Assessment of Oocyte Competence

Abstract

Methods are provided for evaluating an oocyte for fertilization and implantation. For example, methods are provided for determining whether an oocyte expresses, or does not express, one or more of a group of markers identified as differently expressed between chromosomally normal and chromosomally abnormal oocytes. Also provided, for example, are methods for determining whether a cumulus cell expresses, or does not express, one or more of a group of markers identified as differently expressed between cumulus cells associated with chromosomally normal oocytes and cumulus cells associated with chromosomally abnormal oocytes. Methods are provided for the detection of marker expression of differentially expressed genes at the RNA level, as well as at the protein level.

1 . A method of evaluating the competence of a mammalian oocyte for implantation comprising:
 (i) determining in a sample the level of marker expression of at least one nucleic acid selected from the group of nucleic acids exemplified by SEQ ID NOS:1-92, 252, and 351, and   (ii) comparing the level of marker expression in the sample with a control or reference standard, wherein detecting differential marker expression between the sample and the control is indicative of the competence of the oocyte for implantation.   
     
     
         2 . The method of  claim 1 , wherein the sample is derived from an oocyte. 
     
     
         3 . The method of  claim 2 , wherein the control or reference standard is derived from one of the group consisting of: an oocyte competent for implantation, a chromosomally normal oocyte, an oocyte not competent for implantation, and a chromosomally abnormal oocyte. 
     
     
         4 . The method of  claim 1 , wherein the sample is derived from follicular fluid. 
     
     
         5 . The method of  claim 4 , wherein the control or reference standard is derived from one of the group consisting of: follicular fluid associated with an oocyte competent for implantation, follicular fluid associated with a chromosomally normal oocyte, follicular fluid associated with an oocyte not competent for implantation and follicular fluid associated with a chromosomally abnormal oocyte. 
     
     
         6 . The method of  claim 1 , wherein the sample is derived from culture medium. 
     
     
         7 . The method of  claim 6 , wherein the control or reference standard is derived from one of the group consisting of: culture medium associated with an oocyte competent for implantation, culture medium associated with a chromosomally normal oocyte, culture medium associated with an oocyte not competent for implantation, and culture medium associated with a chromosomally abnormal oocyte. 
     
     
         8 . The method of  claim 1 , wherein the level of marker expression determined in the sample is at least 20% different from the level of marker expression determined in the control or reference standard. 
     
     
         9 . The method of  claim 1 , wherein the level of marker expression is detected by at least one of the group consisting of nucleic acid microarray, Northern blot, and reverse transcription PCR. 
     
     
         10 . The method of  claim 1 , wherein the level of marker expression is detected by at least one of the group consisting of Western blot, enzyme-linked immunosorbent assay, protein microarray and FACS analysis. 
     
     
         11 . The method of  claim 1 , wherein the mammalian oocyte is of a domesticated mammal. 
     
     
         12 . The method of  claim 1 , wherein the mammalian oocyte is of a human. 
     
     
         13 . A method of evaluating the competence of a mammalian oocyte for implantation comprising:
 (i) determining in a sample the level of marker expression of at least one nucleic acid selected from the group of nucleic acids exemplified by SEQ ID NOS:183-282, 252, and 351, and   (ii) comparing the level of marker expression in the sample with a control or reference standard, wherein detecting differential marker expression between the sample and the control is indicative of the competence of an oocyte for implantation.   
     
     
         14 . The method of  claim 13 , wherein the sample is derived from a cumulus cell. 
     
     
         15 . The method of  claim 14 , wherein the control or reference standard is derived from one of the group consisting of: a cumulus cell associated with an oocyte competent for implantation, a cumulus cell associated with a chromosomally normal oocyte, a cumulus cell associated with an oocyte not competent for implantation and a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         16 . The method of  claim 13 , wherein the sample is derived from follicular fluid. 
     
     
         17 . The method of  claim 16 , wherein the control or reference standard is derived from one of the group consisting of: follicular fluid associated with a cumulus cell associated with an oocyte competent for implantation, follicular fluid associated with a cumulus cell associated with a chromosomally normal oocyte, follicular fluid associated with a cumulus cell associated with an oocyte not competent for implantation and follicular fluid associated with a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         18 . The method of  claim 13 , wherein the sample is derived from culture medium. 
     
     
         19 . The method of  claim 18 , wherein the control or reference standard is derived from one of the group consisting of; culture medium associated with a cumulus cell associated with an oocyte competent for implantation, culture medium associated with a cumulus cell associated with a chromosomally normal oocyte, culture medium associated with a cumulus cell associated with an oocyte not competent for implantation, and culture medium associated with a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         20 . The method of  claim 13 , wherein the level of marker expression determined in the sample is at least 20% different from the level of marker expression determined in the control or reference standard. 
     
     
         21 . The method of  claim 13 , wherein the level of marker expression is detected by at least one of the group consisting of nucleic acid microarray, Northern blot, and reverse transcription PCR. 
     
     
         22 . The method of  claim 13 , wherein the level of marker expression is detected by at least one of the group consisting of Western blot, enzyme-linked immunosorbent assay, protein microarray and FACS analysis. 
     
     
         23 . The method of  claim 13 , wherein the mammalian oocyte is of a domesticated mammal. 
     
     
         24 . The method of  claim 13 , wherein the mammalian oocyte is of a human. 
     
