In Vitro Fertilization: Increased Vla (Very Late Antigen) Integrins and Fibronectin After Acrosome Reaction

Small South American rodents of the genus Calomys have been used extensively for virology and ecological research. Previous studies have demonstrated that Calomys musculinus and Calomys laucha have a relatively short oestrous cycle and that superovulation and parthenogenetic activation can be induced. The purpose of this study was to determine the requirements for in vitro manipulation of the male gamete and in vitro fertilization. Two culture media and different concentrations of spermatozoa were tested for their ability to support sperm motility, hyperactivation and the acrosome reaction. The ability of capacitated Calomys spermatozoa to penetrate zona-free hamster eggs was also evaluated. In vitro fertilization was assessed by examining attachment and binding to the zona pellucida, second polar body extrusion, pronucleus formation and the fertilizing sperm tail. The results of the study showed that: (i) Tyrode's albumin lactate pyruvate (TALP) medium was more effective than T6 medium for maintaining sperm motility in vitro; (ii) hyperactivation was achieved with TALP but not with T6; (iii) the acrosome reaction was easily distinguished by light microscopy and depends on time and sperm concentration; (iv) capacitated spermatozoa are able to penetrate zona-free hamster eggs; and (v) superovulated oocytes can be fertilized in vitro. This is the first report of capacitation and in vitro fertilization for Calomys sp. These results provide opportunities to use C. musculinus and C. laucha as new laboratory animals for research into reproductive biology.

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Andrology: A new predictive test for in-vitro fertilization based on the induction of sperm acrosome reaction by N-acetylglucosamine-neoglycoprotein

Human Reproduction ◽

10.1093/oxfordjournals.humrep.a136168 ◽

1995 ◽

Vol 10 (7) ◽

pp. 1751-1756 ◽

Cited By ~ 15

Author(s):

Adriano Brandelli ◽

Patricia V. Miranda ◽

María G. Añón-Vazquez ◽

Clara I. Marín-Briggiler ◽

Claudia Sanjurjo ◽

...

Keyword(s):

In Vitro Fertilization ◽

Acrosome Reaction ◽

Predictive Test ◽

Vitro Fertilization ◽

Sperm Acrosome Reaction ◽

Sperm Acrosome

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106EVALUATION OF ADDITION OF REDUCED GLUTATHIONE TO COOLING MEDIUM ON IN VITRO FERTILITY AND ACROSOME REACTION IN BOAR SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab106 ◽

2004 ◽

Vol 16 (2) ◽

pp. 175

Author(s):

C. Matás ◽

J. Gadea ◽

F. García-Vázquez ◽

J.C. Gardón ◽

S. Cánovas

Keyword(s):

In Vitro Fertilization ◽

Acrosome Reaction ◽

Reduced Glutathione ◽

Membrane Integrity ◽

Calcium Ionophore ◽

Egg Yolk ◽

Ros Generation ◽

Vitro Fertilization ◽

Cooling Medium

The process of cooling to 5°C prior to freezing produces physical and chemical stress on the sperm membrane associated with oxidative stress and reactive oxygen species (ROS) generation that reduces sperm viability and fertilizing ability. The addition of antioxidants to cooling medium could prevent the formation of ROS and improve the seminal parameters. The aim of these experiments was to investigate the effects of addition of reduced glutathione (GSH) to cooling extenders on (1) plasma membrane integrity, (2) acrosome reaction induction by ionophore A 23187 or progesterone, and (3) in vitro fertilization. Ejaculate-rich fractions from three mature pietrain boars were diluted in Beltsville Thaw Solution (BTS) extender and cooled to 15°C over 2h (group C). Thereafter, sperm were centrifuged and diluted in lactose/egg-yolk extender with 0mM (group 0), 1mM (group 1) or 5mM (group 5) of GSH, cooled to 5°C over 2h. The acrosome reaction was then induced by 1μM calcium ionophore or 10μM progesterone in TALP medium and incubated in 5% CO2, 38.5°C for 30 or 45min, respectively. Membrane integrity was evaluated by propidium iodide, and acrosomal status was monitored by means of FITC-labeled peanut agglutinin. Finally, in vitro fertilization was performed with these four spermatozoa groups as described previously (Matás et al. 2003 Reproduction 125, 133–141). ANOVA analysis revealed that the addition of GSH had no effect on the membrane integrity (ranged 58.8 to 66.9) or acrosome reaction induction (ranged 24.3 to 28.2, and 55.7 to 41.4 for progesterone and calcium ionophore, respectively). However, the results of the penetration assay revealed that the cooling affected the penetration rate and the number of sperm per oocyte (Table 1), and this assay is better than the others to predict changes in the spermatozoa functionality (Gadea J and Matás C 2000 Theriogenology 54, 1343–1357). In conclusion, the cooling process affects the in vitro fertilization, but the addition of GSH to the medium did not influence the parameters studied. Supported by AGL2000-0485-CO2-01. Table 1 Homologous in vitro penetration

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Capacitation and the acrosome reaction in marsupial spermatozoa

Reproduction Fertility and Development ◽

10.1071/rd9960595 ◽

1996 ◽

Vol 8 (4) ◽

pp. 595 ◽

Cited By ~ 9

Author(s):

KE Mate ◽

JC Rodger

Keyword(s):

In Vitro Fertilization ◽

Embryo Development ◽

Acrosome Reaction ◽

Culture Conditions ◽

Current Understanding ◽

Critical Event ◽

Vitro Fertilization ◽

Australian Species

Although yet to be established definitively, it appears that marsupial spermatozoa require a process of capacitation and that the mechanisms involved may be quite different between the Australian and American species. For Australian species, failure to induce this functional event in culture has meant that in vitro fertilization (IVF) is yet to be achieved. However, in the American species with paired spermatozoa, IVF and subsequent embryo development have been obtained under quite simple culture conditions. Our understanding of the interactions of marsupial spermatozoa with the female tract, and in particular the oviduct, the most likely site of capacitation, is discussed. Although the acrosome reaction (AR) is an equally critical event in marsupial fertilization it appears to be regulated quite differently. The uniquely stable character of the marsupial acrosome is examined as well as our current understanding of the regulation of the marsupial sperm AR in vivo and in vitro.

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