Capacitation of mouse spermatozoa. II. Protein tyrosine phosphorylation and capacitation are regulated by a cAMP-dependent pathway

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1139-1150 ◽  
Author(s):  
P.E. Visconti ◽  
G.D. Moore ◽  
J.L. Bailey ◽  
P. Leclerc ◽  
S.A. Connors ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Tyrosine Phosphorylation ◽  
Bovine Serum ◽  
Biologically Active ◽  
Dependent Manner ◽  
Protein Tyrosine Phosphorylation ◽  
Concentration Dependent Manner ◽  
Mouse Sperm ◽  
Protein Tyrosine

In the accompanying report (Visconti, P.E., Bailey, J.L., Moore, G.D., Pan, D., Olds-Clarke, P. and Kopf, G.S. (1995) Development, 121, 1129–1137) we demonstrated that the tyrosine phosphorylation of a subset of mouse sperm proteins of M(r) 40,000-120,000 was correlated with the capacitation state of the sperm. The mechanism by which protein tyrosine phosphorylation is regulated in sperm during this process is the subject of this report. Cauda epididymal sperm, when incubated in media devoid of NaHCO3, CaCl2 or bovine serum albumin do not display the capacitation-associated increases in protein tyrosine phosphorylation of this subset of proteins. This NaHCO3, CaCl2 or bovine serum albumin requirement for protein tyrosine phosphorylation can be completely overcome by the addition of biologically active, but not inactive, cAMP analogues. Addition of the active cAMP analogues to sperm incubated in media devoid of NaHCO3, CaCl2 or bovine serum albumin overcomes the inability of these media to support capacitation, as assessed by the ability of the cells to acquire the pattern B chlortetracycline fluorescence, to undergo the zona pellucida-induced acrosome reaction and, in some cases, to fertilize metaphase II-arrested eggs in vitro. The effects of the cAMP analogues to enhance protein tyrosine phosphorylation and to promote capacitation appears to be at the level of the cAMP-dependent protein kinase (PKA), since two specific inhibitors of this enzyme (H-89 and Rp-cAMPS) block the capacitation-dependent increases in protein tyrosine phosphorylation in sperm incubated in media supporting capacitation. Capacitation, as assessed by the aforementioned endpoints, also appears to be inhibited by H-89 in a concentration-dependent manner. These results provide further evidence for the interrelationship between protein tyrosine phosphorylation and the appearance of the capacitated state in mouse sperm. They also demonstrate that both protein tyrosine phosphorylation and capacitation appear to be regulated by cAMP/PKA. Up-regulation of protein tyrosine phosphorylation by cAMP/PKA in sperm is, to our knowledge, the first demonstration of such an interrelationship between tyrosine kinase/phosphatase and PKA signaling pathways.

Capacitation of mouse spermatozoa. I. Correlation between the capacitation state and protein tyrosine phosphorylation

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1129-1137 ◽  
Author(s):  
P.E. Visconti ◽  
J.L. Bailey ◽  
G.D. Moore ◽  
D. Pan ◽  
P. Olds-Clarke ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Tyrosine Phosphorylation ◽  
Bovine Serum ◽  
Female Reproductive Tract ◽  
Dependent Manner ◽  
Protein Tyrosine Phosphorylation ◽  
Sperm Capacitation ◽  
Protein Tyrosine

