Differing requirements for capacitation in vitro of mouse spermatozoa from two strains

Fraser, Lynn R.

doi:10.1530/jrf.0.0490083

Cited by 27 publications

(10 citation statements)

References 10 publications

(16 reference statements)

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“…It is known that when the high concentrations of sperm recovered from the cauda epididymis are cultured for more than 60 minutes (pre-culture), capacitation is induced spontaneously (Fraser, 1977). Therefore, we examined whether the prolonged pre-culture of sperm…”

Section: Research Articlementioning

confidence: 99%

Hyaluronan fragments generated by sperm-secreted hyaluronidase stimulate cytokine/chemokine production via the TLR2 and TLR4 pathway in cumulus cells of ovulated COCs, which may enhance fertilization

et al. 2008

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The toll-like receptor (TLR) system is expressed in cumulus cells of ovulated cumulus-oocyte complexes (COCs) and is activated by bacterial lipopolysaccharides (LPS). However, the endogenous ligand(s) for the TLRs and the physiological role(s) in ovulated COCs remain to be defined. Based on reports that hyaluronan fragments can activate TLR2 and TLR4 in macrophages, and that ovulated COCs are characterized by a hyaluronan-rich matrix, we cultured ovulated mouse COCs with purified hyaluronan fragments, treated them with purified hyaluronidase or exposed them to sperm as a physiologically relevant source of hyaluronidase. Hyaluronan fragments or hyaluronidase activated the NFkappaB pathway and induced Il6, Ccl4 and Ccl5 mRNA expression within 2 hours. Anti-TLR2 and anti-TLR4 neutralizing antibodies significantly suppressed hyaluronan fragment- and hyaluronidase-induced activation of the NFkappaB pathway and the expression of these genes. When ovulated COCs were cultured with sperm, the expression and secretion of cytokine/chemokine family members were induced in a time-dependent manner that could be blocked by TLR2/TLR4 antibodies or by a hyaluronan-blocking peptide (Pep-1). The chemokines secreted from TLR2/TLR4-stimulated COCs activated cognate chemokine receptors (CCRs) localized on sperm and induced sperm protein tyrosine phosphorylation, which was used as an index of capacitation. Significantly, in vitro fertilization of COC-enclosed oocytes was reduced by the TLR2/TLR4 neutralizing antibodies or by Pep-1. From these results, we propose that TLR2 and TLR4 present on cumulus cells were activated by the co-culture with sperm in a hyaluronan fragment-dependent manner, and that chemokines secreted from COCs induced sperm capacitation and enhanced fertilization, providing evidence for a regulatory loop between sperm and COCs during fertilization.

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Section: Research Articlementioning

confidence: 99%

Hyaluronan fragments generated by sperm-secreted hyaluronidase stimulate cytokine/chemokine production via the TLR2 and TLR4 pathway in cumulus cells of ovulated COCs, which may enhance fertilization

et al. 2008

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“…Thus, variations between strains in the rate or proportion of spermatozoa capacitating [Fraser and Drury, 19761 may be expected to produce different fertilization rates at different sperm concentrations. Apart from the reports by Fraser [1977] of differences in the capacitation period between spermatozoa from TO and F, (CBA x C57BL) mice, little information exists comparing the ability of strains to respond to fertilization conditions in vitro. Some data suggest the spermatozoa from different strains of mice may differ in their CAMP levels in the presence or absence of energy substrates [Nadyika and Hillman, 19801, in their calcium requirements [Parkening and Chang, 19761 and albumin requirements [Fraser and Drury, 19751. A significant decline in fertilization rates occurred at 8 x lo6 sperm/ml using F, spermatozoa, but with QS spermatozoa the fertilization values began to decrease at only 4 x lo6 sperm/ml (Table 1).…”

Section: Discuss10 Nmentioning

confidence: 99%

“…This report confirms earlier observations on the inhibitory capacity of concentrated sperm suspensions [Iwamatsu and Chang, 197 1 ;Oliphant and Brackett, 19731. Capacitation in some form must occur even in the presence of the inhibitor, since Fraser [1977] has reported that C57BL spermatozoa have a greater fertilizing capacity if preincubated for longer periods in a concentrated form. Spermatozoa maintained as a neat sperm suspension develop the ability to bind to the zona pellucida [Inoue and Wolf, 19751 even though fertilization does not proceed [Tsunoda and Chang, 19751.…”

Section: Discuss10 Nmentioning

confidence: 99%

Fertilization of cumulus‐free, zona‐intact mouse ova in vitro at high and low sperm concentrations

