore A-23187 in the rabbit iliac artery. Relaxations to both agents were associated with ∼1.5-fold elevations in smooth muscle cAMP levels and were attenuated by the adenylyl cyclase inhibitor 2′,5′-dideoxyadenosine (DDA) and potentiated by the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). Mechanical responses were inhibited by coadministration of the Ca2+-activated K+channel blockers apamin and charybdotoxin, both in the absence and presence of IBMX, but were unaffected by blockade of ATP-sensitive K+ channels with the sulphonylurea glibenclamide. Relaxations and elevations in cAMP evoked by ACh were abolished by 18α-glycyrrhetinic acid, which disrupts gap junction plaques, whereas the corresponding responses to A-23187 were unaffected by this agent. Consistently, in “sandwich” bioassay experiments, A-23187, but not ACh, elicited extracellular release of a factor that evoked relaxations that were inhibited by DDA and potentiated by IBMX. These findings provide evidence that EDHF-type relaxations of rabbit iliac arteries evoked by ACh and A-23187 depend on cAMP accumulation in smooth muscle, but involve signaling via myoendothelial gap junctions and the extracellular space, respectively.

Ticlopidine and Clopidogrel (SR 25990C) Selectively Neutralize ADP Inhibition of PGE1-Activated Platelet Adenylate Cyclase in Rats and Rabbits

1991 ◽  
Vol 65 (02) ◽  
pp. 186-190 ◽  
Author(s):  
G Defreyn ◽  
C Gachet ◽  
P Savi ◽  
F Driot ◽  
J P Cazenave ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Adenylate Cyclase ◽  
Direct Effect ◽  
Cyclic Nucleotide ◽  
Camp Accumulation ◽  
Rabbit Platelets ◽  
Camp Levels ◽  
Kinetics Of ◽  
Camp Elevation ◽  
Rat Platelets

SummaryTiclopidine and its potent analogue, clopidogrel, are powerful inhibitors of ADP-induced platelet aggregation. In order to improve the understanding of this ADP-selectivity, we studied the effect of these compounds on PGE1-stimulated adenylate cyclase and on the inhibition of this enzyme by ADP, epinephrine and thrombin. Neither drug changed the basal cAMP levels nor the kinetics of cAMP accumulation upon PGEj-stimulation in rat or rabbit platelets, which excludes any direct effect on adenylate cyclase or on cyclic nucleotide phosphodiesterase. However, the drop in cAMP levels observed after addition of ADP to PGEr stimulated control platelets was inhibited in platelets from treated animals. In contrast, the drop in cAMP levels produced by epinephrine was not prevented by either drug in rabbit platelets. In rat platelets, thrombin inhibited the PGEX-induced cAMP elevation but this effect seems to be entirely mediated by the released ADP. Under these conditions, it was not surprising to find that clopidogrel also potently inhibited that effect of thrombin on platelet adenylate cyclase. In conclusion, ticlopidine and clopidogrel selectively neutralize the ADP inhibition of PGEr activated platelet adenylate cyclase in rats and rabbits.

Synergistic Antiplatelet Effect of Ridogrel, a Combined Thromboxane Receptor Antagonist and Thromboxane Synthase Inhibitor, and UDCG-212, a cAMP-Phosphodiesterase Inhibitor

1993 ◽  
Vol 70 (05) ◽  
pp. 822-825 ◽  
Author(s):  
B Hoet ◽  
J Arnout ◽  
H Deckmyn ◽  
J Vermylen
Keyword(s):  
Arachidonic Acid ◽  
Platelet Aggregation ◽  
Receptor Antagonist ◽  
Phosphodiesterase Inhibitor ◽  
Camp Phosphodiesterase ◽  
Thromboxane Receptor ◽  
Thromboxane Synthase Inhibitor ◽  
Thromboxane Receptor Antagonist ◽  
Synthase Inhibitor ◽  
Camp Levels

