Analysis of the function of the reproductive and immune system an approach by the embryo manipulation

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To investigate the reproductive and immune functions of transgenic mice, a linear DNA fragment encoding the regulatory region of mouse metalothionein and the strucural gene of human growth hormone was microinjected into the male pronucleus of C57BL/6 ova. After injection the ova were cultured overnight and transplanted into pseudopregnant oviducts. About 150 copies of human growth hormone gene was detected by the dot-blot gybridization and northern hybridization in the genomic DNA of a mouse born alive. The rates of survival in culture, implantation, and birth were normal, but the rate of integration was still low. For the use of transgenic mice as a protein synthetic system, we cloned cDNA for mouse casein 44k and whey acidic protein. It was also suggested in this series of study that the absence of a cytoplasmic factor or factors is involved in "two-cell block" in addition to an oviduct factor as has been suggested. Gnotobiote mice were also produced which have flora similar to the human flora to analyze their effects on the immune and dietary systems. The mode of action of prolactin is one the main subjects to study in this program because prolactin is a key hormone which regulates reproductive and immune function of animals. Macrophages are involved in the regulation of both immune and reproductive systems. Prolactin receptor was partially purified from rabbit mammary glands, and monoclonal antibodies were raised against it. We obtained two clones which recognized the recepto and inhibited prolactin binding. Using these antibodies the presence of two classes of receptor sites was suggested. In rat corpora lutea, prolactin inhibits the activity of 20<alpha>-hydroxysteroid dehydrogenase, which also was partially purified. This enzyme preparation can be used to clarified the mode of luteotrophic action of prolactin in corpora lutea. Transgenic mice and gnotobiote mice will be used as tools to anlyze the reproductive and immune systems in future.