생쥐 Age에 따른 난자-난구 복합체의 분산과 성숙에 관한 연구

이원교,권혁방,Lee, Won-Kyo,Kwon, Hyuk-Bang 대한생식의학회 1986 Clinical and Experimental Reproductive Medicine Vol.13 No.2

In order to know when the cumulus cells of mouse follicles get ability to expand in vitro, the oocyte cumulus complexes obtained from different growing ages of mice were cultured in the medium containing HCG and their rate of expansion were observed and at the same time their maturation rate was examined. The growth of follicles was also checked by histological method. It was impossible to isolate the oocyte-cumulus complexes from 13 or 15 days old mouse ovaries. The oocyte-cumulus complexes collected from 17 days old mouse were partially induced to expanded by HCG, and from 19 days, most of the complexes were induced to full expansion. The rate of cumulus cell expansion by HCG and the oocyte maturation increased steadly during the growing ages to adult. Thus, the time for follicles to get competence for expansion and maturation seems to be closely related. Antral follicles were appeared from 17 days old mice and Graafian follicles were seen from 21 days old mice. The competence for cumulus expansion increased during follicle growth up to 21 days old mice.

배양중에 있는 생쥐 여포난자(濾胞卵子)의 핵붕괴(核崩壞)(Germinal Vesicle Break-down)에 미치는 Progesterone의 영향에 관하여

조완규,권혁방,정순오,Cho, Wan-Kyoo,Kwon, Hyuk-Bang,Chung, Soon-O 대한생식의학회 1974 Clinical and Experimental Reproductive Medicine Vol.1 No.1

In the present studies, effect of progesterone on the germinal vesicle break-down of the mouse oocytes cultured in the micro tube was investigated. The results obtained are as follows: As dose of progesterone in the medium rose, accordingly the break-down of the germinal vesicle was suppressed. It was found that $ED_{50}$ was 15.7 ${\mu}g$/ml, and $ED_{90}$ 60.7 ${\mu}g$/ml of progesterone. The dose suppressing the oocyte maturation was apparently higher than that on the rabbit or on the mouse embryonal development. The inhibiting effect of progesterone on the GVBD was reversible. The germinal vesicle of the oocytes were broken down immediately in the medium upon removal of the hormone. Progesterone stops meiosis at any stage upon administration, while dbe AMP or theophylline supresses only the break-down of the nuclear membrane. Recovering of the meiotic division of the oocytes once exposed to progesterone was delayed a little. The inhibiting action of progesterone was not altered by adding more pyruvate or in the presence of higher concentration of the mineral ions in the culture medium.

인공배양한 생쥐 난자;난구복합체의 전자현미경적 연구

이기숙,김종덕,권혁방,Lee, Gy-Soog,Kim, Jong-Duk,Kwon, Hyuk-Bang 대한생식의학회 1990 Clinical and Experimental Reproductive Medicine Vol.17 No.2

These experiments were performed to know ultrastructural changes of the cumulus expansion in virot. SEM:In expanded oocyte-cumulus complex, the cell surface are characterized by the presence of many evaginations:they are relatively short and round shape. The mucous extracellular material were deposited between cumulus cells. TEM:In compact cumulus cells, golgi apparatus and rough endoplasmic reticulum developed. In expanded cumulus cells, rough endoplasmic reticulum decreased and the smooth endoplasmic reticulum increased. Also, there were numbers of mitochondria. Extracellular mucous material which is presumed to be hyaluronic acid appears when cumulus cell were expanded. In expanded cumulus cell, numbers of smooth endoplasmic reticulum help cumulus cell to develop in steroidogenic cell.

Gonadotropin Bioactivity and Steroids in Ovarian Follicle Matured by Hyperstimulation

윤용달,전은현,김문규,권혁방,Yoon, Yong-Dal,Chun, Eun-Hyun,Kim, Moon-Kyoo,Kwon, Hyuk-Bang The Korean Society for Reproductive Medicine 1989 Clinical and Experimental Reproductive Medicine Vol.16 No.2

본 연구는 생식주기중 폐쇄여포액내에서 생물학적, 면역학적 특성을 나타내는 GTH 의 변화를 조사하고 steroid hormone과의 상관관계를 조사하며 국부조절인자로서 의 GTH의 역 활을 조사하고자 하였다. 가임기간중 215개의 여포와 IVF과정에서 185개의 여포를 얻어 여포액내 GTH의 생물학적 또는 면역학적 활성을 측정하였다. Bioactive LH(bLH)는 생쥐의 Leydig cell-testosterone production assay, bFSH는 흰쥐의 Sertoli cell aromatase assay로 측정 하였 다. Immunological GTH(iLH , iFSH) 는 MaiaClone RIA , Delfia kits를 사용하였다. 여포액내 iLH, iFSH , ihCG 는 hyperstimulation에 의해 형성된 여포의 크기와는 무관하였다. 또 hMG, huFSH 의 처리와도 상관성이 없었다. T의 농도가 높은 여포액내의 iFSH는 현저히 낮았으며 E, P 가 고농도인 여포의 ihCG 양은 현저히 낮았다. 과배란이 유도된 난소의 여포액내 iLH는 LH specific RIA로 측정시 3mIU/ml 이하이었다. 생식주기중 여포액내 bLH, bFSH는 배란기에 현저히 증가 하였다. 혈청내 GTH B/I ratio는 엘정한 반면 여포액내 LH,FSH의 생물학적, 면역학적 활성은 미수정란을 가지거나 폐쇄된 여포내의 활성보다 현저하게 높았다. 위의 결과로 보아 여포액내 생식소자극호르온은 면역학적활성보다 높은 생물학적 활성을 가지며, 생리적 현상의 지표가 된다고 추론된다. 또한 steroid, bGTH는 여포의 선택, 폐쇄를 구분하는 지표로 사용가능하며, 여포가 폐쇄될때 여포액내 B/I ratio가 현저히 낮아지는 것으로 보아 GTH의 활성이 감소되는 것으로 판단된다.

