Techniques for Imaging Ca&lt;sup&gt;2+&lt;/sup&gt; Signaling in Human Sperm

Nash, Katherine; Lefièvre, Linda; Peralta-Arias, Ruben; Morris, Jennifer; Morales-Garcia, Aduen; Connolly, Tom; Costello, Sarah; Kirkman-Brown, Jackson; Publicover, Stephen J.

doi:10.3791/1996-v

Cited by 2 publications

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“…Sperm [Ca 2+ ] i changes were assessed using Fluo‐4 AM as described previously (Nash et al, 2010). The sperm were treated with 5 μM Fluo‐4 AM along with 0.05% Pluronic F‐127 (Molecular Probes) and subjected to a 30 min incubation at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Vaginal extracellular vesicles impair fertility in endometriosis by favoring Th17/Treg imbalance and inhibiting sperm activity

Zhang,

Xiong,

Jiang

et al. 2024

Journal Cellular Physiology

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Extracellular vesicles (EVs) play a key role in various diseases. However, their effect on endometriosis (EMs)‐associated infertility is poorly understood. We co‐cultured EVs from the female vaginal secretions with human sperm and also generated a mouse model of EMs by allogenic transplant to explore the effect of EVs on fertility. EVs from individuals with EMs‐associated infertility (E‐EVs) significantly inhibited the total motility (26.46% vs. 47.1%), progressive motility (18.78% vs. 41.06%), linear velocity (21.98 vs. 41.91 µm/s) and the acrosome reaction (AR) rate (5% vs. 22.3%) of human sperm in contrast to the control group (PBS). Furthermore, E‐EVs dose‐dependently decreased the intracellular Ca2+ ([Ca2+]i), a pivotal regulator of sperm function. Conversely, healthy women (H‐EVs) increased human sperm motion parameters, the AR rate, and sperm [Ca2+]i. Importantly, the mouse model of EMs confirmed that E‐EVs further decreased the conception rate and the mean number of embryo implantations (7.6 ± 3.06 vs. 4.5 ± 3.21) compared with the control mice by inducing the production of inflammatory cytokines leading to a Th17/Treg imbalance. H‐EVs could restore impaired fertility by restoring the Th17/Treg balance. We determined the impact of EVs derived from the female genital tract on human sperm function and studied the possible mechanisms by which it affects fertility. Our findings provide a novel rationale to ameliorate EMs‐associated infertility.

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Section: Methodsmentioning

confidence: 99%

Vaginal extracellular vesicles impair fertility in endometriosis by favoring Th17/Treg imbalance and inhibiting sperm activity

Zhang,

Xiong,

Jiang

et al. 2024

Journal Cellular Physiology

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“…Imaging was carried out as described ( Nash et al , 2010 ). Briefly, after adjusting sperm concentration to 1.5×10 6 /ml, the cell suspension was divided into aliquots of 200 μl and incubated with fluo4-AM (5 μM) for 30 min (36 °C, 5.5% CO 2 ).…”

Section: Methodsmentioning

confidence: 99%

SKF96365 modulates activity of CatSper channels in human sperm

Torrezan-Nitao

Brown

Lefièvre

et al. 2023

Molecular Human Reproduction

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Exposure of human sperm to progesterone (P4) activates cation channel of sperm (CatSper) channels, inducing an intracellular Ca2+ concentration ([Ca2+]i) transient followed by repetitive [Ca2+]i activity (oscillations), which are believed to be functionally important. We investigated the potential significance of store-operated Ca2+-entry in these oscillations using the inhibitor SKF96365 (30 µM; SKF). Following pre-treatment of human sperm with 3 µM P4, exposure to SKF doubled the proportion of oscillating cells (P = 0.00004). In non-pre-treated cells, SKF had an effect similar to P4, inducing a [Ca2+]i transient in > 80% of cells which was followed by oscillations in ≈50% of cells. The CatSper blocker RU1968 (11 µM) inhibited the SKF-induced [Ca2+]i increase and reversibly arrested [Ca2+]i oscillations. Using whole-cell patch clamp, we observed that SKF enhanced CatSper currents by 100% within 30 s, but amplitude then decayed to levels below control over the next minute. When cells were stimulated with P4, CatSper currents were stably increased (by 200%). Application of SKF then returned current amplitude to control level or less. When sperm were prepared in medium lacking bovine serum albumin (BSA), both P4 and SKF induced a [Ca2+]i transient in > 95% of cells but the ability of SKF to induce oscillations was greatly reduced (P = 0.0009). We conclude that SKF, similar to a range of small organic molecules, activates CatSper channels, but that a secondary blocking action also occurs, which was detected only during patch-clamp recording. The failure of SKF to induce oscillations when cells were prepared without BSA emphasises that the drug does not fully mimic the actions of P4.

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Techniques for Imaging Ca<sup>2+</sup> Signaling in Human Sperm

Cited by 2 publications

References 0 publications

Vaginal extracellular vesicles impair fertility in endometriosis by favoring Th17/Treg imbalance and inhibiting sperm activity

Vaginal extracellular vesicles impair fertility in endometriosis by favoring Th17/Treg imbalance and inhibiting sperm activity

SKF96365 modulates activity of CatSper channels in human sperm

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