Silymarin and protein kinase A inhibitor modulate glucose-mediated mouse sperm motility: An in vitro study

2015 ◽  
Vol 15 (3) ◽  
pp. 172-177 ◽  
Author(s):  
Yi-Chuan Chen ◽  
Li-Chern Pan ◽  
Cheng-Wei Lai ◽  
Ying-Shan Chien ◽  
Tzu-Hua Wu
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Sperm Motility ◽  
In Vitro Study ◽  
Vitro Study ◽  
Mouse Sperm ◽  
Kinase A

scholarly journals Quantification of Protein Kinase A (PKA) Activity by An in vitro Radioactive Assay Using the Mouse Sperm Derived Enzyme

BIO-PROTOCOL ◽  
2020 ◽  
Vol 10 (12) ◽  
Author(s):  
Cintia Stival ◽  
Carolina Baro Graf ◽  
Pablo Visconti ◽  
Dario Krapf
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Mouse Sperm ◽  
Radioactive Assay ◽  
Kinase A

scholarly journals Activity, expression and function of a second Drosophila protein kinase A catalytic subunit gene.

Genetics ◽  
1995 ◽  
Vol 141 (4) ◽  
pp. 1507-1520 ◽  
Author(s):  
A Meléndez ◽  
W Li ◽  
D Kalderon
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Essential Gene ◽  
Sequence Similarity ◽  
Catalytic Subunit ◽  
Subunit Gene ◽  
Drosophila Protein ◽  
Pka Catalytic Subunit ◽  
Kinase A

Abstract The DC2 gene was isolated previously on the basis of sequence similarity to DC0, the major Drosophila protein kinase A (PKA) catalytic subunit gene. We show here that the 67-kD Drosophila DC2 protein behaves as a PKA catalytic subunit in vitro. DC2 is transcribed in mesodermal anlagen of early embryos. This expression depends on dorsal but on neither twist nor snail activity. DC2 transcriptional fusions mimic this embryonic expression and are also expressed in subsets of cells in the optic lamina, wing disc and leg discs of third instar larvae. A saturation screen of a small deficiency interval containing DC2 for recessive lethal mutations yielded no DC2 alleles. We therefore isolated new deficiencies to generate deficiency trans-heterozygotes that lacked DC2 activity. These animals were viable and fertile. The absence of DC2 did not affect the viability or phenotype of imaginal disc cells lacking DC0 activity or embryonic hatching of animals with reduced DC0 activity. Furthermore, transgenes expressing DC2 from a DC0 promoter did not efficiently rescue a variety of DC0 mutant phenotypes. These observations indicate that DC2 is not an essential gene and is unlikely to be functionally redundant with DC0, which has multiple unique functions during development.

scholarly journals Membrane-Bound Protein Kinase A Inhibits Smooth Muscle Cell Proliferation In Vitro and In Vivo by Amplifying cAMP–Protein Kinase A Signals

2001 ◽  
Vol 88 (3) ◽  
pp. 319-324 ◽  
Author(s):  
Ciro Indolfi ◽  
Eugenio Stabile ◽  
Carmela Coppola ◽  
Adriana Gallo ◽  
Cinzia Perrino ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Smooth Muscle ◽  
Protein Kinase ◽  
Smooth Muscle Cell ◽  
Protein Kinase A ◽  
Membrane Bound ◽  
Proliferation In Vitro ◽  
Kinase A

scholarly journals The cross-regulation of Gi-protein by cholera toxin involves a phosphorylation by protein kinase A

1995 ◽  
Vol 306 (3) ◽  
pp. 765-769 ◽  
Author(s):  
R Levistre ◽  
M Berguerand ◽  
G Bereziat ◽  
J Masliah
Keyword(s):  
Arachidonic Acid ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Cholera Toxin ◽  
Inhibitory Effect ◽  
Negative Control ◽  
Adp Ribosylation ◽  
Gi Proteins ◽  
Kinase A

Pretreatment of alveolar macrophages with cholera toxin inhibits the release of arachidonic acid induced by the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine. The results presented here show that cholera toxin might exert its inhibitory effect through the phosphorylation of Gi alpha by protein kinase A (PKA). (1) Gi-proteins from cells pretreated with cholera toxin showed parallel increases in their sensitivity to ADP-ribosylation by toxins in vitro and in Gi alpha phosphorylation. By contrast, the Gi alpha concentration was unchanged. (2) Cholera toxin pretreatment also decreased the functional activity of Gi, as assessed by the inhibition (80%) of agonist-induced binding of guanosine-5′-[gamma-thio]triphosphate (GTP[gamma S]). (3) These effects of cholera toxin were blocked by a specific PKA inhibitor, N-(2-[methyl-amino]ethyl)-3-isoquinolinesulphonamide dihydrochloride (H8) and mimicked by a cyclic AMP (cAMP) analogue and a phosphatase inhibitor. (4) Gi alpha was also phosphorylated in vitro by the catalytic subunit of PKA. In contrast with other cell systems, the stimulation of protein kinase C seems to have no effect on the sensitivity of Gi to ADP-ribosylation or on its phosphorylation. Therefore, the phosphorylation of Gi-proteins by PKA seems to be the actual target of the negative control of arachidonic acid release via the cAMP-mediated pathway.