     
         25 . A method of evaluating the competence of a mammalian oocyte for fertilization comprising:
 (i) determining in a sample the level of marker expression of at least one nucleic acid selected from the group of nucleic acids exemplified by SEQ ID NOS:1-92, 252, and 351, and   (ii) comparing the level of marker expression in the sample with a control or reference standard, wherein detecting differential marker expression between the sample and the control is indicative of the competence of the oocyte for implantation.   
     
     
         26 . The method of  claim 25 , wherein the sample is derived from an oocyte. 
     
     
         27 . The method of  claim 26 , wherein the control or reference standard is derived from one of the group consisting of: an oocyte competent for fertilization, a chromosomally normal oocyte, an oocyte not competent for fertilization, and a chromosomally abnormal oocyte. 
     
     
         28 . The method of  claim 25 , wherein the sample is derived from follicular fluid. 
     
     
         29 . The method of  claim 28 , wherein the control or reference standard is derived from one of the group consisting of: follicular fluid associated with an oocyte competent for fertilization, follicular fluid associated with a chromosomally normal oocyte, follicular fluid associated with an oocyte not competent for fertilization, and follicular fluid associated with a chromosomally abnormal oocyte. 
     
     
         30 . The method of  claim 25 , wherein the sample is derived from culture medium. 
     
     
         31 . The method of  claim 30 , wherein the control or reference standard is derived from one of the group consisting of: culture medium associated with an oocyte competent for fertilization, culture medium associated with a chromosomally normal oocyte, culture medium associated with an oocyte not competent for fertilization, and culture medium associated with a chromosomally abnormal oocyte. 
     
     
         32 . The method of  claim 25 , wherein the level of marker expression determined in the sample is at least 20% different from the level of marker expression determined in the control or reference standard. 
     
     
         33 . The method of  claim 25 , wherein the level of marker expression is detected by at least one of the group consisting of nucleic acid microarray, Northern blot, and reverse transcription PCR. 
     
     
         34 . The method of  claim 25 , wherein the level of marker expression is detected by at least one of the group consisting of Western blot, enzyme-linked immunosorbent assay, protein microarray and FACS analysis. 
     
     
         35 . The method of  claim 25 , wherein the mammalian oocyte is of a domesticated mammal. 
     
     
         36 . The method of  claim 25 , wherein the mammalian oocyte is of a human. 
     
     
         37 . A method of evaluating the competence of a mammalian oocyte for fertilization comprising:
 (i) determining in a sample the level of marker expression of at least one nucleic acid selected from the group of nucleic acids exemplified by SEQ ID NOS:183-282, 252, and 351, and   (ii) comparing the level of marker expression in the sample with a control or reference standard, wherein detecting differential marker expression between the sample and the control is indicative of the competence of an oocyte for fertilization.   
     
     
         38 . The method of  claim 37 , wherein the sample is derived from a cumulus cell. 
     
     
         39 . The method of  claim 38 , wherein the control or reference standard is derived from one of the group consisting of: a cumulus cell associated with an oocyte competent for fertilization, a cumulus cell associated with a chromosomally normal oocyte, a cumulus cell associated with an oocyte not competent for fertilization, and a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         40 . The method of  claim 37 , wherein the sample is derived from follicular fluid. 
     
     
         41 . The method of  claim 40 , wherein the control or reference standard is derived from one of the group consisting of: follicular fluid associated with a cumulus cell associated with an oocyte competent for fertilization, follicular fluid associated with a cumulus cell associated with a chromosomally normal oocyte, follicular fluid associated with a cumulus cell associated with an oocyte not competent for fertilization, and follicular fluid associated with a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         42 . The method of  claim 37 , wherein the sample is derived from culture medium. 
     
     
         43 . The method of  claim 42 , wherein the control or reference standard is derived from one of the group consisting of: culture medium associated with a cumulus cell associated with an oocyte competent for fertilization, culture medium associated with a cumulus cell associated with a chromosomally normal oocyte, culture medium associated with a cumulus cell associated with an oocyte not competent for fertilization and culture medium associated with a cumulus cell associated with a chromosomally abnormal oocyte. 
     
     
         44 . The method of  claim 37 , wherein the level of marker expression determined in the sample is at least 20% different from the level of marker expression determined in the control or reference standard. 
     
     
         45 . The method of  claim 37 , wherein the level of marker expression is detected by at least one of the group consisting of nucleic acid microarray, Northern blot, and reverse transcription PCR. 
     
     
         46 . The method of  claim 37 , wherein the level of marker expression is detected by at least one of the group consisting of Western blot, enzyme-linked immunosorbent assay, protein microarray and FACS analysis. 
     
     
         47 . The method of  claim 37 , wherein the mammalian oocyte is of a domesticated mammal. 
     
     
         48 . The method of  claim 37 , wherein the mammalian oocyte is of a human. 
     
     
         49 . An array of nucleic acid probes immobilized on a solid support, the array comprising:
 a probe set comprising a plurality of probes, each probe comprising a segment of at least twenty nucleotides exactly complementary to a subsequence of a set of reference sequences, wherein the set of reference sequences comprises SEQ ID NOS:1-92, 252, and 351.   
     
     
         50 . An array of nucleic acid probes immobilized on a solid support, the array comprising:
 a probe set comprising a plurality of probes, each probe comprising a segment of at least twenty nucleotides exactly complementary to a subsequence of a set of reference sequences, wherein the set of reference sequences comprises SEQ ID NOS:183-282, 252, and 351.