Increased disruption of sperm plasma membrane at sperm immobilization promotes dissociation of perinuclear theca from sperm chromatin after intracytoplasmic sperm injection in pigs

Katayama, Masahiro; Šutovský, Peter; Yang, Boh S; Cantley, T.C.; Rieke, August; Farwell, Randy; Oko, Richard; Day, Billy N.

doi:10.1530/rep.1.0680

Cited by 29 publications

(25 citation statements)

References 36 publications

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“…4). Interestingly, despite the presence of PLCz in both the head and the tail, injection of whole sperm induced normal fertilization in only 40% of the oocytes (Table 1) and similar results have been published by others (Katayama et al 2005). In contrast, following IVF, nearly 100% of sperm-penetrated oocytes display activation events resulting in PN formation (Kikuchi et al 1999).…”

Section: Discussionsupporting

confidence: 86%

Pre-treatment of sperm reduces success of ICSI in the pig

Nakai¹,

Ito²,

Sato³

et al. 2011

Reproduction

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In pigs, although ICSI is a feasible fertilization technique, its efficiency is low. In general, injected pig sperm are insufficient to induce oocyte activation and embryonic development. Pretreatments for disrupting sperm membranes have been applied to improve the fertility of ICSI oocytes; however, we hypothesize that such pretreatment(s) may reduce the ability of the sperm to induce oocyte activation. We first evaluated the effects of sperm pretreatments (sonication (SO) to isolate the sperm heads from the tails, Triton X-100 (TX), and three cycles of repeated freezing/thawing (3!-FT) for disrupting sperm membranes) on the rate of pronucleus (PN) formation after ICSI. We found that oocytes injected with control (whole) sperm had higher rates of PN formation than those obtained after subjecting the sperm to SO, TX, and 3!-FT. The amounts of phospholipase Cz (PLCz), which is thought to be the oocyte-activating factor in mammalian sperm, in sperm treated by each method was significantly lower than that in whole untreated sperm. Furthermore, using immunofluorescence, it was found that in pig sperm, PLCz was localized to both the post-acrosomal region and the tail area. Thus we demonstrated for the first time that sperm pretreatment leads to a reduction of oocyte-activating capacity. Our data also show that in addition to its expected localization to the sperm head, PLCz is also localized in the tail of pig sperm, thus raising the possibility that injection of whole sperm may be required to attain successful activation in pigs.

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Section: Discussionsupporting

confidence: 86%

Pre-treatment of sperm reduces success of ICSI in the pig

Nakai¹,

Ito²,

Sato³

et al. 2011

Reproduction

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“…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 after ICSI using plasma membrane-removed porcine spermatozoa were reported (15,16). Here, we report that removal of both the plasma membrane and acrosome from mouse spermatozoa before ICSI not only accelerates the onset of oocyte activation but also results in improved embryonic development.…”

mentioning

confidence: 79%

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.

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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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“…taurine and hypotaurine; Petters and Wells 1993)). Also, the benefits induced by CYS reported by Katayama et al (2007) may have been conditioned by combining CYS with the use of piezo pulses for sperm-immobilization to disrupt the sperm plasma membrane, which it is known to improve the fertilization rates and the decondensation of the sperm chromatin after ICSI (Katayama et al 2005). It should be highlighted that in our study the CYS supplement was tested alone, without any other sperm membrane disruptor than the sperm mid-piece rubbing step during ICSI (GarciaMengual et al 2011).…”

Section: Discussionmentioning

confidence: 99%

“…However, and contrarily to what happens after IVF, MPN formation of porcine oocytes in vitro matured with CYS is not improved after ICSI. The persistence of an intact sperm acrosome, plasma membrane and perinuclear theca (Katayama et al 2002a(Katayama et al , 2005, which are normally removed during sperm penetration in IVF and natural fertilization (Yanagimachi 1994), may be some of the factors causing an incorrect MPN formation in ICSI.…”

Section: Introductionmentioning

confidence: 99%

Male pronucleus formation after ICSI: effect of oocyte cysteine or sperm Triton X-100 treatments

García-Mengual¹,

Silvestre²,

Salvador³

et al. 2015

CZECH J ANIM SCI

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ABSTRACT:In pigs, intracytoplasmic sperm injection (ICSI) efficiency is still poor. The inadequate decondensation of the sperm chromatin, its transformation into the male pronucleus (MPN) together with the subsequent inability to activate the oocyte, seem to be the main causes of the low ICSI efficiency. In order to improve the MPN formation we took two different approaches. On the one hand, the in vitro culture (IVC) medium post-ICSI was supplemented with 1.71mM cysteine (CYS). Alternatively, the sperm membrane was digested with Triton X-100 (TX) before ICSI, to improve the exposure of the sperm chromatin to the oocyte cytoplasm. After 6 h post-ICSI, the activation rate was significantly higher in TX group (70.0%) compared with CYS and control groups (42.2% and 48.9%, respectively; P < 0.05). However, no significant differences between the three groups were observed in terms of the number of pronuclei, 2PN (oocytes with 2 pronuclei and no visible sperm), and 1PN + sperm (oocytes with 1 pronucleus and one sperm head). At 22 h post-ICSI, the activation rates were similar in TX, CYS, and control groups (73.1, 78.9, and 75.7%, respectively). In addition, we did not observe significant differences between TX, CYS, and control groups for the number of pronuclei, 2PN (52.6, 56.7, and 50%, respectively) or 1PN + sperm (21.1, 33.3, and 32.1%, respectively). While no cleavage was observed in the CYS group, no significant differences in the cleavage rate were observed between control (21.3%) and TX (10.5%) groups. In summary, and under our conditions, neither CYS supplement, nor sperm TX pre-treatment were able to improve MPN formation at 6 and 22 h post-ICSI. However, the sperm TX pre-treatment improved oocyte activation at 6 h post-ICSI, although 22 h post-ICSI such a beneficial effect did not persist.Keywords: porcine; assisted reproductive technology; pronuclear formation List of abbreviations: ICSI = intracytoplasmic sperm injection, PN = pronucleus, MPN = male pronucleus, IVC = in vitro culture, CYS = cysteine, TX = Triton X-100, IVP = in vitro production, IVM = in vitro maturation, IVF = in vitro fertilization, EGF = epidermal growth factor, GSH = glutathione, COC = cumulus oocyte complex, PZM-3 = porcine zygote medium-3, MM199 = maturation medium, HM199 = handling medium 242

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Increased disruption of sperm plasma membrane at sperm immobilization promotes dissociation of perinuclear theca from sperm chromatin after intracytoplasmic sperm injection in pigs

Cited by 29 publications

References 36 publications

Pre-treatment of sperm reduces success of ICSI in the pig

Pre-treatment of sperm reduces success of ICSI in the pig

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

Male pronucleus formation after ICSI: effect of oocyte cysteine or sperm Triton X-100 treatments

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