Potassium dependence for sperm–egg fusion in mice

Boldt, Jeffrey; Casas, Adela T.; Whaley, Elizabeth; Creazzo, Tony L.; Lewis, Jill B.

doi:10.1002/jez.1402570215

Cited by 12 publications

(6 citation statements)

References 19 publications

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“…However, their experi-mental conditions differed from ours in that their cocultures consisted of one sperm with one or two eggs. On the other hand, Boldt et al (1991) obtained a result similar to ours, that is, a higher penetration rate of A23187-treated sperm than that of control sperm, although their treatment was performed at 10 pM A23187 in 2% BSA-containing medium.…”

Section: Discussionsupporting

confidence: 84%

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Tanphaichitr

Hansen

1994

Molecular Reproduction Devel

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A method to generate a population of motile, acrosome-reacted mouse sperm is described. Sperm retrieved from the cauda epididymis and vas deferens were first capacitated in a 3% bovine serum albumin (BSA) containing medium. Sperm were then resuspended in medium with low BSA content (0.01%) and treated with 30 nM of the calcium ionophore, A23187, which was added as a single dose of 30 nM for 15 min at 37 degrees C; or three sequential 10 nM doses over three 5 min intervals. Approximately 55-60% of the treated sperm population became acrosome reacted. The motility of the treated sperm sample was 40-65%, slightly lower than that of the control sperm, following addition of medium containing 3% BSA. This is in contrast to the < 10% motility observed for capacitated mouse sperm treated with 10 microM 23187, a concentration that had been used by other investigators to induce the acrosome reaction. The ultrastructure of the 30 nM A23187-induced acrosome-reacted sperm was similar to that of the acrosome-reacted sperm induced by solubilized zonae pellucidae. These motile, acrosome-reacted sperm were able to penetrate zona-free mouse eggs at a higher rate than the control sperm. Thus this method of treatment will be useful for further physiological experimentation with acrosome-reacted sperm.

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Section: Discussionsupporting

confidence: 84%

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Tanphaichitr

Hansen

1994

Molecular Reproduction Devel

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“…Sperm were obtained from retired breeder CFI males by removing the caudae epididymides and placing them into drops of culture media containing 2% BSA. The culture medium used was a modified Kreb's Ringer bicarbonate-buffered medium containing glucose and pyruvate as energy sources (Boldt et al, 1991). After 10 min to allow sperm to disperse, epididymal pieces were removed, and the sperm concentration was adjusted to 5 x 106/ml with 2% BSA media.…”

Section: In Vitro Fertilizationmentioning

confidence: 99%

“…After 10 min to allow sperm to disperse, epididymal pieces were removed, and the sperm concentration was adjusted to 5 x 106/ml with 2% BSA media. Sperm samples were incubated for 90 min to allow capacitation, then were treated with calcium ionophore A23187 (5-10 pM) for 30 min to induce acrosome reactions (Boldt et al, 1991). Inseminations were conducted in 200 pl drops of 2% BSA with 20-30 eggs and 1 x lo5 spermlml in the drops.…”

Section: In Vitro Fertilizationmentioning

confidence: 99%

Recovery of penetration ability in protease‐treated zona‐free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein

Kellom

Vick

Boldt

1992

Molecular Reproduction Devel

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Previous studies have demonstrated that protease treatment of zona-free mouse eggs impairs sperm-egg interaction (Boldt et al.: Biol Reprod 39:19-27, 1988) and causes modification of a 94 kD egg plasma membrane protein (Boldt et al., Gamete Res 23:91-101, 1989). In this report, the ability of eggs to recover penetration ability following protease treatment was examined. Zona-free mouse eggs were isolated and treated with either trypsin or chymotrypsin (1 mg/ml, 20 min), then cultured for 0, 3, or 6 hr before insemination. Eggs cultured for 3 or 6 hr displayed significantly higher penetration levels than eggs inseminated immediately after protease treatment, indicating a recovery of penetration ability during the 3 or 6 hr incubation period. The recovery of penetration ability was not blocked by inclusion of cyclohexamide (50 micrograms/ml) during the 3 or 6 hr culture period, indicating that protein synthesis was not required for recovery of fusion ability. Cell surface radiolabeling studies with 125I revealed that a 94 kD cell surface protein was lost immediately following trypsin or chymotrypsin treatment but was found on the egg surface after the 3 or 6 hr recovery period. Recovery of the 94 kD egg surface protein occurred in the presence of cyclohexamide, and metabolic radiolabeling studies with 35S-methionine confirmed that synthesis of a 94 kD protein was blocked by cyclohexamide. These results suggest that the recovery of penetration ability after protease treatment of zona-free eggs is due to recovery of the 94 kD cell surface protein, providing further evidence for the involvement of the 94 kD protein in sperm-egg interaction.