The molecular basis of mammalian sperm capacitation, defined functionally as those processes that confer on the sperm the acquisition of fertilization-competence either in vivo in the female reproductive tract or in vitro, is poorly understood. We demonstrate here that capacitation of caudal epididymal mouse sperm in vitro is accompanied by a time-dependent increase in the protein tyrosine phosphorylation of a subset of proteins of M(r) 40,000-120,000. Incubation of sperm in media devoid of bovine serum albumin, CaCl2 or NaHCO3, components which individually are required for capacitation, prevent the sperm from undergoing capacitation as assessed by the ability of the cells to acquire the pattern B chlortetracycline fluorescence, to undergo the zona pellucida-induced acrosome reaction and, in some cases, to fertilize metaphase II-arrested eggs in vitro. In each of these cases the protein tyrosine phosphorylation of the subset of capacitation-associated proteins does not occur. Protein tyrosine phosphorylation of these particular proteins, as well as sperm capacitation, can be recovered in media devoid of each of these three constituents (bovine serum albumin, CaCl2 or NaHCO3) by adding back the appropriate component in a concentration-dependent manner. The requirement of NaHCO3 for these phosphorylations is not due to an alkalinization of intracellular sperm pH or to an increase in media pH. Caput epididymal sperm, which lack the ability to undergo capacitation in vitro, do not display this capacitation-dependent subset of tyrosine phosphorylated proteins in complete media even after extended incubation periods, and do not fertilize metaphase II-arrested eggs in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)

Protein-Tyrosine Phosphorylation and p72 syk Activation in Human Platelets Stimulated with Collagen Is Dependent upon Glycoprotein Ia/IIa and Actin Polymerization

1996 ◽  
Vol 75 (04) ◽  
pp. 648-654 ◽  
Author(s):  
Naoki Asazuma ◽  
Yutaka Yatomi ◽  
Yukio Ozaki ◽  
Ruomei Qi ◽  
Kenji Kuroda ◽  
...  
Keyword(s):  
Tyrosine Phosphorylation ◽  
Actin Polymerization ◽  
Human Platelets ◽  
Dependent Manner ◽  
Protein Tyrosine Phosphorylation ◽  
Concentration Dependent Manner ◽  
Protein Tyrosine ◽  
Collagen Receptors ◽  
Fibrinogen Binding ◽  
Adp Release

SummaryIn human platelets treated with acetylsalicylic acid, collagen induced protein-tyrosine-phosphorylation of several proteins. The major 75 kDa band included cortactin and autophosphorylated p72 syk . p72 syk activity rapidly increased upon collagen stimulation, whereas p60c-src activation was below detectable levels. A combination of inhibitors to remove the effects of extracellular and intracellular Ca2+, released ADP, and fibrinogen binding to GPIIb/IIIa delayed and attenuated the major 75 kDa band. By contrast, p72 syk activation was not inhibited by these treatments. Cytochalasin D completely inhibited protein tyrosine phosphorylation and p72 syk activation. It also potently inhibited aggregation and [Ca2+]i elevation. Anti-GPMIa/IIa MoAb in a concentration-dependent manner partially attenuated protein tyrosine phosphorylation and p72 syk activation. Its inhibitory effects on intracellular Ca2+ mobilization, release of intracellular granule contents, and aggregation also were partial. No tyrosine kinase activity was coprecipitated with GPIa/IIa. These results suggest that p72 syk activation lies upstream of protein tyrosine phosphorylation, Ca2+ mobilization, ADP release, thromboxane A2 production and aggregation. GPIa/IIa plays a key role in p72 syk activation induced by collagen, but other collagen receptors may work in synergy to fully activate p72 syk . Actin polymerization is a prerequisite for both p72 syk activation and other intracellular signal transduction pathways.

scholarly journals Peroxynitrite modulates tyrosine phosphorylation and phosphoinositide signalling in human neuroblastoma SH-SY5Y cells: attenuated effects in human 1321N1 astrocytoma cells

1998 ◽  
Vol 331 (2) ◽  
pp. 599-606 ◽  
Author(s):  
Xiaohua LI ◽  
Patrizia De SARNO ◽  
Ling SONG ◽  
Joseph S. BECKMAN ◽  
Richard S. JOPE
Keyword(s):  
Tyrosine Phosphorylation ◽  
Phosphoinositide Hydrolysis ◽  
Glutathione Depletion ◽  
Dependent Manner ◽  
Protein Tyrosine Phosphorylation ◽  
Concentration Dependent Manner ◽  
Human Neuroblastoma ◽  
Protein Tyrosine ◽  
Astrocytoma Cells ◽  
1321N1 Astrocytoma

