Analysis of Fertilization Phenomenon Using the Patch Clamp Method.

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This research aimed at the analysis of fertilization phenomenon using the patch clamp method which is newly developed method in electro-physiology area.(1) Study of golden hamster ovumWe injected PMS and hCG in subcutaneous tissue of golden hamster. Hyperstimulated ova were collected using microscope and incubated in trypsin and hyaronidaze contained solution. Collected zona free ovum was attached with patch clamp electrode, and on-off phenomena of calcium ion channel were recorded and analyzed. After study of calcium ion channel in sperm free solution, sperm of golden hamster were added into solution. Frequency of on-off phenomena of calcium ion channel were increased after penetration of sperm into ovum under phase contrast microscope. These data are already speculated in other methods, but it is important to get these data in direct method.(2) Study of human ovumBecause studies of fertilization of human ovum are restricted under Japanese Obstetrics & gynecology Association now, the precise studies of human ovum cannot be done. We examined human ova which were collected in gynecological operations under permission of patients. Zona free ovum which was obtained after treatment of trypsin and hyaronidaze was attached with patch clamp electrode. Frequency of on-off phenomena of calcium ion channel was low in follicular phase, but in luteal phase it was high frequency. These data are newly found out.