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

Tanphaichitr, Nongnuj; Hansen, C.

doi:10.1002/mrd.1080370312

Cited by 19 publications

(14 citation statements)

References 42 publications

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“…Conversely, sperm exposed to a ten-fold increase in cauda epididymal calcium concentration in mice mutant in TRPV6 (transient receptor potential vanilloid 6) that is a calcium ion selective channel, display increased intracellular calcium and have a markedly reduced motility and fertilization capacity [33]. In addition, sperm induced with 10 µM of the calcium ionophore A23187 will undergo the AR, but will become immotile [34].…”

Section: Resultsmentioning

confidence: 99%

Partial Deletion of Chromosome 8 β-defensin Cluster Confers Sperm Dysfunction and Infertility in Male Mice

et al. 2013

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β-defensin peptides are a family of antimicrobial peptides present at mucosal surfaces, with the main site of expression under normal conditions in the male reproductive tract. Although they kill microbes in vitro and interact with immune cells, the precise role of these genes in vivo remains uncertain. We show here that homozygous deletion of a cluster of nine β-defensin genes (DefbΔ9) in the mouse results in male sterility. The sperm derived from the mutants have reduced motility and increased fragility. Epididymal sperm isolated from the cauda should require capacitation to induce the acrosome reaction but sperm from the mutants demonstrate precocious capacitation and increased spontaneous acrosome reaction compared to wild-types but have reduced ability to bind the zona pellucida of oocytes. Ultrastructural examination reveals a defect in microtubule structure of the axoneme with increased disintegration in mutant derived sperm present in the epididymis cauda region, but not in caput region or testes. Consistent with premature acrosome reaction, sperm from mutant animals have significantly increased intracellular calcium content. Thus we demonstrate in vivo that β-defensins are essential for successful sperm maturation, and their disruption leads to alteration in intracellular calcium, inappropriate spontaneous acrosome reaction and profound male infertility.

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Section: Resultsmentioning

confidence: 99%

Partial Deletion of Chromosome 8 β-defensin Cluster Confers Sperm Dysfunction and Infertility in Male Mice

et al. 2013

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“…However, excessive intracellular calcium may induce microtubule defects and premature AR39. Such sperm will lose motility50 and the ability to firmly bind to the zona pellucida39 despite being capable of penetrating the egg5152. It is currently unclear whether such an excessive increase in intracellular calcium level occurs in the absence of PRM1, and further studies will be required to address this question.…”

Section: Discussionmentioning

confidence: 99%

Viable offspring obtained from Prm1-deficient sperm in mice

Takeda

Yoshinaga

Furushima

et al. 2016

Sci Rep

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Protamines are expressed in the spermatid nucleus and allow denser packaging of DNA compared with histones. Disruption of the coding sequence of one allele of either protamine 1 (Prm1) or Prm2 results in failure to produce offspring, although sperm with disrupted Prm1 or Prm2 alleles are produced. Here, we produced Prm1-deficient female chimeric mice carrying Prm1-deficient oocytes. These mice successfully produced Prm1+/− male mice. Healthy Prm1+/− offspring were then produced by transferring blastocysts obtained via in vitro fertilization using zona-free oocytes and sperm from Prm1+/− mice. This result suggests that sperm lacking Prm1 can generate offspring despite being abnormally shaped and having destabilised DNA, decondensed chromatin and a reduction in mitochondrial membrane potential. Nevertheless, these mice showed little derangement of expression profiles.

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“…According to Ajduk et al (28), pretreatment of mouse spermatozoa with 20 M Ca 2ϩ ionophore synchronizes pronuclear development of mouse oocytes after ICSI. Because such high concentrations of Ca 2ϩ ionophore immobilizes (''kills'') most spermatozoa (29), the synchronization of pronuclear development Ajduk et al (28) observed is likely because of extensive damage or loss of sperm plasma membranes by influx of excessive Ca 2ϩ . Mouse ICSI using LL-treated spermatozoa resulted in a high percentage of normal live offspring (Table 2).…”

Section: Discussionmentioning

confidence: 99%

Simultaneous removal of sperm plasma membrane and acrosome before intracytoplasmic sperm injection improves oocyte activation/embryonic development

Morozumi

Shikano

Miyazaki

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

100

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Direct injection of a single spermatozoon into an oocyte (ICSI) can produce apparently normal offspring. Although the production of normal offspring by ICSI has been successful in mice and humans, it has been less successful in many other species. The reason for this is not clear, but could be, in part, due to inconsistent activation of oocytes because of delayed disintegration of sperm plasma membrane within oocytes and incorporation of the acrosome containing a spectrum of hydrolyzing enzymes. In the mouse, the removal of sperm plasma membrane and acrosome was not a prerequisite to produce offspring by ICSI, but it resulted in earlier onset of oocyte activation and better embryonic development. The best result was obtained when spermatozoa were demembranated individually immediately before ICSI by using lysolecithin, a hydrolysis product of membrane phospholipids.mouse ͉ human ͉ Ca 2ϩ oscillations ͉ fertilization ͉ lysolecithin D irect injection of a single spermatozoon into an oocyte, commonly called intracytoplasmic sperm injection (ICSI), can produce apparently normal offspring even though it bypasses a number of biological processes necessary for normal fertilization. As long as the sperm nucleus has intact genetic integrity, ICSI can produce healthy offspring regardless of concentrations, morphology, and motility of spermatozoa (1, 2). Only one genomically normal spermatozoon is needed to fertilize one oocyte. A salient difference between natural and ICSI fertilization is that, in the latter, the sperm plasma membrane as well as acrosome (which contains a spectrum of powerful hydrolyzing enzymes) are introduced into an oocyte. For species with small acrosomes, injection of the acrosome into an oocyte apparently does not produce serious problems, but for species like the hamster, with very large acrosomes, injection inevitably results in death of the oocyte (3). We had demonstrated that the contents of the acrosome are potentially harmful to oocytes (4). A notable difference between normal and ICSI fertilization is that repetitive transient increases in intracellular Ca 2ϩ concentration of the oocyte (Ca 2ϩ oscillations), the pivotal signal for oocyte activation (5-8), begins much more slowly in ICSI oocytes than in normally fertilized oocytes. In the mouse, for instance, Ca 2ϩ oscillations begin 1-3 min after plasma membrane fusion between a fertilizing spermatozoon and an oocyte (9), whereas oscillation begins 15-30 min after ICSI (10, 11). In human oocytes, Ca 2ϩ oscillations start 4-12 h after ICSI, when a plasma membrane-intact spermatozoon is injected after immobilization by touching the terminal part of the tail (12). Ca 2ϩ oscillations begin faster (14 Ϯ 6 min) when a spermatozoon is immobilized by applying several piezo pulses to the proximal one-third of the sperm tail before injection (13). Kasai et al. (14) reported that oocyte activation, assessed by the completion of meiosis, occurred earlier when spermatozoa were freed from the plasma membrane before ICSI. Increased fertilization rates aft...

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

Cited by 19 publications

References 42 publications

Partial Deletion of Chromosome 8 β-defensin Cluster Confers Sperm Dysfunction and Infertility in Male Mice

Partial Deletion of Chromosome 8 β-defensin Cluster Confers Sperm Dysfunction and Infertility in Male Mice

Viable offspring obtained from Prm1-deficient sperm in mice

Simultaneous removal of sperm plasma membrane and acrosome before intracytoplasmic sperm injection improves oocyte activation/embryonic development

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