scholarly journals Acidic hyaluronidase activity is present in mouse sperm and is reduced in the absence of SPAM1: Evidence for a role for hyaluronidase 3 in mouse and human sperm

2010 ◽  
Vol 77 (9) ◽  
pp. 759-772 ◽  
Author(s):  
Kristen L. Reese ◽  
Rolands G. Aravindan ◽  
Genevieve S. Griffiths ◽  
Minghai Shao ◽  
Yipei Wang ◽  
...  
Keyword(s):  
Human Sperm ◽  
Mouse Sperm ◽  
Hyaluronidase Activity

scholarly journals Cytochemical detection of hyaluronidase activity in single human and mouse sperm by an improved substrate-film technique.

1984 ◽  
Vol 32 (1) ◽  
pp. 63-66 ◽  
Author(s):  
R Waibel ◽  
L C Ginsberg ◽  
G Ficsor
Keyword(s):  
Germ Cells ◽  
Screening Method ◽  
Human Sperm ◽  
Simple Method ◽  
Male Germ Cells ◽  
Mouse Sperm ◽  
Hyaluronidase Activity ◽  
Film Method ◽  
Human And Mouse ◽  
Substrate Film

A substrate-film method is described that allows the detection of hyaluronidase activity in nearly 100% of single human and mouse sperm. The level of hyaluronidase activity as determined by halo diameters was greater in mouse than in human sperm. This simple method may have use as a screening method for identifying compounds that cause developmental or genetic defects in male germ cells, or for the diagnosis of infertility due to decreased hyaluronidase activity.

scholarly journals Soluble adenylyl cyclase inhibition prevents human sperm functions essential for fertilization

2021 ◽  
Author(s):  
Melanie Balbach ◽  
Lubna Ghanem ◽  
Thomas Rossetti ◽  
Navpreet Kaur ◽  
Carla Ritagliati ◽  
...  
Keyword(s):  
Adenylyl Cyclase ◽  
Acrosome Reaction ◽  
Human Sperm ◽  
Comprehensive Analysis ◽  
Multiple Points ◽  
Soluble Adenylyl Cyclase ◽  
Mouse Sperm ◽  
On Demand ◽  
Sperm Functions

AbstractSoluble adenylyl cyclase (sAC: ADCY10) is essential for activating dormant sperm. Studies of freshly dissected mouse sperm identified sAC as needed for initiating capacitation and activating motility. We now use an improved sAC inhibitor, TDI-10229, for a comprehensive analysis of sAC function in human sperm. Unlike dissected mouse sperm, human sperm are collected post-ejaculation, after sAC activity has already been stimulated. Even in ejaculated human sperm, TDI-10229 interrupts stimulated motility and capacitation, and it prevents acrosome reaction in capacitated sperm. At present, there are no non-hormonal, pharmacological methods for contraception. Because sAC activity is required post-ejaculation at multiple points during the sperm’s journey to fertilize the oocyte, sAC inhibitors define candidates for non-hormonal, on-demand contraceptives suitable for delivery via intravaginal devices in females.

139. THE REGULATION OF TRP53 IN SPERM

2009 ◽  
Vol 21 (9) ◽  
pp. 58
Author(s):  
H. Mudaliar ◽  
C. O'Neill
Keyword(s):  
Protein Synthesis ◽  
Western Blot ◽  
Marked Reduction ◽  
Acrosome Reaction ◽  
Human Sperm ◽  
Cell Stress ◽  
Time Dependent ◽  
Mouse Sperm ◽  
Tumour Suppressor Protein ◽  
Acute Increase

