scholarly journals Time of Sperm Penetration into Rat Oocytes Matured In Vivo and In Vitro

1976 ◽  
Vol 47 (11) ◽  
pp. 654-658
Author(s):  
Koji NIWA ◽  
Masashi MIYAKE ◽  
Akira IRITANI ◽  
Yoshimasa NISHIKAWA
Keyword(s):  
Sperm Penetration

O-228 The SSRI antidepressant Sertraline inhibits CatSper calcium channels in human sperm

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
R Rahban ◽  
A Rehfeld ◽  
C Schiffer ◽  
C Brenker ◽  
D. Louise Egeberg Palme ◽  
...  
Keyword(s):  
In Vitro Study ◽  
Human Sperm ◽  
Fertilization Process ◽  
Acrosomal Exocytosis ◽  
Depth Analysis ◽  
Sperm Penetration ◽  
Viscous Media ◽  
Voltage Activation

Abstract Study question Do Selective Serotonin Reuptake Inhibitor (SSRI) antidepressants affect the function of human sperm? Summary answer The SSRI-antidepressant Sertraline (e.g. Zoloft) inhibits the sperm-specific Ca2+ channel CatSper and affects human sperm function in vitro. What is known already In human sperm, CatSper translates changes of the chemical microenvironment into changes of the intracellular Ca2+ concentration ([Ca2+]i) and swimming behavior. CatSper is promiscuously activated by oviductal ligands, but also by synthetic chemicals that might disturb the fertilization process. It is well known that SSRIs have off-target actions on Ca2+, Na+, and K+ channels in somatic cells. Whether SSRIs affect the activity of CatSper is, however, unknown. Study design, size, duration We studied the action of the seven drugs belonging to the most commonly prescribed class of antidepressants, SSRIs, on resting [Ca2+]i and Ca2+ influx via CatSper in human sperm. The SSRI Sertraline was selected for in-depth analysis of its action on steroid-, prostaglandin-, pH-, and voltage-activation of human CatSper. Moreover, the action of Sertraline on sperm acrosomal exocytosis and penetration into viscous media was evaluated. Participants/materials, setting, methods The activity of CatSper was investigated in sperm of healthy volunteers, using kinetic Ca2+ fluorimetry and patch-clamp recordings. Acrosomal exocytosis was investigated using Pisum sativum agglutinin (PSA) and image cytometry. Sperm penetration in viscous media was evaluated using the Kremer test. Main results and the role of chance Four SSRIs increased [Ca2+]i, two out of which also attenuated ligand-induced Ca2+ influx via CatSper. In contrast, Sertraline decreased [Ca2+]i and almost completely suppressed ligand-induced Ca2+ influx via CatSper. Remarkably, the drug was about four-fold more potent to suppress prostaglandin- versus steroid-induced Ca2+ influx. Sertraline also suppressed alkaline- and voltage-activation of CatSper, indicating that the drug directly inhibits human CatSper. Finally, Sertraline suppressed ligand-induced acrosome reaction and sperm penetration into viscous media. Limitations, reasons for caution This is an in vitro study. Future studies have to assess the physiological relevance in vivo. Wider implications of the findings The off-target action of Sertraline on CatSper in human sperm might impair the fertilization process. In a research setting, Sertraline may be used to selectively inhibit prostaglandin-induced Ca2+ influx. Trial registration number CRU326

scholarly journals Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro

Reproduction ◽  
2012 ◽  
Vol 144 (6) ◽  
pp. 687-697 ◽  
Author(s):  
J Beek ◽  
H Nauwynck ◽  
D Maes ◽  
A Van Soom
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Inhibitory Effect ◽  
Fertilization Rate ◽  
Quality Parameters ◽  
Fertilization In Vitro ◽  
Cumulus Oophorus ◽  
Sperm Penetration

In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 μM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1–TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

In Vitro Sperm Penetration Speed and its Relationship with In Vivo Bull Fertility

2004 ◽  
Vol 39 (6) ◽  
pp. 424-428 ◽  
Author(s):  
R Puglisi ◽  
D Balduzzi ◽  
A Galli
Keyword(s):  
Sperm Penetration

