Oocyte activation by sperm components

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 5/06 (2006.01) A61D 19/00 (2006.01) C07K 2/00 (2006.01) C07K 14/47 (2006.01) C12N 5/08 (2006.01)

Patent

CA 2354735

The invention provides a microinsertion-based, trans-complementation method of in vitro oocyte activation useful to identify properties of sperm-borne oocyte- activating factor(s) (SOAF) and ooplasmic interactions with sperm components at fertilization. The invention provides at least one detergent-insoluble heat- sensitive component of the perinuclear matrix of a spermatozoon (SOAFm) that acts coordinately with at least one heat-stable submembrane sperm component. SOAFm is solubilized in vitro (SOAFs) under reducing conditions similar to those encountered in the ooplasm. By the method of the invention, the failure of heat-inactivated demembranated sperm heads to activate an egg is rescued by coinsertion of SOAFs into an oocyte. SOAFs is protease-sensitive and is liberated from demembranated heads in a temperature-dependent manner that inversely correlates with the ability of demembranated sperm heads to activate oocytes.

L'invention concerne une technique d'activation in vitro d'un ovocyte reposant sur la microinsertion et la trans-complémentation, permettant d'identifier les propriétés des facteurs activateurs d'ovocyte spermatiques (SOAF) et les interactions ovoplasmiques avec les composants spermatiques lors de la fécondation. Cette invention concerne au moins un composant non soluble dans les détergents et thermosensible de la matrice perinucléaire d'un spermatozoïde (SOAF¿m?) qui agit en coordination avec au moins un composant spermatique sous-membranaire thermostable. Le SOAF¿m? est solubilisé in vitro (SOAF¿s?) dans des conditions de réduction semblables à celles qui prévalent dans l'ovoplasme. Grâce à cette technique, un échec dans l'activation d'un oeuf par des têtes spermatiques thermo-inactivées et démembranées peut être rattrapé par la co-insertion de SOAF¿s? dans un ovocyte. Ce SOAF¿s? est sensible aux protéases et est libéré des têtes démembranées en fonction de la température, d'une manière inversement proportionnelle à la capacité des têtes démembranées d'activer les ovocytes.

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