Peroxynitrite may contribute to oxidative stress involving neurodegeneration in several disorders, including Alzheimer's disease. As with other reactive oxygen species, peroxynitrite might affect neuronal signalling systems, actions that could contribute to adaptive or deleterious cellular outcomes, but such effects have not previously been studied. To address this issue directly, peroxynitrite (50–500 µM) was administered to human neuroblastoma SH-SY5Y cells to assess its effects on protein tyrosine nitration, phosphoinositide signalling and protein tyrosine phosphorylation. Peroxynitrite rapidly increased the nitrotyrosine immunoreactivity of numerous proteins, primarily in the cytosol. Peroxynitrite inhibited, in a concentration-dependent manner, phosphoinositide hydrolysis stimulated by activation of muscarinic receptors with carbachol and the inhibition was greater after the depletion of cellular glutathione. In comparison, muscarinic receptor-stimulated phosphoinositide hydrolysis in human astrocytoma 1321N1 cells was less vulnerable to inhibition by peroxynitrite either without or with prior depletion of glutathione. There was a large, rapid and reversible increase in the tyrosine phosphorylation of the p120 Src substrate in peroxynitrite-treated SH-SY5Y cells, a response that was potentiated by glutathione depletion; in contrast, peroxynitrite decreased the tyrosine phosphorylation of focal adhesion kinase and paxillin. Tyrosine phosphorylation of p120 in 1321N1 astrocytoma cells was less sensitive to modulation by peroxynitrite. Thus alterations in phosphoinositide signalling and protein tyrosine phosphorylation were greater in neuroblastoma than astrocytoma cells, and modulation of these signalling processes probably contributes to neuronal mechanisms of the response to peroxynitrite.

scholarly journals β-glucan: Immune boosting potential and antioxidant candidate

2020 ◽  
Vol 11 (1) ◽  
pp. 491-496
Author(s):  
Abhishek Kamboj ◽  
Aanchal Jain ◽  
Tanya Singh ◽  
Ajam Shaikh ◽  
Amit Gupta
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Specific Protein ◽  
Antioxidant Potential ◽  
Cell Culture Supernatant ◽  
Dependent Manner ◽  
Dpph Assay ◽  
Concentration Dependent Manner ◽  
Ifn Gamma

In literature, β-glucan is abundantly found in oats and currently used as well as tested against various varieties of food products. The objective of our study is to evaluate its immune-boosting potential of β-glucan extracted from oats and tested against specific protein antigen i.e., bovine serum albumin (BSA) and also measure its antioxidant activity. For these studies, β-glucan using variable concentration and tested in human whole blood samples (exposed with an optimized concentration of bovine serum albumin, BSA) and examined its immune-boosting potential. In contrast, estimation of Th1 (IFN-gamma) and Th2 (IL-4) cytokines were also measured from cell culture supernatant and also measured its antioxidant potential through DPPH assay. The results showed that β-glucan enhanced immune-boosting activity in a concentration-dependent manner against BSA. In contrast, there is enhancement in Th2 (IL-4) cytokines in a dose-dependent manner, but there is no significant effect in Th1 (IFN-gamma) as compared to control. In addition, β-glucan showed drastic enhanced antioxidant potential, which is confirmed through DPPH assay. The data demonstrate that β-glucan showing immune-boosting properties and antioxidant potential.