scholarly journals Phosphorylation of SOX9 by Cyclic AMP-Dependent Protein Kinase A Enhances SOX9's Ability To Transactivate aCol2a1 Chondrocyte-Specific Enhancer

2000 ◽  
Vol 20 (11) ◽  
pp. 4149-4158 ◽  
Author(s):  
Wendong Huang ◽  
Xin Zhou ◽  
Véronique Lefebvre ◽  
Benoit de Crombrugghe
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Cyclic Amp ◽  
Protein Kinase A ◽  
Chondrocyte Differentiation ◽  
Phosphorylation Sites ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Kinase A

ABSTRACT Sox9 is a high-mobility-group domain-containing transcription factor required for chondrocyte differentiation and cartilage formation. We used a yeast two-hybrid method based on Son of Sevenless (SOS) recruitment to screen a chondrocyte cDNA library and found that the catalytic subunit of cyclic AMP (cAMP)-dependent protein kinase A (PKA-Cα) interacted specifically with SOX9. Next we found that two consensus PKA phosphorylation sites within SOX9 could be phosphorylated by PKA in vitro and that SOX9 could be phosphorylated by PKA-Cα in vivo. In COS-7 cells cotransfected with PKA-Cα and SOX9 expression plasmids, PKA enhanced the phosphorylation of wild-type SOX9 but did not affect phosphorylation of a SOX9 protein in which the two PKA phosphorylation sites (S64 and S211) were mutated. Using a phosphospecific antibody that specifically recognized SOX9 phosphorylated at serine 211, one of the two PKA phosphorylation sites, we demonstrated that addition of cAMP to chondrocytes strongly increased the phosphorylation of endogenous Sox9. In addition, immunohistochemistry of mouse embryo hind legs showed that Sox9 phosphorylated at serine 211 was principally localized in the prehypertrophic zone of the growth plate, corresponding to the major site of expression of the parathyroid hormone-related peptide (PTHrP) receptor. Since cAMP has previously been shown to effectively increase the mRNA levels of Col2a1 and other specific markers of chondrocyte differentiation in culture, we then asked whether PKA phosphorylation could modulate the activity of SOX9. Addition of 8-bromo-cAMP to chondrocytes in culture increased the activity of a transiently transfected SOX9-dependent 48-bp Col2a1chondrocyte-specific enhancer; similarly, cotransfection of PKA-Cα increased the activity of this enhancer. Mutations of the two PKA phosphorylation consensus sites of SOX9 markedly decreased the PKA-Cα activation of this enhancer by SOX9. PKA phosphorylation and the mutations in the consensus PKA phosphorylation sites of SOX9 did not alter its nuclear localization. In vitro phosphorylation of SOX9 by PKA resulted in more efficient DNA binding. We conclude that SOX9 is a target of cAMP signaling and that phosphorylation of SOX9 by PKA enhances its transcriptional and DNA-binding activity. Because PTHrP signaling is mediated by cAMP, our results support the hypothesis that Sox9 is a target of PTHrP signaling in the growth plate and that the increased activity of Sox9 might mediate the effect of PTHrP in maintaining the cells as nonhypertrophic chondrocytes.

scholarly journals Disruption of protein kinase A localization induces acrosomal exocytosis in capacitated mouse sperm

2018 ◽  
Vol 293 (24) ◽  
pp. 9435-9447 ◽  
Author(s):  
Cintia Stival ◽  
Carla Ritagliati ◽  
Xinran Xu ◽  
Maria G. Gervasi ◽  
Guillermina M. Luque ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Mouse Sperm ◽  
Acrosomal Exocytosis ◽  
Kinase A

ROPN1, a Protein Kinase A-Like (R2D2) Protein, Regulates Sperm Motility.

2011 ◽  
Vol 85 (Suppl_1) ◽  
pp. 804-804
Author(s):  
Sarah E. Fiedler ◽  
Daniel W. Carr
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Sperm Motility ◽  
Kinase A
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