Relationship between bovine oocyte morphology and in vitro developmental potential

Zygote ◽  
2006 ◽  
Vol 14 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Masashi Nagano ◽  
Seiji Katagiri ◽  
Yoshiyuki Takahashi
Keyword(s):  
Small Diameter ◽  
Blastocyst Stage ◽  
Cortical Granules ◽  
Developmental Potential ◽  
Developmental Capacity ◽  
Sperm Penetration ◽  
Developmental Rates ◽  
Large Clusters

We investigated the relationship between the morphology of oocytes collected from small antral follicles and their developmental capacity. Immature oocytes were classified into seven groups and cultured in vitro for maturation (IVM), fertilization (IVF) and development to blastocysts (IVC). After IVF, sperm penetration and normal fertilization rates were higher in the oocytes whose cytoplasm appeared brown. The rate of polyspermy was highest in the oocytes whose cytoplasm was black. After IVC, the rates of cleavage and of development to the blastocyst stage were also higher in the brown oocytes. Although the oocytes with dark clusters in a pale cytoplasm showed lower cleavage rates, cleaved zygotes had high developmental rates the same as the oocytes with a brown cytoplasm. Transmission electron microscopy showed that the oocytes with a pale or black cytoplasm had organelles arranged differently from other oocytes before IVM. Most of the oocytes with a brown and homogeneous cytoplasm or small diameter had the characteristics of immature cytoplasm (large clusters of cortical granules) even after IVM. On the other hand, the brown oocytes with a dark zone at the periphery or with dark clusters showed the same arrangement of organelles as in vivo matured oocytes. The oocytes with a pale or black cytoplasm appeared to be degenerating and/or ageing. In conclusion, a dark ooplasm indicates an accumulation of lipids and good developmental potential, while a light-coloured ooplasm indicates a low density of organelles and poor developmental potential. A black ooplasm indicates ageing and low developmental potential.

Penetration of concanavalin-A-treated Chinese hamster oocytes by golden hamster spermatozoa in vitro, and chromosome analysis of hybrid 1-cell zygotes

Zygote ◽  
1996 ◽  
Vol 4 (3) ◽  
pp. 167-172 ◽  
Author(s):  
Seiji Watanabe ◽  
Hiroyuki Tateno ◽  
Yujiroh Kamiguchi
Keyword(s):  
Concanavalin A ◽  
Golden Hamster ◽  
Chinese Hamster ◽  
Chromosome Analysis ◽  
Con A ◽  
Positive Effects ◽  
Sperm Binding ◽  
Sperm Penetration

SummaryPretreatment of zona-free Chinese hamster (CH) oocytes with three kinds of lectin – concanavalin A (Con-A), phytohaemagglutinin-P (PHA) and wheat germ agglutinin (WGA) – was attempted in order to improve penetration by golden hamster (GH) spermatozoa in vitro. Con-A had no significant effect on penetration at 2 μg/ml, adequately facilitated oocyte-sperm fusion at 4 μg/ml, and caused excessive sperm binding and resultant severe polyspermy at 10 μg/ml. Neither PHA nor WGA had positive effects on sperm penetration at any concentrations (2–10 μg/ml) examined. Using the Con-A (4 μg/ml)pretreatment, high rates of interspecific fertilisation and subsequent chromosome analysis of hybrid 1-cell zygotes were achieved. Among 258 CH oocytes used, 212 (82.2%) were fertilised and 153 (72.2% of fertilised ova) developed to the first cleavage metaphase. Eventually, 132 CH-derived chromosome complements and 153 GH-derived ones were successfully karyoanalysed. Incidences of aneuploidy and structural anomaly were 3.1% and 2.3% in CH complements, and 1.4% and 6.5% in GH complements, respectively. These incidences were not significantly different from those obtained by intraspecific in vivo fertilisation, suggesting that our interspecific in vitro fertilisation system does not cause chromosome aberrations.

