Calcium requirements for human sperm function in vitro

Marı́n-Briggiler, Clara I.; González-Echeverría, Fernanda; Buffone, Mariano G.; Calamera, J.C.; Tezón, Jorge G.; Vazquez‐Levin, Mónica H.

doi:10.1016/s0015-0282(03)00267-x

Cited by 35 publications

(31 citation statements)

References 41 publications

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“…In Ca 2+ -containing medium (2.5 mM), the percentage of hyperactivated cells was 19±3%. As reported (Marin-Briggiler et al, 2003), the development of this type of motility was significantly inhibited in -Ca 2+ medium (3±1%). Although incubation with KN62 reduced the percentage of cells displaying hyperactivation to 10±2%, a similar reduction was observed in the presence of KN04 (8±1%), suggesting a non-specific effect of these compounds on hyperactivation.…”

Section: Cam Kinase Inhibitors Decrease Human Sperm Motilitysupporting

confidence: 74%

“…Our results show that when human sperm are incubated in medium not supplemented with CaCl 2 , progressive motility is significantly reduced, and that 0.22 mM of this cation is sufficient to maintain flagellar motion. Similar Ca 2+ concentrations are required for the occurrence of some capacitation-related events such as the development of hyperactivated motility (Marin-Briggiler et al, 2003).…”

Section: Discussionmentioning

confidence: 94%

“…In human sperm, as well as in mouse sperm under certain conditions, the presence of Ca 2+ in the incubation medium decreases protein tyrosine phosphorylation levels (Carrera et al, 1996;Luconi et al, 1996;Marin-Briggiler et al, 2003;Baker et al, 2004). CaM (Carrera et al, 1996).…”

Section: +mentioning

confidence: 99%

“…Because previous works in sperm from different species (Marin-Briggiler et al, 2003;Marquez and Suarez, 2004) have shown that Ca 2+ is required for the development of hyperactivated motility, the possibility that CaM kinases are involved in the regulation of hyperactivation was also analysed. Hyperactivation parameters were investigated as described in human sperm incubated for 4 hours under capacitating conditions, in media supplemented or not with Ca 2+ and in the presence or in the absence of 60 µM KN62 or its inactive analogue KN04.…”

Section: Cam Kinase Inhibitors Decrease Human Sperm Motilitymentioning

confidence: 99%

“…In addition to the role of phosphorylation signalling in sperm motility, studies with a variety of species have shown that Ca 2+ ions play a key role in the development and maintenance of sperm progressive motion and hyperactivation (Lindemann et al, 1991;Serres et al, 1991;Marin-Briggiler et al, 1999;Marin-Briggiler et al, 2003;Marquez and Suarez, 2004). Using demembranated sperm models, some of these investigations have revealed that Ca 2+ levels modulate the waveform of the sperm tail (Ho et al, 2002;Lindemann and Goltz, 1988).…”

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confidence: 99%

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Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Marı́n-Briggiler

Jha

Chertihin

et al. 2005

Journal of Cell Science

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The mechanisms involved in the regulation of mammalian sperm motility are not well understood. Calcium ions (Ca2+) have been suggested to play a key role in the maintenance of motility; nevertheless, how Ca2+ modulates this process has not yet been completely characterized. Ca2+ can bind to calmodulin and this complex regulates the activity of multiple enzymes, including Ca2+/calmodulin-dependent protein kinases (CaM kinases). Results from this study confirmed that the presence of Ca2+ in the incubation medium is essential for maintaining human sperm motility. The involvement of CaM kinases in Ca2+ regulation of human sperm motility was evaluated using specific inhibitors (KN62 and KN93) or their inactive analogues (KN04 and KN92 respectively). Sperm incubation in the presence of KN62 or KN93 led to a progressive decrease in the percentage of motile cells; in particular, incubation with KN62 also reduced sperm motility parameters. These inhibitors did not alter sperm viability, protein tyrosine phosphorylation or the follicular fluid-induced acrosome reaction; however, KN62 decreased the total amount of ATP in human sperm. Immunological studies showed that Ca2+/calmodulin-dependent protein kinase IV (CaMKIV) is present and localizes to the human sperm flagellum. Moreover, CaMKIV activity increases during capacitation and is inhibited in the presence of KN62. This report is the first to demonstrate the presence of CaMKIV in mammalian sperm and suggests the involvement of this kinase in the regulation of human sperm motility.

