Assessment of Progesterone‐Induced Acrosome Reaction by Biotinylated Monoclonal Antibody Probes

Lee, Chi‐Yu Gregory; Khorasani, Afsaneh Motamed; Dorjee, Sonam

doi:10.1111/j.1600-0897.1998.tb00350.x

Cited by 3 publications

(4 citation statements)

References 28 publications

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“…Similar to a previous observation (Lee et al, 1998), the mean percent changes of acrosome-reacted sperm increased significantly in the presence of P 4 in 8 repeated experiments (Table 1). At a P 4 concentration of 10 g/ mL, the percent change of acrosome-reacted sperm increased significantly by 7.3% Ϯ 1.1% and 10.7% Ϯ 1.2% for sperm that were capacitated for 2 and 18 hours, respectively (P Ͻ .05 compared with ␤-cyclodextrin as the control, in which acrosome reaction increased by 1.3% Ϯ 0.5%; Table 1).…”

Section: Progesterone-induced Acrosome Reactionsupporting

confidence: 90%

“…Highly motile sperm were prepared for capacitation using a modified HTF medium at 37ЊC and 5% CO 2 and incubated for 2 to 18 hours. Progesterone (10 g/mL) was then added to the preparation for 30 minutes to induce an acrosome reaction as described by Lee et al (1998). Acrosome status was assessed using an FITC-PSA probe.…”

Section: Induction Of the Acrosome Reaction By Progesteronementioning

confidence: 99%

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Cholesterol Inhibitory Effects on Human Sperm—Induced Acrosome Reaction

KHORASANI,

CHEUNG,

LEE

2000

Journal of Andrology

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Progesterone (P4) is known to induce an acrosome reaction in mammalian sperm in vitro, whereas cholesterol is a major inhibitor of acrosome reaction. This study had three objectives: to study the in vitro effects of exogenous cholesterol on acrosome reactions in human sperm, to study the mechanism by which cholesterol affects P4‐induced acrosome reaction and those induced by dibutyryl cyclic adenosine monophosphate (db‐cAMP), and to study the status of the P4 surface receptor during capacitation and acrosome reaction and its relationship with cholesterol and different acrosome reaction inducers. Acrosome reaction was induced with exposure to 10 μg/mL of P4 for 30 minutes and 1 mM of db‐cAMP for 30 minutes in motile sperm either in the presence or absence of 0.1–1 μg/mL of cholesterol for 30 minutes. The effects of a 30‐minute exposure to 1 μg/mL of β‐sitosterol, a cholesterol plant analogue, as well as the effects of cholesterol on P4‐induced acrosome reactions were compared. Fluorescein isothiocyanate‐labeled albumin‐progesterone conjugate (P4‐FITC‐BSA) was used as the probe in order to quantify the percentage of sperm in which the P4 surface receptor was exposed. The results of this study indicate that cholesterol inhibited P4‐induced acrosome reactions when added to the sperm during capacitation (long incubation) and when it was added with P4 during the induction of acrosome reactions (short incubation). Similarly, acrosome reaction that was induced by db‐cAMP was also inhibited by cholesterol. Fifty percent of P4‐induced acrosome reaction was inhibited by a cholesterol concentration of 0.2 μg/mL Cholesterol's inhibition of induced acrosome reaction was independent of P4 concentration. β‐Sitosterol inhibited P4‐induced acrosome reaction in a dose‐dependent manner that was identical to that of cholesterol. We observed that increases in the P4 surface receptor exposure were time‐dependent and receptors migrated toward the equatorial segment during the first 2 hours of capacitation. We also found that db‐cAMP induced the appearance of the P4 surface receptor in the sperm plasma membrane and that cholesterol inhibited it. The results of this study suggest that cholesterol inhibits acrosome reaction in a noncompetitive manner by modifying the structure of the sperm plasma membrane, which prevents exposure of the P4 surface receptor for P4 binding.

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Section: Progesterone-induced Acrosome Reactionsupporting

confidence: 90%

Section: Induction Of the Acrosome Reaction By Progesteronementioning

confidence: 99%

Cholesterol Inhibitory Effects on Human Sperm—Induced Acrosome Reaction

KHORASANI,

CHEUNG,

LEE

2000

Journal of Andrology

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“…However, a recent report describes a very specific stimulation of progesterone binding to the physiologically important human sperm membrane progesterone receptor by digitonin (Ambhaikar & Puri 1998). The process of capacitation of human sperm is associated with a depletion of sperm membrane cholesterol, and the action of progesterone on the induction of the acrosome reaction can be antagonized by repletion of sperm membrane cholesterol (Cross 1996, Cross & Mahasrehti 1997, Lee et al 1998. We suggest that digitonin may complex with unesterified membrane sterol(s), 'chemically depleting' membrane cholesterol and unmasking latent membrane progesterone receptors.…”

