Phospholipase c zeta mediated oocyte activation, compositions for use therein, and assays for detecting and identifying agents for treating male infertility

A method of detecting male infertility comprising:

• (i) determining a patient's phospholipase C zeta level by measuring the level of phospholipase C zeta in a sperm sample from the patient;
• (ii) comparing said patient's phospholipase C zeta level with a reference value, wherein said reference value is the minimum level of phospholipase C zeta observed in essentially all sperm samples from one or more control subjects known to exhibit normal fertility;
• (iii) identifying the patient as having male infertility if said patient's phospholipase C zeta level is significantly lower than the reference value;
• wherein said sperm sample from the patient and said sperm samples from one or more control subjects comprise sperm cells or sperm progenitor cells.

A method of detecting male infertility comprising:

• (i) determining the reference spatial localization of phospholipase C zeta protein, wherein said reference spatial localization of phospholipase C zeta protein is the spatial localization of phospholipase C zeta protein observed in the majority of sperm from the majority of control subjects known to have normal fertility, wherein said control subjects known to have normal fertility are of the same species as said patient;
• (ii) determining fraction of patient's sperm showing spatial localization of phospholipase C zeta protein comparable to the reference spatial localization of phospholipase C zeta protein;
• and
• (iii) identifying the patient as having male infertility if said fraction of patient's sperm showing spatial localization of phospholipase C zeta protein comparable to the reference spatial localization of phospholipase C zeta protein is significantly lower than 100%.

A method of treating male infertility, comprising:

• (i) contacting an oocyte in vitro with sperm from a patient suspected or known to have male infertility;
• (ii) introducing into or contacting said oocyte and/or sperm with a compatible phospholipase C zeta protein or a functional variant or fragment thereof or a nucleic acid sequence encoding any of the foregoing, or inducing said oocyte or sperm to express a phospholipase C zeta protein or a fragment or variant thereof;
• wherein steps (i) and (ii) may be effected in either order or simultaneous and result in an activated fertilized oocyte;
• and
• (iii) thereafter, implanting said activated, fertilized oocyte in a suitable host or culturing said activated, fertilized oocyte in vitro, whereby said oocyte develops into an embryo.

A method of detecting male infertility, comprising:

• (i) identifying non-functional allelic variants of phospholipase C zeta;
• (ii) determining partial or complete genomic sequences of a patient's phospholipase C zeta genes;
• and
• (iii) identifying the patient as having male infertility if said partial or complete genomic sequences of a patient's phospholipase C zeta genes are identical or similar to said non-functional allelic variants of phospholipase C zeta, or if said genomic sequences of a patient's phospholipase C zeta genes encode allelic variants of phospholipase C zeta having mutations that encode truncated phospholipase C zeta protein, or possess mutations within splice donor or splice acceptor sequences, or encode mutations within catalytic residues, or any combination thereof.

A method of identifying a composition useful for treatment of male infertility, comprising:

• (i) contacting one or more cells with a candidate composition;
• (ii) monitoring the intracellular calcium concentration of said one or more cells;
• and
• (iii) thereafter, identifying a composition as useful for treatment of male infertility if fluctuations of said intracellular calcium concentration are observed that are similar in frequency and amplitude to the fluctuations of intracellular calcium concentration observed when cells of the same type as said one or more cells are contacted with phospholipase C zeta or a nucleic acid sequence encoding.