Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm

Kurokawa, Manabu; Sato, Kenichi; Wu, Hua; He, Chuan; Malcuit, Christopher; Black, Samuel J.; Fukami, Kiyoko; Fissore, Rafael A.

doi:10.1016/j.ydbio.2005.06.029

Cited by 94 publications

(117 citation statements)

References 63 publications

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“…In the present study, the concentration of porcine PLCz cRNA injected into porcine oocytes is the same as that of mouse PLCz injected into mouse oocytes. Recently, Kurokawa et al (2005) reported that a single porcine sperm contains upwards of 350 fg of porcine PLCz protein. It is necessary in the future study to determine the minimum concentration of porcine PLCz protein that can trigger oocyte activation and induce embryonic development.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase Cζ

Yoneda

Kashima²,

Yoshida³

et al. 2006

Reproduction

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The molecular mechanism by which sperm triggers Ca 2C oscillation, oocyte activation, and early embryonic development has not been clarified. Recently, oocyte activation has been shown to be induced by sperm-specific phospholipase Cz (PLCz). The ability of PLCz to induce oocyte activation is highly conserved across vertebrates. In the present study, porcine PLCz cDNA was identified and the nucleotide sequence was determined. The expression pattern of porcine PLCz mRNA during the period of postnatal testicular development was shown to be similar to that of mouse PLCz. PLCz mRNA expression in the pig and mouse was detected only in the testes when the elongated spermatids had differentiated, and was detected from day 96 after birth in the pig. Histological examination of porcine testis during the period of postnatal development revealed the presence of spermatozoa from day 110 after birth. These findings suggest that the synthesis of PLCz mRNA starts when spermiogenesis is initiated. Microinjection of porcine PLCz complementary RNA into porcine oocytes demonstrated that porcine PLCz has the ability to trigger repetitive Ca 2C transients in porcine oocytes similar to that observed during fertilization. It was also found that porcine PLCz cRNA has the potential to induce oocyte activation and initiate embryonic development up to the blastocyst stage.

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Section: Discussionmentioning

confidence: 99%

Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase Cζ

Yoneda

Kashima²,

Yoshida³

et al. 2006

Reproduction

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“…Porcine PLCz (pPLCz) was detected using anti-PLCz rabbit serum generated against a 19-mer sequence (MENKWFLSMVRDDFKGGKI) at the N-terminus of pPLCz (accession no. BAC78817) as described previously (Kurokawa et al 2005). After denaturation by boiling at 99.5 8C for 3 min, samples were separated by SDS-PAGE on 12.5% polyacrylamide gel and then transferred onto a PVDF membrane (GE Healthcare).…”

Section: Western Blottingmentioning

confidence: 99%

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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“…Evidence suggests that the factor is a male-specific PLC, Zeta1 (PLCZ1), which was first discovered following a screen for expression of sequence tags related to PLCs in testis [7]. It was subsequently shown that expression of cRNAs encoding for PLCZ1 in eggs initiated oscillations reminiscent of those induced by normal fertilization [7,8]. Additional studies demonstrated expression of PLCZ1 in the testes of other mammals and PLCZ1 cRNAs were able to induce oscillations in eggs of homologous and heterologous species including human PLCZ1 into human eggs [7,9,10].…”

Section: Introductionmentioning

confidence: 99%

“…Additional studies demonstrated expression of PLCZ1 in the testes of other mammals and PLCZ1 cRNAs were able to induce oscillations in eggs of homologous and heterologous species including human PLCZ1 into human eggs [7,9,10]. Further, injection of sperm extracts depleted of PLCZ1 failed to initiate oscillations [7,8,11]. Despite this evidence, demonstration of the unique and primary role of PLCZ1 on mammalian fertilization needs confirmation from genetic models where the gene is purposely eliminated.…”

Section: Introductionmentioning

confidence: 99%

Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters

Lee

Arny

Grow

et al. 2014

J Assist Reprod Genet

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Purpose This study was conducted to determine if expression of the testis-specific phospholipase C Zeta1 (PLCZ1) correlated with low success or fertilization failure after ICSI in patients with normal parameters after standard semen analysis (SA). Methods Couples <43 years with one or two failed or low fertilization ICSI cycles. Standard Semen Analysis (SA) was performed to determine sperm parameters in male partners, whereas females were evaluated for antral follicle counts (AFC), day 3 FSH levels and peak Estradiol (E2) levels. The presence of PLCZ1 in sperm was ascertained using Western blotting and Immunofluorescence (IF) analysis. The ability of sperm to initiate changes in the intracellular concentrations of free calcium ([Ca 2+ ] i ), which is characteristic of mammalian sperm, was performed after injection of human sperm into mouse eggs loaded with the Ca 2+ sensitive dye fura-2 AM. Results Male partners of couples with failed or low success ICSI fertilization but with normal SA parameters showed low expression levels of PLCZ1 as determined by western blotting and reduced fluorescent signal during IF studies. In addition, fewer of these males' sperm showed PLCZ1 expression and were able to initiate robust [Ca 2+ ] i oscillations upon injection into eggs. Conclusion Our data suggest that in patients with normal SA parameters but with repeated low fertilization or outright failed fertilization results after ICSI, abnormal PLCZ1 function should be considered as the underlying mechanism responsible for the failure of fertilization.

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Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm

Cited by 94 publications

References 63 publications

Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase Cζ

Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase Cζ

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

Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters

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