The kinetics of the acrosome reaction of human spermatozoa and its correlation with in vitro fertilization

Takahashi, Kentaro; Wetzels, Alex M.M.; Goverde, H.J.M.; Bastiaans, Bart A.; Janssen, Hettie J. E. M.; Rolland, R.

doi:10.1016/s0015-0282(16)54976-0

Cited by 48 publications

(19 citation statements)

References 20 publications

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“…Multiple follicular development was induced by the administration of hMG (Humigon; Organon, Oss, the Netherlands) as described previously (9). A GnRH agonist (1 mg, Lucrin; Abbot, Amstelveen, the Netherlands) was administered SC from day 21 of the preceding cycle until the evening of hCG (Pregnyl; Organon) injection.…”

Section: P a Tien Tsmentioning

confidence: 99%

Glutathione S-transferases in human ovarian follicular fluid

Bisseling¹,

Knapen²,

Goverde³

et al. 1997

Fertility and Sterility

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Section: P a Tien Tsmentioning

confidence: 99%

Glutathione S-transferases in human ovarian follicular fluid

Bisseling¹,

Knapen²,

Goverde³

et al. 1997

Fertility and Sterility

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“…As exocytosis progresses, ligand bound-PNA diffuses out of the cell and the fluorescent signal is lost. The kinetics of AE have been previously shown to vary across individuals [22], therefore utilizing the time-sensitive lectin PNA on live cells is unsuitable for high-throughput analysis. Membrane fixation and permeabilization is necessary in order for PNA to bind to acrosomal ligands of intact cells [7, 21, 23], and thus, viability is not evaluated.…”

Section: Introductionmentioning

confidence: 99%

Glycopolymer induction of mouse sperm acrosomal exocytosis shows highly cooperative self-antagonism

Rodolis

Huang

Sampson

2016

Biochemical and Biophysical Research Communications

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Identifying inducers of sperm acrosomal exocytosis (AE) to understand sperm functionality is important for both mechanistic and clinical studies in mammalian fertilization. Epifluorescence microscopy methods, while reproducible, are laborious and incompatible for high throughput screening. Flow cytometry methods are ideal for quantitative measurements on large numbers of samples, yet typically rely on the use of lectins that can interfere with physiologic AE-inducers. Here, we present an optimized triple stain flow cytometric method that is suitable for high-throughput screening of AE activation by glycopolymers. SYTO-17 and propidium iodide (PI) were used to differentiate cells based on their membrane integrity or viability, and membrane impermeable soybean trypsin inhibitor (SBTI) was used to monitor acrosome exocytosis. The SBTI/PI/SYTO-17 combination provides a positive screen for viability and AE of live sperm cells with minimal noise or false positives. A scattering gate enables the use of samples that may be contaminated with non-cellular aggregates, e.g., cryopreservation agents. This assay format enabled detailed analysis of glycopolymer dose response curves. We found that fucose polymer has a narrow effective dose range (EC50 = 1.6 µM; IC50 = 13.5 µM); whereas mannose polymer and β-N-acetylglucosamine polymer have broader effective dose ranges (EC50 = 1.2 µM and 3.4 µM, respectively). These results highlight the importance of testing inducers over a large concentration range in small increments for accurate comparison.

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“…The first one did not react spontaneously with anti-CD46 antibodies (⌬⌿ m low , CD46 Ϫ spermatozoa) as ⌬⌿ m high spermatozoa, and the second one was characterized by binding of anti-CD46 antibodies (⌬⌿ m low , CD46 ϩ spermatozoa). Sperm with spontaneous AR lose their acrosomal membrane and thus are unable to bind to zona pellucida for oocyte fertilization (14). Only a minor population of spermatozoa (always Ͻ5%) were both ⌬⌿ m high and CD46 ϩ , indicating that spermatozoa with high ⌬⌿ m presented an intact acromome spontaneously (were CD46 Ϫ ; Fig.…”

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confidence: 97%