Bovine Serum Albumin Enhances Silver Nanoparticle Dissolution Kinetics in a Size- and Concentration-Dependent Manner

Langmuir ◽  
2020 ◽  
Vol 36 (4) ◽  
pp. 1053-1061 ◽  
Author(s):  
Daniel J. Boehmler ◽  
Zachary J. O’Dell ◽  
Christopher Chung ◽  
Kathryn R. Riley
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Silver Nanoparticle ◽  
Bovine Serum ◽  
Dissolution Kinetics ◽  
Dependent Manner ◽  
Concentration Dependent Manner

scholarly journals Constitutive secretion of serum albumin requires reversible protein tyrosine phosphorylation events in trans-Golgi

2005 ◽  
Vol 289 (3) ◽  
pp. C748-C756 ◽  
Author(s):  
Rachel J. Webb ◽  
Jacob D. Judah ◽  
Lee-Chiang Lo ◽  
Geraint M. H. Thomas
Keyword(s):  
Serum Albumin ◽  
Tyrosine Phosphorylation ◽  
Tyrosine Kinases ◽  
Kinase Inhibitor ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Protein Tyrosine Phosphorylation ◽  
Albumin Secretion ◽  
Protein Tyrosine ◽  
Constitutive Secretion

Serum albumin secretion from rat hepatocytes proceeds via the constitutive pathway. Although much is known about the role of protein tyrosine phosphorylation in regulated secretion, nothing is known about its function in the constitutive process. Here we show that albumin secretion is inhibited by the tyrosine kinase inhibitor genistein but relatively insensitive to subtype-selective inhibitors or treatments. Secretion is also blocked in a physiologically identical manner by the tyrosine phosphatase inhibitors pervanadate and bisperoxo(1,10-phenanthroline)-oxovanadate. Inhibition of either the kinase(s) or phosphatase(s) leads to the accumulation of albumin between the trans-Golgi and the plasma membrane, whereas the immediate precursor proalbumin builds up in a proximal compartment. The trans-Golgi marker TGN38 is rapidly dispersed under conditions that inhibit tyrosine phosphatase action, whereas the distribution of the cis-Golgi marker GM130 is insensitive to genistein or pervanadate. By using a specifically reactive biotinylation probe, we detected protein tyrosine phosphatases in highly purified rat liver Golgi membranes. These membranes also contain both endogenous tyrosine kinases and their substrates, indicating that enzymes and substrates for reversible tyrosine phosphorylation are normal membrane-resident components of this trafficking compartment. In the absence of perturbation of actin filaments and microtubules, we conclude that reversible protein tyrosine phosphorylation in the trans-Golgi network is essential for albumin secretion and propose that the constitutive secretion of albumin is in fact a regulated process.

Role of protein tyrosine phosphorylation in H2O2-induced activation of endothelial cell phospholipase D

1996 ◽  
Vol 271 (3) ◽  
pp. L400-L408 ◽  
Author(s):  
V. Natarajan ◽  
S. Vepa ◽  
R. S. Verma ◽  
W. M. Scribner
Keyword(s):  
Endothelial Cells ◽  
Tyrosine Kinase ◽  
Tyrosine Phosphorylation ◽  
Tyrosine Kinase Inhibitors ◽  
Phospholipase D ◽  
Kinase Inhibitors ◽  
Tyrosine Phosphatase ◽  
Dependent Manner ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine

Oxidant-induced activation of phospholipase D (PLD) in bovine pulmonary artery endothelial cells (BPAEC) is independent of protein kinase C and calcium. In the present study, the effects of tyrosine kinase and protein tyrosine phosphatase (PTPase) inhibitors on hydrogen peroxide (H2O2)-induced PLD activation and protein tyrosine phosphorylation were examined in BPAEC. Pretreatment of BPAEC with putative tyrosine kinase inhibitors genistein, tyrphostin, and herbimycin attenuated H2O2 (1 mM)-induced PLD activation. The inhibitory effect of the tyrosine kinase inhibitors was highly specific for H2O2-induced modulation and showed no effect on PLD activation mediated by 12-O-tetradecanoylphorbol 13-acetate or bradykinin. Furthermore, addition of H2O2 increased in a time-dependent manner tyrosine phosphorylation of several proteins (17-200 kDa), as determined by immunoblot analysis with antiphosphotyrosine antibodies. H2O2-mediated protein tyrosine phosphorylation preceded PLD activation, and a good correlation was observed on the effect of genistein in H2O2-induced PLD activation and protein tyrosine phosphorylation. Addition of vanadate, a phosphotyrosine phosphatase inhibitor, synergistically increased both PLD activation and protein tyrosine phosphorylation mediated by H2O2. Moreover, vanadate by itself had minimal effect on basal PLD activity in BPAEC; however, at 10 microM vanadate, an increase in protein tyrosine phosphorylation was observed. In addition to vanadate, phenylarsine oxide and diamide potentiated H2O2-induced PLD activation. These results suggest that tyrosine kinase activation may be involved in H2O2-induced PLD activation in vascular endothelial cells.