TRP53 is a tumour suppressor protein that is a universal sensor of cell stress. Upon ejaculation, sperm undergo a process of capacitation which allows them to become fertile. We have previously shown that mouse and human sperm possess TRP53. In this study we analyzed the regulation TRP53 presence in sperm. Mouse sperm were collected from the epididymides and incubated for various times in fertilisation medium. TRP53 was detected by both Western blot analysis and immunolocalization. We found that sperm collected directly from the epididymis generally had little or no detectable TRP53. The level increased with time in culture over a period of 120 min. Most of the TRP53 was detected in the midpiece of sperm although it was also detected in the head of a small proportion of sperm. The increase in TRP53 with time accompanied the increase in the proportion of sperm undergoing the acrosome reaction. Yet, Trp53-null sperm still underwent the acrosome reaction at a normal rate. By contrast, sperm that were prevented from undergoing capacitation and the acrosome reaction (by the removal of albumin or calcium from media) showed a marked reduction in the amount of TRP53 detected. This shows that TRP53 may be dependent upon capacitation, but the reverse was not the case. Inhibition of protein synthesis by puromycin did not block the time-dependent increase in TRP53 in sperm. Canonically, the TRP53 level is controlled by its rate of degradation by MDM2-ubiquitin mediated proteolysis. We found that MDM2 was present in sperm and inhibition of MDM2 (by Nutlin-3) caused an acute increase in TRP53 detected in sperm. This study shows that TRP53 levels are acutely regulated in sperm during the time that sperm acquire the capacity to fertilise, yet sperm lacking TRP53 are capable of fertility. Identification of the role for this TRP53 in sperm is under investigation.

ZP2 peptide beads that select human sperm in vitro, decoy mouse sperm in vivo, and provide reversible contraception

2016 ◽  
Vol 106 (3) ◽  
pp. e112
Author(s):  
M. Avella ◽  
B. Baibakov ◽  
A. Burkart Sadusky ◽  
J. Dean
Keyword(s):  
Human Sperm ◽  
Mouse Sperm

scholarly journals A mouse sperm decapacitation factor receptor is phosphatidylethanolamine-binding protein 1

Reproduction ◽  
2005 ◽  
Vol 130 (4) ◽  
pp. 497-508 ◽  
Author(s):  
Rachel Gibbons ◽  
Susan A Adeoya-Osiguwa ◽  
Lynn R Fraser
Keyword(s):  
Plasma Membrane ◽  
Binding Protein ◽  
Human Sperm ◽  
Surface Proteins ◽  
Mouse Sperm ◽  
Fertility Treatments ◽  
State Dependent ◽  
Sperm Plasma Membrane ◽  
Associated Proteins ◽  
Functional Consequences

Capacitation is a pivotal event for mammalian spermatozoa, involving the loss of surface proteins known as decapacitation factors (DF) and consequent acquisition of fertilizing ability. Earlier studies showed that a mouse sperm DF binds to a receptor, DF-R, whose attachment to the sperm plasma membrane appears to involve a glycosylphosphatidylinositol (GPI) anchor. In the present study, purification and subsequent sequencing of DF-R has identified this ~23 kDa protein as phosphatidyletha-nolamine-binding protein 1 (PEBP 1). To obtain functional evidence that supports sequence homology data, purified recombinant PEBP 1 and PEBP 2 were evaluated for biological activity. While PEBP 1 was able to remove DF activity in solution at concentrations above ~1 nmol/l, PEBP 2 was ineffective, even at 600 nmol/l; this confirmed that DF-R is PEBP 1. Anti-PEBP 1 antiserum recognized recombinant PEBP 1 and a ~23 kDa protein in both mouse and human sperm lysates. Immunolocalization studies revealed that DF-R/PEBP 1 is located on the acrosomal cap, the post-acrosomal region and the flagellum of both mouse and human spermatozoa, with epitope accessibility being capacitation state-dependent and reversible. Treatment of cells with a phospholipase able to cleave GPI anchors essentially abolished immunostaining, thus confirming the extracellular location of DF-R/PEBP 1. We suggest that DF-R/PEBP 1 plays its fundamental role in capacitation by causing alterations in the sperm plasma membrane in both head and flagellum, with functional consequences for membrane-associated proteins. Obtaining more detail about DF ↔ DF-R interactions could lead to useful applications in both fertility treatments and new contraceptive approaches.

scholarly journals Conserved Mechanism of Bicarbonate-Induced Sensitization of CatSper Channels in Human and Mouse Sperm

2021 ◽  
Vol 9 ◽  
Author(s):  
Juan J. Ferreira ◽  
Pascale Lybaert ◽  
Lis C. Puga-Molina ◽  
Celia M. Santi
Keyword(s):  
Human Sperm ◽  
Membrane Depolarization ◽  
Concentration Increase ◽  
Epididymal Sperm ◽  
Soluble Adenylyl Cyclase ◽  
Mouse Sperm ◽  
Intracellular Ca ◽  
Sperm Membrane ◽  
Human And Mouse ◽  
Kinase A

