Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection

Purpose The purpose of this study is to describe impaired oocyte fertilization from phospholipase C-zeta (PLC-ζ) deficiency in normal-appearing sperm that was successfully treated using calcium (Ca 2+ ) ionophore with intracytoplasmic sperm injection (ICSI) of oocytes matured in vitro. Methods An infertile couple undergoing in vitro fertilization (IVF) experienced failed oocyte fertilization following ICSI with normal-appearing sperm. A semen sample collected from the patient was used to assess the expression of sperm PLC-ζ protein by Western blot analysis and immunofluorescence and PLC-ζ bioactivity by an in vitro model of Ca 2+ release. A second IVF cycle was performed using Ca 2+ ionophore with ICSI to enhance Ca 2+ -induced oocyte activation of oocytes matured in vitro.Results Sperm PLC-ζ protein deficiency was demonstrated by Western blot analysis and immunofluorescence and confirmed by reduced PLC-ζ bioactivity using an in vitro model of Ca 2+ release. Nevertheless, with this sperm and supplementation of Ca 2+ ionophore following ICSI, fertilization of four of six oocytes matured in vitro was obtained. In addition, four embryos underwent cleavage and two of them reached the blastocyst stage. Transfer of these blastocysts into the uterus led to a single pregnancy and live birth. Conclusions Deficiency of PLC-ζ in normal-appearing human sperm is associated with impaired Ca 2+ -dependent oocyte activation during ICSI. Under this condition, use of Ca 2+ ionophore following ICSI of oocytes matured in vitro improves embryo developmental competence, possibly through the activation of Ca 2+ -dependent mechanisms governing fertilization and preimplantation embryogenesis.

Section: Introductionmentioning

confidence: 86%

“…A second GnRH-antagonist ovarian stimulation with IVF/ ICSI was performed using Ca 2+ ionophore with ICSI to enhance Ca 2+ -induced oocyte activation, as previously reported [9,14,15]. This time, rhFSH and urinary gonadotropins were started at a higher dose of 375 IU sc daily for 2 days and then changed as clinically needed.…”

Section: Casementioning

confidence: 99%

Phospholipase C-zeta deficiency as a cause for repetitive oocyte fertilization failure during ovarian stimulation for in vitro fertilization with ICSI: a case report

Chithiwala

Lee

Hill

et al. 2015

“…There are seven cases reported of AOA after complete fertilization failure in normozoospermia [8][9][10][11][12]19]; in some the cause was not clearly identified [8,10], or was due to an oocyte factor [9]. Among those, six cases were treated using Ca 2+ ionophore [8,10,19]. In a case report of a normozoospermic patient [10], most sperm heads that appeared normal under light microscopy, exhibited abnormal nuclear vacuolization when analyzed by transmission electron microscopy (TEM).…”

Section: Icsi Failures In Normozoospermiamentioning

confidence: 99%

“…Assisted oocyte activation (AOA) with a Ca 2+ ionophore (A213187 or ionomycin), or strontium chloride (SrCl 2 ), direct microinjection of calcium, or electrical stimuli, have been shown to overcome fertilization failure after ICSI [6]. Ca 2+ ionophore is most widely used for AOA [7], whose most common indication is globozoospermia, with only seven cases of AOA reported with normozoospermic patients worldwide [8][9][10][11][12][13].…”

Section: Introductionmentioning

confidence: 99%

PLCζ disruption with complete fertilization failure in normozoospermia

Durbán¹,

Barragán²,

Colodrón³

et al. 2015

Purpose Intracytoplasmic sperm injection (ICSI) is widely used to achieve fertilization in the presence of severe male factor, resulting in high fertilization rates. Nevertheless, 1-3 % of couples experience complete fertilization failure after ICSI. When a male factor is identified, assisted oocyte activation (AOA) can help overcome fertilization failures. The objective of this study is to describe a case of repeated complete fertilization failures after ICSI with donor oocytes, and to investigate the molecular and functional aspects of phospholipase C zeta (PLCζ) protein in the patient semen. Methods The patient was a normozoospermic male who had previously fathered, through natural conception, four children by a different partner. Molecular and functional analysis of sperm-specific PLCζ in the patient and control samples by means of gene sequencing, immunocytochemistry, Western blot, mouse oocyte activation test (MOAT), and mouse oocyte calcium analysis (MOCA) were used.Results PLCζ expression levels and distribution were significantly disrupted, although MOAT and MOCA did not indicate a decrease in activation ability. Conclusions Normozoospermic males can have disrupted expression and distribution of PLCζ, and reduced activation ability after ICSI in human oocytes, despite their normal activation potential in functional testing using mouse oocytes. Discrepancy among molecular and functional data might exist, as mutations in the gene sequence may not be the only cause of alteration in PLCζ protein related to activation failures.

“…The reasons for this phenomenon was considered to be a partial or complete inability of the spermatozoa to activate the oocytes, deficiency of sperm protamine, or the inability of the oocytes to decondense spermatozoa [1][2][3]. When the oocytes were activated using electroporation [4,5], calcium ionophore [6][7][8][9][10][11][12][13][14], or calcium ionophore and puromycin [15,16], followed by ICSI in women whose oocytes failed to fertilize in previous IVF cycles, some of them could form pronuclei. Recently, several studies were reported that strontium chloride (SrCl 2 ) treatment on infertile patients resulted in successful pregnancies and deliveries [17][18][19].…”

Section: Introductionmentioning

confidence: 99%

Live birth after SrCl2 oocyte activation in previous repeated failed or low fertilization rates after ICSI of frozen-thawed testicular spermatozoa: case report

Kim

Choi

Yang

et al. 2012

Purpose To report a live birth resulting after strontium chloride (SrCl 2 ) oocyte activation in a couple with complete fertilization failure or low fertilization rates following intracytoplasmic sperm injection (ICSI) of frozen-thawed testicular spermatozoa. Methods The couple underwent ICSI of frozen-thawed testicular spermatozoa. After ICSI, the oocytes were artificially activated by SrCl 2 because the results of fertilization were not satisfactory in the previous cycles. The main outcome measures were fertilization, pregnancy, and birth. Results In the first and second cycles performed previously at another clinic, fertilization rates were 9.1 % and 0.0 %, respectively. In the third cycle, 31 metaphase II oocytes were retrieved. After sperm injection, all of the oocytes were stimulated using SrCl 2 for activation. Sixteen oocytes were fertilized (51.6 %), and a single embryo was transferred into the uterus on Day 3. A healthy girl weighing 2750 g was born at 40 weeks of gestation by caesarean section. Conclusions This result suggests that SrCl 2 could be useful for oocyte fertilization in case of repeated complete fertilization failure or low fertilization rates following ICSI of frozen-thawed testicular spermatozoa.