SummaryRidogrel, a combined thromboxane receptor antagonist and thromboxane synthase inhibitor (1), inhibits platelet aggregation. Following stimulation with arachidonic acid, cAMP-levels are increased in human platelets preincubated with ridogrel, this is due to the known reorientation of the metabolism of the formed endoperoxides towards adenylate cyclase stimulating prostaglandins.Pretreatment of resting platelets with UDCG-212, a cAMP-phosphodiesterase inhibitor (2), also inhibits platelet aggregation induced by arachidonic acid, concomitant with an increase in cAMP levels, due to an inhibition of its breakdown. Under basal conditions, cAMP also is increased.By combining the two drugs, a more than additive action was observed on platelet aggregation and on both resting and stimulated platelet cAMP content. The appropriate combination may result in a more effective antiplatelet strategy.

Quantitative studies of the reactions to horizontal angular accelerations in axolotls. I. The head-turning reflexes of normal animals

1977 ◽  
Vol 66 (1) ◽  
pp. 1-14
Author(s):  
K. Brandle
Keyword(s):  
Stimulus Intensity ◽  
Time Course ◽  
Maximum Velocity ◽  
Afferent Input ◽  
Head Turning ◽  
Mean Values ◽  
Quantitative Studies ◽  
The Mean ◽  
Negative Exponential ◽  
Total Angle

1. Artifically metamorphosed axolotls were exposed to both brief (impulse) and long-lasting horizontal angular accelerations on a turn-table. The animals responded with a head-turning reaction. 2. The general course of the reaction to impulse acceleration was independent of stimulus intensity. The velocity of the head movement first increased to a maximum exponentially and then decreased in a negative exponential manner. Stimulus intensity had a linear relationship to the mean maximum velocity and mean total angle covered by head-turning. The average velocity-time curves at various stimulus intensities differed only by a velocity factor. 3. During long-lasting constant accelerations the velocity of the head-turning increased to a maximum velocity in a sigmoid time-course and then decreased, first to a constant velocity, and then further. Mean values of the maximum velocity were correlated linearly with the stimulus intensity. 4. It was concluded that the head-turning reflexes in axolotls do not agree with the accepted movements of the vertebrate cupula and therefore are not a simple ‘copy’ of the afferent input. It is also suggested that the reaction threshold differes from that for the labyrinthine input.

Control of cAMP in lung endothelial cell phenotypes. Implications for control of barrier function

1999 ◽  
Vol 277 (1) ◽  
pp. L119-L126 ◽  
Author(s):  
Troy Stevens ◽  
Judy Creighton ◽  
W. Joseph Thompson
Keyword(s):  
Endothelial Cells ◽  
Adenylyl Cyclase ◽  
Barrier Function ◽  
Enzyme Regulation ◽  
Calcium Sensitivity ◽  
Cytosolic Calcium ◽  
Calcium Entry ◽  
Camp Accumulation ◽  
Camp Phosphodiesterase ◽  
Conversion Rates

Pulmonary microvascular endothelial cells (PMVECs) form a more restrictive barrier to macromolecular flux than pulmonary arterial endothelial cells (PAECs); however, the mechanisms responsible for this intrinsic feature of PMVECs are unknown. Because cAMP improves endothelial barrier function, we hypothesized that differences in enzyme regulation of cAMP synthesis and/or degradation uniquely establish an elevated content in PMVECs. PMVECs possessed 20% higher basal cAMP concentrations than did PAECs; however, increased content was accompanied by 93% lower ATP-to-cAMP conversion rates. In PMVECs, responsiveness to β-adrenergic agonist (isoproterenol) or direct adenylyl cyclase (forskolin) activation was attenuated and responsiveness to phosphodiesterase inhibition (rolipram) was increased compared with those in PAECs. Although both types of endothelial cells express calcium-inhibited adenylyl cyclase, constitutive PMVEC cAMP accumulation was not inhibited by physiological rises in cytosolic calcium, whereas PAEC cAMP accumulation was inhibited 30% by calcium. Increasing either PMVEC calcium entry by maximal activation of store-operated calcium entry or ATP-to-cAMP conversion with rolipram unmasked calcium inhibition of adenylyl cyclase. These data indicate that suppressed calcium entry and low ATP-to-cAMP conversion intrinsically influence calcium sensitivity. Adenylyl cyclase-to-cAMP phosphodiesterase ratios regulate cAMP at elevated levels compared with PAECs, which likely contribute to enhanced microvascular barrier function.