면역조직화학법을 이용한 점농어 (Lateolabrax sp.) 뇌에서 두 종류 (sGnRH, cGnRH-II) 의 생식소자극호르몬 분비호르몬의 동정

김정우,이원교,양석우,정관식,조용철,노용길,방인철,김광수,임상구,유명식,권혁방,KIM Jung-Woo,LEE Won-Kyo,YANG Seok-Woo,JEONG Kwan-Sik,CHO Yong-Chul,RHO Yong-Gil,BANG In-Chul,KIM Kwang-Soo,KIM Sang-Koo,YOO Myung-Sik,KWON Hyuk-Bang 한국수산과학회 1999 한국수산과학회지 Vol.32 No.3

Two forms of gonadotropin releasing hormone (GnRH) are identified in the brain of adult mature spotted sea bass (Lateolabrax sp.) by immunohistochemical methods. Salmon GnRH immunoreactive (sGnRH-ir) cell bodies were distributed in the olfactory bulb, ventral telencephalon and preoptic region. Immunoreactive fibers were observed in the vicinity of the brain including the olfactory bulbs, the telencephalon, the optic nerve, the optic tectum, the cerebellum, the medulla oblongata and rostral spinal cord. In most cases, these fibers did not form well defined bundles. However, there was a clear continuum of immunoreactive fibers, extending from the olfactory bulbs to the pituitary. cGnRH-II-ir cell bodies were only found in olfactory bulbs. However, the distribution of cGnRH-II-ir fibers was basically similar to that of sGnRH-ir fibers except for the absence of their continuity between the olfactory bulbs and the pituitary. These data suggest that sGnRH and cGnRH-II are endogenous peptides and indicate the presence of multiple neuroendocrine functions in the brain of the spotted sea bass. It seems that sGnRH not only regulates GTH secretion but also functions as a neurotransmitter, whereas cGnRH-II functions only as a neurotransmitter. 성숙 점농어 뇌에서 세 종류의 생식소자극호르몬 분비호르몬 (GnRH)의 소재를 면역조직화학법에 의해 동정하였다. sGnRH 양성 신경세포체는 후각망울, 복측 종뇌와 전시각 지역에 분포하였다. 양성 신경섬유는 후각망울에서부터 척수에 이르기까지 다양하게 분포하였다. 면역신경섬유는 뇌의 전지역인 후각망울, 종뇌, 시각시개, 소뇌, 연수 그리고 머리쪽 척수에서 발견되었다. 대부분의 경우 이들은 모두 다발을 형성하지는 않았다. 그러나 후각망울에서 뇌하수체로 뻗어있는 양성 신경섬유는 가장 뚜렷하였다. cGnRH-II 양성 신경세포체는 후엽에서 발견되었다. 그러나 cGnRH-II 면역신경섬유도 후각망울에서 뇌하수체로 뻗은 면역신경섬유를 제외하고는 기본적으로 sGnRH 양성 신경섬유와 분포가 유사했다. 이것은 점농어 뇌에서 sGnRH와 cGnRH-II가 알려진 내인성 펩타이드이며, 이들이 다양한 신경내분비 기능을 수행할 것이라는 점을 의미한다. sGnRH는 GTH의 분비를 조절 할 뿐만 아니라 신경전달조절자로서, cGnRH-II는 단지 신경전달조절자로서 작용할 것으로 생각된다.

생쥐 초기배아에 미치는 스테로이드 홀-몬의 영향

권혁방,정순오 최신의학사 1976 最新醫學 Vol.19 No.2

The experiments were undertaken in order to study the effect of various steroid hormones on germinal vesicle break-down of mouse oocytes and to determine if there was a different sensitivity of various aged preimplantation mouse embryos to progesterone. Follicular oocytes were obtained from ovaries of A-strain female mouse and early embryos were flushed from their fallopian tubes after an appropriate time of mating. The incubation medium was a modified Krebs-Ringer bicarbonate sol. supplemented with 0.4% bovine serum alumin (BSA). A microtube culture method (Cho, 1974) for the oocytes and embryos were adopted to avoid contamination of steroid hormones which are soluble in oil. Ten to twenty oocytes or embryos were placed in the culture medium in microtube and cultured at 37°C, gassed with 5% CO2 in air humidified in an incubator. After culture, the oocytes or embryos were observed directly through the microtube under an inverted phase microscope. The results from the experiments were as follows. 1. The activity of the steroid hormones on germinal vesicle break-kown of mouse oocytes was dependent upon specificity of the hormones. 2. Oocyte was very resistant to progesterone in germinal vesicle break-down whereas fertilized ova was very sensitive to that by blocking the development of the embryos, from 1-cell to 2-cell stage. 3. When 1-cell, 2-cell, 8-16 cell embryos were cultured for 24 hours in medium containing progesterone, 8-16 cell embryos were more resistant to progesterone than the others in cleavage inhibition. 4. When the embryos were exposed to progesterone long, they became more sensitive to the hormone.