scholarly journals In vivo meiotic resumption, fertilization and early embryonic development in the bitch

Reproduction ◽  
2005 ◽  
Vol 130 (2) ◽  
pp. 193-201 ◽  
Author(s):  
K Reynaud ◽  
A Fontbonne ◽  
N Marseloo ◽  
S Thoumire ◽  
M Chebrout ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Accurate Determination ◽  
Germinal Vesicle ◽  
Specific Pattern ◽  
Cell Stage ◽  
Immature Oocytes ◽  
Sperm Penetration ◽  
Nuclear Stage

Early development in canine species follows a very specific pattern. Oocytes are ovulated at the germinal vesicle stage and meiotic resumption occurs in the oviduct. However, because of difficulties in the accurate determination of ovulation time and in the observation of oocyte nuclear stage by light microscopy, these early events have not been fully described. Moreover, the oocyte stage at which sperm penetration occurs is still uncertain since fertilization of immature oocytes has been reportedin vivoandin vitro. The aim of this study was to establish the exact timing ofin vivomeiotic resumption, fertilization and early embryo development in the bitch with reference to ovulation. Ovulation was first determined by ultrasonography, artificial inseminations were performed daily and oocytes/embryos were collected between 17 and 138 h after ovulation. After fixation and DNA/tubulin staining, the nuclear stage was observed by confocal microscopy. Of the 195 oocytes/embryos collected from 50 bitches, the germinal vesicle stage was the only one present until 44 h post-ovulation, and the first metaphase II stage was observed for the first time at 54 h. Sperm penetration of immature oocytes appeared to be exceptional (three out of 112 immature oocytes). In most cases, fertilization occurred from 90 h post-ovulation in metaphase II oocytes. Embryonic development was observed up to the eight-cell stage. No significant influence of bitch breed and age on ovulation rate, maturation and developmental kinetics was observed. However, some heterogeneity in the maturation/development process was observed within the cohort of oocytes/embryos collected from one bitch. In conclusion, the most peculiar aspect of the canine species remains oocyte meiotic maturation whereas fertilization follows the same pattern as in other mammals.

Maturation of hamster oocytes under chemically defined conditions and sperm penetration through the zona pellucida

Zygote ◽  
1996 ◽  
Vol 4 (3) ◽  
pp. 199-210 ◽  
Author(s):  
Seiji Kito ◽  
Barry Bavister
Keyword(s):  
Zona Pellucida ◽  
Sperm Concentration ◽  
Cumulus Cells ◽  
Defined Medium ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Sperm Penetration ◽  
Time Required

SummaryThis study aimed to achieve high frequencies of nuclear maturation and penetrability through the zona pellucida of hamster oocytes cultured under protein-free conditions. Completion of nuclear maturation by cumulus-intact, immature oocytes (79% metaphase II stage) was depressed (37% p < 0·05) by adding four amino acids (glutamine, isoleucine, methionine and phenylalanine) reported necessary for nuclear maturation of cumulus-free oocytes. Following in vitro maturation, cumulus cells were removed and oocytes were inseminated with capacitated sperm, but after 6 h sperm:egg co-incubation, only 24% of in vitro matured oocytes were penetrated compared with 60% of in vivo matured oocytes (p < 0·05). Time required for zona lysis by α-chymotrypsin was not significantly different among in vitro and in vivo matured oocytes and 1-cell embryos. Addition to the maturation medium of soybean trypsin inhibitor or fetuin, both known to inhibit the zona reaction in vitro, did not improve penetrability of in vitro matured oocytes, implying that in hamsters, unlike other rodent species, a premature zona reaction is unlikely to be responsible for inhibiting sperm penetration. When oocytes were incubated with 20% periovulatory oviductal fluid (OF) for another 3 h after maturation, penetration was significantly improved (60% vs 37% with and without OF, respectively; p < 0·05), but was not equivalent to penetration of in vivo matured follicular oocytes similarly treated with OF (84%, p < 0·05)However, zona penetration was further improved by increasing sperm concentration from 1·0 × 104 (66%) to 5·0 or 10·0 × 104 sperm/ml (89%, p < 0·05). This study shows that nuclear maturation of hamster oocytes can occur in chemically defined medium, and indicates that a deficiency in the zona of in vitro matured oocytes can be overcome by preincubation with OF and insemination at high sperm conccentration.