scholarly journals Phospholipase Cβ3 Mediates LH-Induced Granulosa Cell Differentiation

Endocrinology ◽  
2011 ◽  
Vol 152 (7) ◽  
pp. 2857-2869 ◽  
Author(s):  
Francesc X. Donadeu ◽  
Cristina L. Esteves ◽  
Lynsey K. Doyle ◽  
Catherine A. Walker ◽  
Stephanie N. Schauer ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Granulosa Cells ◽  
Inositol Phosphate ◽  
Ex Vivo ◽  
Receptor Expression ◽  
Target Cells ◽  
Down Regulation ◽  
Protein Levels ◽  
Prostaglandin Endoperoxide Synthase ◽  
In Cells

Previous studies showed that under certain conditions LH can stimulate not only adenylate cyclase (AC) but also phospholipase Cβ (PLCβ) signaling in target cells; however, the physiological involvement of PLCβ in LH-induced ovarian follicular cell differentiation has not been determined. To address this, ex vivo expression analyses and specific PLCβ targeting were performed in primary bovine granulosa cells. Expression analyses in cells from small (2.0–5.9 mm), medium (6.0–9.9 mm), and ovulatory-size (10.0–13.9 mm) follicles revealed an increase in mRNA and protein levels of heterotrimeric G protein subunits-αs, -αq, -α11, and -αi2 in ovulatory-size follicles, simultaneous with a substantial increase in LH receptor expression. Among the four known PLCβ isoforms, PLCβ3 (PLCB3) was specifically up-regulated in cells from ovulatory-size follicles, in association with a predominantly cytoplasmic location of PLCB3 in these cells and a significant inositol phosphate response to LH stimulation. Furthermore, RNA interference-mediated PLCB3 down-regulation reduced the ability of LH to induce hallmark differentiation responses of granulosa cells, namely transcriptional up-regulation of prostaglandin-endoperoxide synthase 2 and down-regulation of both aromatase expression and estradiol production. Responses to the AC agonist, forskolin, however, were not affected. In addition, PLCB3 down-regulation did not alter cAMP responses to LH in granulosa cells, ruling out a primary involvement of AC in mediating the effects of PLCB3. In summary, we provide evidence of a physiological involvement of PLCβ signaling in ovulatory-size follicles and specifically identify PLCB3 as a mediator of LH-induced differentiation responses of granulosa cells.

scholarly journals Influence of Rigid Emerged Vegetation in a Channel Bend on Bed Topography and Flow Velocity Field: Laboratory Experiments

Water ◽  
2019 ◽  
Vol 12 (1) ◽  
pp. 118 ◽  
Author(s):  
Hossein Hamidifar ◽  
Alireza Keshavarzi ◽  
Paweł M. Rowiński
Keyword(s):  
Laboratory Experiments ◽  
Flow Characteristics ◽  
Maximum Velocity ◽  
Flow Conditions ◽  
Mean Flow ◽  
Bed Topography ◽  
Channel Bend ◽  
Bed Erosion ◽  
Flow Hydraulics ◽  
The Mean

Trees have been used extensively by river managers for improving the river environment and ecology. The link between flow hydraulics, bed topography, habitat availability, and organic matters is influenced by vegetation. In this study, the effect of trees on the mean flow, bed topography, and bed shear stress were tested under different flow conditions. It was found that each configuration of trees produced particular flow characteristics and bed topography patterns. The SR (single row of trees) model appeared to deflect the maximum velocity downstream of the bend apex toward the inner bank, while leading the velocity to be more uniformly distributed throughout the bend. The entrainment of sediment particles occurred toward the area with higher values of turbulent kinetic energy (TKE). The results showed that both SR and DR (double rows of trees) models are effective in relieving bed erosion in sharp ingoing bends. The volume of the scoured bed was reduced up to 70.4% for tests with trees. This study shows the effectiveness of the SR model in reducing the maximum erosion depth.

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