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Section: Cam Kinase Inhibitors Decrease Human Sperm Motilitysupporting

confidence: 74%

Section: Discussionmentioning

confidence: 94%

Section: +mentioning

confidence: 99%

Section: Cam Kinase Inhibitors Decrease Human Sperm Motilitymentioning

confidence: 99%

mentioning

confidence: 99%

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Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Marı́n-Briggiler

Jha

Chertihin

et al. 2005

Journal of Cell Science

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Role of the Na⁺/Ca²⁺ exchanger in calcium homeostasis and human sperm motility regulation

Krasznai

Morisawa²,

Bazsáné

et al. 2005

Cell Motil. Cytoskeleton

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A number of cell functions, such as flagellar beating, swimming velocity, acrosome reaction, etc., are triggered by a Ca2+ influx across the cell membrane. For appropriate physiological functions, the motile human sperm maintains the intracellular free calcium concentration ([Ca2+]i) at a submicromolar level. The objective of this study was to determine the role of the Na+/Ca2+ exchanger (NCX) in the maintenance of [Ca2+]i in human spermatozoa. Spermatozoa maintained in extracellular medium containing>or=1 microM Ca2+ exhibited motility similar to that of the control. In addition to several calcium transport mechanisms described earlier, we provide evidence that the NCX plays a crucial role in the maintenance of [Ca2+]i. Three chemically unrelated inhibitors of the NCX (bepridil, DCB (3',4'-dichlorobenzamil hydrochloride), and KB-R7943) all blocked human sperm motility in a dose and incubation time dependent manner. The IC50 values for bepridil, DCB, and KB-R7943 were 16.2, 9.8, and 5.3 microM, respectively. The treatment with the above-mentioned blockers resulted in an elevated [Ca2+]i and a decreased [Na+]i. The store-operated calcium channel (SOCC) inhibitor SKF 96365 also blocked the sperm motility (IC50=2.44 microM). The presence of the NCX antigen in the human spermatozoa was proven by flow cytometry, confocal laser scanning microscopy, and immunoblotting techniques. Calcium homeostasis of human spermatozoa is maintained by several transport proteins among which the SOCC and the NCX may play a major role.

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Only a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation

Luque

Dalotto-Moreno

Martín‐Hidalgo

et al. 2018

Journal Cellular Physiology

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Mammalian sperm must undergo a functionally defined process called capacitation to be able to fertilize oocytes. They become capacitated in vivo by interacting with the female reproductive tract or in vitro in a defined capacitation medium that contains bovine serum albumin, calcium (Ca ), and bicarbonate (HCO ). In this work, sperm were double stained with propidium iodide and the Ca dye Fluo-4 AM and analyzed by flow cytometry to determine changes in intracellular Ca concentration ([Ca ] ) in individual live sperm. An increase in [Ca ] was observed in a subpopulation of capacitated live sperm when compared with noncapacitated ones. Sperm exposed to the capacitating medium displayed a rapid increase in [Ca ] within 1 min of incubation, which remained sustained for 90 min. These rise in [Ca ] after 90 min of incubation in the capacitating medium was evidenced by an increase in the normalized median fluorescence intensity. This increase was dependent on the presence of extracellular Ca and, at least in part, reflected the contribution of a new subpopulation of sperm with higher [Ca ] . In addition, it was determined that the capacitation-associated [Ca ] increase was dependent of CatSper channels, as sperm derived from CatSper knockout (CatSper KO) or incubated in the presence of CatSper inhibitors failed to increase [Ca ] . Surprisingly, a minimum increase in [Ca ] was also observed in CatSper KO sperm suggesting the existence of other Ca transport systems. Altogether, these results indicate that a subpopulation of sperm increases [Ca ] very rapidly during capacitation mainly due to a CatSper-mediated influx of extracellular Ca .

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Calcium requirements for human sperm function in vitro

Cited by 35 publications

References 41 publications

Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Role of the Na⁺/Ca²⁺ exchanger in calcium homeostasis and human sperm motility regulation

Only a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation

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Calcium requirements for human sperm function in vitro

Cited by 35 publications

References 41 publications

Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation

Role of the Na+/Ca2+ exchanger in calcium homeostasis and human sperm motility regulation

Only a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation

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Role of the Na⁺/Ca²⁺ exchanger in calcium homeostasis and human sperm motility regulation