Levels of sterols including δ7-dehydrogenase isolated from the tissues of marine animal products (20 species) were determined on 1.5% OV-17 columm of gas-liquid chromatography. The composition showed that the mussels and clams contained various sterols in their tissues : cholesterol, brassicasterol. 24-methylenecholesterol with some minor components such as 22-trans-norcholesta-5,22-dien-3β-ol, 22-cis-dehydrocholesterol, 22-trans-dehydrocholesterol, desmosterol, 7-dehydrocholesterol, campesterol, stigmasterol, β-sitosterol, isofucosterol, and 7-dehydrocholesterol which could be converted into vitamin D3 in the skin tissue of animal was present in the muscle of oyster, Crassostrea gigas. On the other hand, the others including gastropoda were predominantly composed of cholesterol. The minor sterols such as 24-methylenecholesterol, stigmasterol and β-sitosterol in the fish intestines are supposed to be derived from dietary plankton. Cholesterol δ7-dehydrogenase which could convert cholesterol into δ7-dehydrogenase was present in the pickles of Tricurus haumela intestine.

Contents of vitamin D3 and 25-OH-vitamin D3 in marine animal products(20 species) were determined by HPLC. The isomers of vitamin D, D2 and D3, were not clearly separated on a reversed phase, μ Bonda Pak, with 20% methanol-acetonitrile, and on a normal phase, Zorbax SIL. with 0.4% isopropanol-hexane, but 25-OH-vitamin D2 and-D3 were separated on either μ Bonda Pak with 10% methanol-acetonitrile, or on Zorbax SIL with 2.2% isopropanol-hexane, respectively. Although levels of vitamin D3 and 25-OH-vitamin D3 varied remarkably according to species, their average value(fish : l,l87sim36,007 I.U/sample 100g, mussel : 58~1,706 I.U/sample 100g, pickle: 1,208~79,358 I.U/sample 100g) was greatly higher than that of meat(80~100 I.U/sample 100g) and dairy products(400~800 I.U/sample 100g). Fatty tissues of fish and pickled fish intestines contained high level of vitamin D3 and 25-OH-vitamin D3, while the clam and mussel known to have various kinds of sterol including δ7-sterol showed very low levels of vitamin D3 and its derivative.

The present study was undertaken to determine the optimal condition for parthenogenetic activation of rabbit oocytes by electric stimulation in vitro in an attempt to develop nuclear transplantation techniques for cloning mammalian embryos and animals. Freshly ovulated oocytes were collected from superovulated rabbits from 13 to 26 hrs. after hCG injection. The cumulus-free oocytes were activated parthenogetically by repeated stimuli of square direct electric pulses in O.3M mannitol solution. After applying electric stimulations of different voltages, pulse durations and pulse times, all of the oocytes were cultured in TCM-199 with 10% FCS for 96 hours in a 5% incubator, and their developmental potential in vitro was examined. The higher activation rate (68.9%) was achieved at the voltage of 2.0kv/cm, the pulse duration of 60 sec and three pulse times and the activation rate of 100% was achieved at the pulse duration of 100 and 200 sec, the voltage of 1.5kv/cm and three pulse times. Although the higher rates of activation of oocytes were achieved at 100 and 200 sec, none of them developed to blastocyst in vitro. The oocytes collected 18~20 hours post hCG injection showed the highest rate of activation and development to blastocyst in vitro than the oocytes collected 13~15 or 25~26 hours post hCG injection. Therefore, it can be suggested that the application of electric stimulation of 2.0kv/cm, 60 sec and three pulse times to the oocytes collected at 18~20 hours post hCG injection would be more beneficial for the parthenogenetic activation of oocytes in rabbits.

Nuclear transplantation techinque has been found to be the most potential and efficient method for producing a large number of genetically identical animals from a single embryo. The technical development of nuclear transplantation in mammals and its application to the production of cloned animals were reviewed. For the efficient and successful production of cloned embryos by nuclear transplantation, selection and micromanipulation of recipient eggs or embryos as capacious recipient cytoplasm, and benefitial preparation of multiple totipotent embryonic cells as donor nuclei, and also fusion technique are very critical. Recent works approaching to these critical points were introduced and discussed.