Section: Effects Of Digitonin On Progesterone Bindingmentioning

confidence: 99%

Bovine ovarian non-genomic progesterone binding sites: presence in follicular and luteal cell membranes

Rae

Menzies²,

Bramley³

1998

Journal of Endocrinology

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We have shown recently that the bovine corpus luteum (CL) possesses specific luteal cell surface membrane binding sites for progesterone. We have now confirmed and extended these observations to compare the subcellular distribution of these binding sites in developing, mature and regressing CL. The median buoyant densities of luteal progesterone binding sites from early-, mid-and late-luteal phase CL were similar (though three of five density profiles for late-luteal phase CL showed association of steroid binding with a fraction with a lower density), and clearly resolved from nuclear, mitochondrial, lysosomal, peroxisomal, Golgi-endoplasmic reticulum-lysosomal and smooth endoplasmic reticulum markers. Specific binding of [3 H]progesterone overlapped with the distributions of 5 -nucleotidase and luteinizing hormone receptor (luteal cell surface membrane markers) in both control and digitonin-treated gradients at all stages of the luteal phase.Since steroidogenic 'large luteal' and 'small luteal' cells of the CL are derived from the granulosa cells (GC) and theca of the preovulatory follicle, we also investigated whether similar receptors were present in the follicle, and describe for the first time specific membrane binding sites for progesterone in purified GC and thecal membranes from healthy bovine follicles of different sizes. Specific binding increased linearly with GC and thecal membrane protein concentration; however, it was detectable only when digitonin was included in the binding incubation. Binding sites were specific for progesterone; unlabelled progesterone competed for [ 3 H]progesterone binding at low concentrations (IC 50 , 35 and 31 nmol/l) compared with testosterone (IC 50 , 905 and 870 nmol/l) and 4 -androstenedione (IC 50 , 1050 and 660 nmol/l) for GC and thecal receptors respectively. In contrast, oestradiol, oestrone, pregnenolone, cortisol, cholesterol, and a genomic progesterone receptor antagonist, RU486, competed poorly.Steroid binding was present in GC and thecal membranes of follicles of all sizes, but [ 3 H]progesterone binding to GC membranes decreased significantly with increasing follicle size (P<0·02), perhaps indicating developmental regulation of GC membrane non-genomic progesterone receptors in the preovulatory bovine follicle. We suggest that these membrane steroid receptors may be involved in the autocrine/paracrine regulation of follicular function by progesterone.

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“…Sperm were capacitated for 4 hours in IVF medium and induced to undergo the acrosome reaction with 5 µmol/L progesterone (Sigma) in IVF medium for 20 minutes as published. 15 A total of 25 zona-free oocytes (5 oocytes per experiment, experiments repeated 5 times) and acrosome-reacted sperm (1 × 10 6 sperm/mL) were incubated in IVF media droplets containing antiserum at 1:50 dilution for 2 hours in a humidified incubator at 38.5°C in 5% CO 2.…”

Section: Methodsmentioning

confidence: 99%

The Role of SPRASA in Female Fertility

et al. 2015

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Fertility is a complex process and infertility can have many causes. Sperm protein reactive with antisperm antibody (SPRASA)/sperm lysozyme-like protein 1 is a protein discovered as the target of autoantibodies in infertile men and previously thought to be expressed only in sperm. Using a bovine in vitro fertilization model, we have shown that SPRASA antiserum reduced sperm binding to zona-free oocytes and the development of embryos to morulae but did not affect the postfertilization cleavage rate to 2 cells or sperm motility. We demonstrated that SPRASA was expressed in ovarian follicles, corpora lutea, and oocytes by a combination of reverse transcription-polymerase chain reaction and immunohistochemistry. Female mice immunized with SPRASA had profound infertility following timed matings and those mice that did become pregnant had reduced fetal viability. The levels of antibodies reactive with SPRASA in 204 fertile and 202 infertile couples were elevated in 3 infertile but no fertile women. Together, these results indicate that SPRASA has a role in female fertility.

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Assessment of Progesterone‐Induced Acrosome Reaction by Biotinylated Monoclonal Antibody Probes

Cited by 3 publications

References 28 publications

Cholesterol Inhibitory Effects on Human Sperm—Induced Acrosome Reaction

Cholesterol Inhibitory Effects on Human Sperm—Induced Acrosome Reaction

Bovine ovarian non-genomic progesterone binding sites: presence in follicular and luteal cell membranes

The Role of SPRASA in Female Fertility

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