Methods 31 Matings to Supply Fertilized Eggs

In our laboratory, young and immature Sprague Dawley female rats (see Note 1) of 5-wk-old are used for superovulation by the administration of gonadotrophins as follows:

1. At 1200 h, inject 5-10 sexually mature female rats intraperitoneally with 10 IU of FSH {see Note 2).

2. 56 h later, inject the females intraperitoneally with 10 IU of hCG (see Note 2) before placing each with a stud male (see Note 2)

3. In the morning, the females are subjected to a vaginal smear (see Section 3.3 ) to check for the presence of sperm, which is an indication of successful copulation. In rats, despite successful mating, the copulatory plug sometimes has been lost by the next morning. Hence, only females that have a positive sperm smear (see Section 3.3.) are used to supply fertilized eggs. Some 50-80% of superovulated females should mate

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