scholarly journals The antidepressant Sertraline inhibits CatSper Ca2+ channels in human sperm

2021 ◽  
Vol 36 (10) ◽  
pp. 2638-2648
Author(s):  
Rita Rahban ◽  
Anders Rehfeld ◽  
Christian Schiffer ◽  
Christoph Brenker ◽  
Dorte Louise Egeberg Palme ◽  
...  
Keyword(s):  
Human Sperm ◽  
German Research Foundation ◽  
Fertilization Process ◽  
Acrosomal Exocytosis ◽  
Depth Analysis ◽  
Sperm Penetration ◽  
Viscous Media ◽  
Voltage Activation

Abstract STUDY QUESTION Do selective serotonin reuptake inhibitor (SSRI) antidepressants affect the function of human sperm? SUMMARY ANSWER The SSRI antidepressant Sertraline (e.g. Zoloft) inhibits the sperm-specific Ca2+ channel CatSper and affects human sperm function in vitro. WHAT IS KNOWN ALREADY In human sperm, CatSper translates changes of the chemical microenvironment into changes of the intracellular Ca2+ concentration ([Ca2+]i) and swimming behavior. CatSper is promiscuously activated by oviductal ligands, but also by synthetic chemicals that might disturb the fertilization process. It is well known that SSRIs have off-target actions on Ca2+, Na+ and K+ channels in somatic cells. Whether SSRIs affect the activity of CatSper is, however, unknown. STUDY DESIGN, SIZE, DURATION We studied the action of the seven drugs belonging to the most commonly prescribed class of antidepressants, SSRIs, on resting [Ca2+]i and Ca2+ influx via CatSper in human sperm. The SSRI Sertraline was selected for in-depth analysis of its action on steroid-, prostaglandin-, pH- and voltage-activation of human CatSper. Moreover, the action of Sertraline on sperm acrosomal exocytosis and penetration into viscous media was evaluated. PARTICIPANTS/MATERIALS, SETTING, METHODS The activity of CatSper was investigated in sperm of healthy volunteers, using kinetic Ca2+ fluorimetry and patch-clamp recordings. Acrosomal exocytosis was investigated using Pisum sativum agglutinin and image cytometry. Sperm penetration in viscous media was evaluated using the Kremer test. MAIN RESULTS AND THE ROLE OF CHANCE Several SSRIs affected [Ca2+]i and attenuated ligand-induced Ca2+ influx via CatSper. In particular, the SSRI Sertraline almost completely suppressed Ca2+ influx via CatSper. Remarkably, the drug was about four-fold more potent to suppress prostaglandin- versus steroid-induced Ca2+ influx. Sertraline also suppressed alkaline- and voltage-activation of CatSper, indicating that the drug directly inhibits the channel. Finally, Sertraline impaired ligand-induced acrosome reaction and sperm penetration into viscous media. LIMITATIONS, REASONS FOR CAUTION This is an in vitro study. Future studies have to assess the physiological relevance in vivo. WIDER IMPLICATIONS OF THE FINDINGS The off-target action of Sertraline on CatSper in human sperm might impair the fertilization process. In a research setting, Sertraline may be used to selectively inhibit prostaglandin-induced Ca2+ influx. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the Swiss Centre for Applied Human Toxicology (SCAHT), the Département de l’Instruction Publique of the State of Geneva, the German Research Foundation (CRU326), the Interdisciplinary Center for Clinical Research, Münster (IZKF; Str/014/21), the Innovation Fund Denmark (grant numbers 14-2013-4) and the EDMaRC research grant from the Kirsten and Freddy Johansen’s Foundation. The authors declare that no conflict of interest could be perceived as prejudicing the impartiality of the research reported. TRIAL REGISTRATION NUMBER NA.

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