Stanger

Quinn

1982

Gamete Research

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The effect of varying the sperm concentration between 2 × 105 sperm/ml and 8 × 106 sperm/ml on fertilization of cumulus‐free, zona‐intact F1 (CBA × C57BL) mouse ova by QS and F1 (CBA × C57BL) mouse spermatozoa was studied. The spermatozoa from both strains of mice exhibited optimal fertilization rates at 2 × 106 sperm/ml. However, at sperm concentrations greater than 4 × 106 sperm/ml and less than 1 × 106 sperm/ml, fertilization rates were significantly reduced. F1 spermatozoa were more susceptible to dilution than QS spermatozoa. A significant interaction between strain and sperm concentration indicated that the two strains produced different fertilization rates at different sperm densities. Extracts of epididymal fluid, medium from capacitated spermatozoa, or ampulla fluid did not improve the fertilization rate at 2 × 105 sperm/ml, but retaining the cumulus oophorus did. The decrease in fertilization rate at 8 × 106 sperm/ml can in part be attributed to a nondialysable inhibitor from the neat sperm preparation that appeared to be of epididymal origin.

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“…The use of IVF for producing embryos for rederivation and/or embryo banking, even under some difficult circumstances can speed up and in some cases improve the process of cryopreservation. However, the success of the cryopreservation is influenced by the genetic background and the reproductive performance of the mice used (Choi et al 2000; Fraser 1977; Parkening and Chang 1976). A Number of differences exists between the mouse strains commonly used in the research field and in transgenic technology (e.g., B6; 129 × 1/SvJ, FVB/N and BALB/c).…”

Section: Discussionmentioning

confidence: 99%

“…Reproductive traits such as litter size, sperm production and hormone induced ovulation rate differ between mouse strains (Bouxsein et al 2005; Spearow and Barkley 1999; Silver 1995). Also the in vitro fertilization in mice varies depending on genotype and the conditions used for capacitation and fertilization (Sztein et al 2000; Choi et al 2000; Fraser 1977; Parkening and Chang 1976; Fraser and Drury 1976; Iwamatsu and Chang 1971). Further, inbred mouse strains have defined reproductive parameters which are so distinctive that they are considered to be a characteristic of the strain (Byers et al 2006; Silver 1995).…”

Section: Discussionmentioning

confidence: 99%

Fertility comparison between wild type and transgenic mice by in vitro fertilization

2009

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Transgenic mice are increasingly used as animal models for studies of gene function and regulation of mammalian genes. Although there has been continuous and remarkable progress in the development of transgenic technology over several decades, many aspects of the resulting transgenic model’s phenotype cannot be completely predicted. For example, it is well known that as a consequence of the random insertion of the injected DNA construct, several founder mice of the new line need to be analyzed for possible differences in phenotype secondary to different insertion sites. The Knock out technique for transgenic production disrupts a specific gene by insertion or homologous recombination creating a null expression or replacement of the gene with a marker to localize it expression. This modification could result in pleiotropic phenotype if the gene is also expressed in tissues other than the target organs. Although the future breeding performance of the newly created model is critical to many studies, it is rarely anticipated that the new integrations could modify the reproductive profile of the new transgenic line. To date, few studies have demonstrated the difference between the parent strain’s reproductive performance and the newly developed transgenic model. This study was designed to determine whether a genetic modification, knock out (KO) or transgenics, not anticipated to affect reproductive performance could affect the resulting reproductive profile of the newly developed transgenic mouse. More specifically, this study is designed to study the impact of the genetic modification on the ability of gametes to be fertilized in vitro. We analyzed the reproductive performance of mice with different background strains: FVB/N, C57BL/6 (129Sv/J × C57Bl/6)F1 and outbred CD1® and compared them to mice of the same strain carrying a transgene or KO which was not anticipated to affect fertility. In vitro Fertilization was used to analyze the fertility of the mice. Oocytes from superovulated females were inseminated with sperm of same background. Fertility rate was considered as the percentage of two cell embryos scored 24 h after insemination. The data collected from this study shows that the fertilization rate is affected (reduced to half fold) in some of the transgenic mice compared to the respective Wild Type (WT) mice. For the WT the average fertility rate ranged from 80% (C57BL/6), 90% (FVB/N), 45% (129Sv/J × C57Bl/6)F1 and 43% (CD1). For transgenic mice it was 52% (C57BL/6), 65% (FVB/N), 22% (129Sv/J × C57Bl/6)F1 and 25% (CD1).

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Differing requirements for capacitation in vitro of mouse spermatozoa from two strains

Abstract: Summary. Concentrated

Cited by 27 publications

References 10 publications

Hyaluronan fragments generated by sperm-secreted hyaluronidase stimulate cytokine/chemokine production via the TLR2 and TLR4 pathway in cumulus cells of ovulated COCs, which may enhance fertilization

Hyaluronan fragments generated by sperm-secreted hyaluronidase stimulate cytokine/chemokine production via the TLR2 and TLR4 pathway in cumulus cells of ovulated COCs, which may enhance fertilization

Fertilization of cumulus‐free, zona‐intact mouse ova in vitro at high and low sperm concentrations

Fertility comparison between wild type and transgenic mice by in vitro fertilization

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