scholarly journals Sorbitol Can Fuel Mouse Sperm Motility and Protein Tyrosine Phosphorylation via Sorbitol Dehydrogenase1

2009 ◽  
Vol 80 (1) ◽  
pp. 124-133 ◽  
Author(s):  
Wenlei Cao ◽  
Haig K. Aghajanian ◽  
Lisa A. Haig-Ladewig ◽  
George L. Gerton
Keyword(s):  
Tyrosine Phosphorylation ◽  
Sperm Motility ◽  
Protein Tyrosine Phosphorylation ◽  
Mouse Sperm ◽  
Protein Tyrosine

scholarly journals Human Sperm Remain Motile After a Temporary Energy Restriction but do Not Undergo Capacitation-Related Events

2021 ◽  
Vol 9 ◽  
Author(s):  
Clara I. Marín-Briggiler ◽  
Guillermina M. Luque ◽  
María G. Gervasi ◽  
Natalia Oscoz-Susino ◽  
Jessica M. Sierra ◽  
...  
Keyword(s):  
Tyrosine Phosphorylation ◽  
Metabolic Pathways ◽  
Energy Recovery ◽  
Human Sperm ◽  
Energy Restriction ◽  
Protein Tyrosine Phosphorylation ◽  
Mouse Sperm ◽  
Protein Tyrosine ◽  
Intracellular Ca

To acquire fertilization competence, mammalian sperm must undergo several biochemical and physiological modifications known as capacitation. Despite its relevance, the metabolic pathways that regulate the capacitation-related events, including the development of hyperactivated motility, are still poorly described. Previous studies from our group have shown that temporary energy restriction in mouse sperm enhanced hyperactivation, in vitro fertilization, early embryo development and pregnancy rates after embryo transfer, and it improved intracytoplasmic sperm injection results in the bovine model. However, the effects of starvation and energy recovery protocols on human sperm function have not yet been established. In the present work, human sperm were incubated for different periods of time in medium containing glucose, pyruvate and lactate (NUTR) or devoid of nutrients for the starving condition (STRV). Sperm maintained in STRV displayed reduced percentages of motility and kinematic parameters compared to cells incubated in NUTR medium. Moreover, they did not undergo hyperactivation and showed reduced levels of ATP, cAMP and protein tyrosine phosphorylation. Similar to our results with mouse sperm, starvation induced increased intracellular Ca2+ concentrations. Starved human sperm were capable to continue moving for more than 27 h, but the incubation with a mitochondrial uncoupler or inhibitors of oxidative phosphorylation led to a complete motility loss. When exogenous nutrients were added back (sperm energy recovery (SER) treatment), hyperactivated motility was rescued and there was a rise in sperm ATP and cAMP levels in 1 min, with a decrease in intracellular Ca2+ concentration and no changes in sperm protein tyrosine phosphorylation. The finding that human sperm can remain motile for several hours under starvation due to mitochondrial use of endogenous metabolites implies that other metabolic pathways may play a role in sperm energy production. In addition, full recovery of motility and other capacitation parameters of human sperm after SER suggests that this treatment might be used to modulate human sperm fertilizing ability in vitro.

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