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“…and implantation of the developing embryo (Clemetson et al 1972;Boldt, Casas, Whaley, Creazzo & Lewis, 1991;Fraser, 1992Fraser, , 1995. Other important functions of endometrial epithelial cells include regulation of luminal pH and uterine fluid volume.…”

mentioning

confidence: 99%

Regulation of chloride secretion across porcine endometrial epithelial cells by prostaglandin E₂

Deachapunya

O’Grady

1998

The Journal of Physiology

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The objective of this study was to investigate the mechanism of PGE2 regulation of Cl− transport across glandular endometrial cells grown in primary culture. Most of the basal short circuit current (Isc) was inhibited by luminal addition of 5‐nitro‐2‐(3‐phenylpropylamino)benzoic acid (NPPB) or glibenclamide, suggesting the presence of a basally active Cl− conductance in the apical membrane. Basolateral addition of 10 μM PGE2 increased Isc by 41 ± 3 μA. A similar response was observed when cells were treated with 8‐(4‐chlorophenylthio) adenosine 3′,5′‐cyclic monophosphate (CPT‐cAMP). Pretreatment of monolayers with NPPB and glibenclamide blocked the PGE2 and cAMP‐mediated increase in Isc, suggesting that the effects of PGE2 and cAMP were dependent on the activity of an apical NPPB‐ and glibenclamide‐sensitive conductance. Addition of 50 nM antiPGE2 antibody to the basolateral bathing solution decreased basal Isc by 20 % and shifted the threshold response to exogenous PGE2. This result suggests autocrine regulation of electrogenic Cl− transport by PGE2. Experiments with amphotericin B‐permeabilized monolayers revealed that the apical PGE2‐activated, NPPB‐ and glibenclamide‐sensitive conductance was Cl− dependent and that the current‐voltage relationship and anion permeation properties (SCN−>Br− > Cl− > I−) were characteristic of the cystic fibrosis transmembrane conductance regulator (CFTR). Cultured porcine endometrial epithelial cells were specifically labelled with an antibody to a peptide sequence within the regulatory domain of CFTR. The effect of PGE2 was blocked by basolateral addition of bumetanide and furosemide at concentrations that are selective for inhibition of Na+‐K+‐2Cl−cotransport activity. The effect of bumetanide on Isc was Cl− dependent, suggesting a role for the bumetanide‐sensitive transport pathway in Cl− secretion. PGE2 and cAMP also activated an outwardly rectifying basolateral K+ channel which presumably sustains the driving force for electrogenic Cl− efflux across the apical membrane. The concentration‐conductance and concentration‐Isc response relationships for PGE2 showed that basolateral K+ permeability was rate limiting with respect to transepithelial anion secretion and that activation of a basolateral K+ channel by PGE2 was necessary to achieve maximum rates of Cl− secretion.

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Potassium dependence for sperm–egg fusion in mice

Cited by 12 publications

References 19 publications

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Recovery of penetration ability in protease‐treated zona‐free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein

Regulation of chloride secretion across porcine endometrial epithelial cells by prostaglandin E₂

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Potassium dependence for sperm–egg fusion in mice

Cited by 12 publications

References 19 publications

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187

Recovery of penetration ability in protease‐treated zona‐free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein

Regulation of chloride secretion across porcine endometrial epithelial cells by prostaglandin E2

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Regulation of chloride secretion across porcine endometrial epithelial cells by prostaglandin E₂