To fertilize an egg, mammalian sperm must undergo capacitation in the female genital tract. A key contributor to capacitation is the calcium (Ca2+) channel CatSper, which is activated by membrane depolarization and intracellular alkalinization. In mouse epididymal sperm, membrane depolarization by exposure to high KCl triggers Ca2+ entry through CatSper only in alkaline conditions (pH 8.6) or after in vitro incubation with bicarbonate (HCO3–) and bovine serum albumin (capacitating conditions). However, in ejaculated human sperm, membrane depolarization triggers Ca2+ entry through CatSper in non-capacitating conditions and at lower pH (< pH 7.4) than is required in mouse sperm. Here, we aimed to determine the mechanism(s) by which CatSper is activated in mouse and human sperm. We exposed ejaculated mouse and human sperm to high KCl to depolarize the membrane and found that intracellular Ca2+ concentration increased at pH 7.4 in sperm from both species. Conversely, intracellular Ca2+ concentration did not increase under these conditions in mouse epididymal or human epididymal sperm. Furthermore, pre-incubation with HCO3– triggered an intracellular Ca2+ concentration increase in response to KCl in human epididymal sperm. Treatment with protein kinase A (PKA) inhibitors during exposure to HCO3– inhibited Ca2+ concentration increases in mouse epididymal sperm and in both mouse and human ejaculated sperm. Finally, we show that soluble adenylyl cyclase and increased intracellular pH are required for the intracellular Ca2+ concentration increase in both human and mouse sperm. In summary, our results suggest that a conserved mechanism of activation of CatSper channels is present in both human and mouse sperm. In this mechanism, HCO3– in semen activates the soluble adenylyl cyclase/protein kinase A pathway, which leads to increased intracellular pH and sensitizes CatSper channels to respond to membrane depolarization to allow Ca2+ influx. This indirect mechanism of CatSper sensitization might be an early event capacitation that occurs as soon as the sperm contact the semen.

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.

scholarly journals ZP2 peptide beads select human sperm in vitro, decoy mouse sperm in vivo, and provide reversible contraception

2016 ◽  
Vol 8 (336) ◽  
pp. 336ra60-336ra60 ◽  
Author(s):  
Matteo A. Avella ◽  
Boris A. Baibakov ◽  
Maria Jimenez-Movilla ◽  
Anna Burkart Sadusky ◽  
Jurrien Dean
Keyword(s):  
Human Sperm ◽  
Mouse Sperm

scholarly journals Soluble adenylyl cyclase inhibition prevents human sperm functions essential for fertilization

2021 ◽  
Author(s):  
Melanie Balbach ◽  
Lubna Ghanem ◽  
Thomas Rossetti ◽  
Navpreet Kaur ◽  
Carla Ritagliati ◽  
...  
Keyword(s):  
Adenylyl Cyclase ◽  
Ex Vivo ◽  
Beat Frequency ◽  
Human Sperm ◽  
Soluble Adenylyl Cyclase ◽  
Mouse Sperm ◽  
Acrosomal Exocytosis ◽  
Induced Changes ◽  
First Time ◽  
Sperm Functions

Abstract Soluble adenylyl cyclase (sAC: ADCY10) has been genetically confirmed to be essential for male fertility in mice and humans. In mice, ex vivo studies of dormant, caudal epididymal sperm demonstrated that sAC is required for initiating capacitation and activating motility. We now use an improved sAC inhibitor, TDI-10229, for a comprehensive analysis of sAC function in mouse and human sperm. In contrast to caudal epididymal mouse sperm, human sperm are collected post-ejaculation, after sAC activity has already been stimulated. In addition to preventing the capacitation-induced stimulation of sAC and protein kinase A activities, tyrosine phosphorylation, alkalinization, beat frequency, and acrosome reaction in dormant mouse sperm, sAC inhibitors interrupt each of these capacitation-induced changes in ejaculated human sperm. Furthermore, we show for the first time that sAC is required during acrosomal exocytosis in mouse and human sperm. These data define sAC inhibitors as candidates for non-hormonal, on-demand contraceptives suitable for delivery via intravaginal devices in women.

scholarly journals Localization of human sperm PMCA4b which is required for normal motility in mouse sperm

2010 ◽  
Vol 24 (S1) ◽  
Author(s):  
Katharine Shelly ◽  
Rolands Aravindan ◽  
Patricia Martin‐DeLeon
Keyword(s):  
Human Sperm ◽  
Mouse Sperm
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