Alteration of Acrosome Reacting Ability of Mouse Spermatozoa during Epididymal Transit

계명찬,김성례,김문규,Gye, Myung-Chan,Kim, Sung-Rye,Kim, Moon-Kyoo The Korean Society of Developmental Biology 1997 발생과 생식 Vol.13 No.4

생쥐의 부정소에서 진행되는 정자 성숙과정 동안의 첨체반응 능력의 변화를 조사하였다. 자발적 첨체반응 및 난포액, 프로게스테론, 또는 A23187에 의해 유발되는 첨체반응은 모두 정자의 성숙에 의존저긍로 일어났다. 두부 부정소의 매우 적은 정자만이 난포액 및 프로게스테론에 반응하여 첨체반응을 일으켰으며 체부 및 미부 부정소간에 첨체반응율에 차이가 없었다. 반면 A23187 처리시 상당수의 두부 부정소 정자가 첨체반응을 진행하였다. 이러한 결과에서 두부 부정소를 거쳐 체부 부정소에 도달한 생쥐 정자는 첨체반응 능력을 획득하여 이 과정에서 정자의 원형질막 표면에서는 첨체반응을 유발하는 물질과 상호작용에 필요한 변화가 진행되는 것으로 사료된다. 반면 정자내로의 $Ca^{2+}$ 유입 후 진행되는 막융합과 첨체내용물의 분비에 필요한 능력은 두부 부정소 정자도 일부 갖고 있는 것으로 사료된다.

북한강 지류산 꺽지 ( Coreoperca herzi ) 의 생식소 성숙에 관한 연구

계명찬,정규회,유병선 ( Myung Chan Gye,Kyu Hoi Jung,Byung Sun Yoo ) 한국환경생물학회 1997 환경생물 : 환경생물학회지 Vol.15 No.2

Gonadal maturation of Korean brook perch Coreoperca herzi was studied. Samples were collected in the streams of Pukhan River from May to October 1996. Sex ratio of samples collected was 1 : 2 (female : male). Individuals larger than 100mm in their body length started to grow germ cells in their gonads. Gonadosomatic index (GSI) ranged from 1.366 to 10.744 in females .and from 0.082 to 0.994 in males. GSI was the highest at late June in both sexes. This result suggests that Coreoperca herzi breeded in the late June in the streams flowing into Pukhan River.

저정낭액이 생쥐 부정소 정자의 첨체반응에 미치는 영향

계명찬,김성례,김문규,Gye, Myung-Chan,Kim, Sung-Rye,Kim, Moon-Kyoo 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.1

This study aimed to evaluate the effect of seminal vesicle fluid (SVF) on the acrosome reaction (AR) occurred spontaneously or induced by $Ca^{2+}$ ionophore A23187, follicular fluid, and progesterone in mouse epididymal sperm. SVF was divided into high (MW>10 kD) and low (MW<10 kD) fractions by ultrafiltration. The low MW fraction of SVF decreased the rate of spontaneous AR, however the high MW fraction did not. It suggested that the low MW fraction of SVF might have contained decapacitation factor(s) responsible for prolonging of time need for capacitation. When sperm preincubated for 60 min in the presence of SVF, the rate of AR induced by A23187 was decreased, but prolongation of preincubation time for 120 min significantly potentiated the AR by A23187. It suggested that addition of SVF into sperm preincubation medium imposed the epididymal sperm a condition similar to ejaculation. AR induced by human follicular fluid or progesterone was also inhibited by SVF. It suggested that substance in SVF might have affected AR of mouse sperm by inhibiting the interaction between AR inducing ligands and sperm surface receptors involved in acrosomal exocytosis.

꺽지 ( Coreoperca herzi ) 의 정자형성

계명찬(Myung Chan Gye) 한국하천호수학회 2002 생태와 환경 Vol.35 No.3

In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ㎛ in length. The round head was 2.2 ㎛ long. The middle piece developed beneath the head was 0.5 ㎛ long. Tail was 40 ㎛ in length. Coomassie brilliant blue(CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.

무당개구리 비텔로제닌 유전자의 발현의 RT-PCR 검출법

계명찬 ( Myung Chan Gye ),이명식 ( Myung Sik Lee ),강희정 ( Hee Jung Kang ),정경아 ( Kyung Ah Jung ),안혜선 ( Hae Sun Ahn ) 한국환경생물학회 2004 환경생물 : 환경생물학회지 Vol.22 No.2

N/A To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using 1 μg of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT-PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a 2^(-12) equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.

풀망둑 난황전구단백질 유전자발현 추적기법

계명찬 ( Myung Chan Gye ) 한국환경생물학회 2004 환경생물 : 환경생물학회지 Vol.22 No.1

N/A In an effort to develop the tools for monitoring the contamination of xenoestrogen in the aquatic environment of Korea, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (VTG) mRNA expression were optimized in Synechogobius hastus. Based on the partial VTG cDNA sequence VTG mRNA level in livers from male fishes was analyzed by RT-PCR. As an internal control beta actin mRNA was amplified. 3 μg of total RNA was reverse transcribed in 20 μl reaction using murine leukemia virus [MuLV] reverse transcriptase. Subsequent PCR using the 1 μg of cDNA resulted in linear increase in PCR product of VTG in female liver cDNA from 10 to 30 cycles of amplification. On the contrary, in male, PCR product first detected at 28 cycles of amplification and linearly increased during 38 cycles of amplification, suggesting that male S. hastus expresses minute amount of VTG mRNA which is 2^(-18) equivalent of female. In conclusion, the optimized protocol of VTG mRNA expression in the liver of male S. hastus will be promising the environmental monitoring the xenoestrogen contamination in the western coast and estuaries in Korea.

극지역 물개류의 번식생물학: 정자형성

계명찬 ( Myung Chan Gye ),강성호 ( Sung Ho Kang ) 한국환경생물학회 2006 환경생물 : 환경생물학회지 Vol.24 No.1

돼지 난포내 Alkaline Phosphatase 및 Adenosine Triphosphatase의 활성부위와 활성도

김문규,계명찬,윤현수,김종흡,Kim, Moon-Kyoo,Gye, Myung-Chan,Yoon, Hyun-Soo,Kim, Jong-Heup 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.2

In order to study the growth and maturation of ovarian follicle, the localization and activity of alkaline phosphatase(ALPase) and adenosine triphosphatase(ATPase) of the granulosa cells and theca layer were examined according to the follicle size, the follicle state and the ovarian cyclic phase in pig. Theca interna of the small follicles was more heavyly localized with reaction product by the activites of ALPase and ATPase than that of the large follicles. It is assumed that, as the follicles proceed to growth and maturation, antrum formation is the result of the follicular fluid accumulation by means of active transport by the activities of ALPase and ATPase in theca interna. The activities of ALPase and ATPase in atretic follicles were higher than those of normal follicles. These results imply that the mechanisms of follicle maturation and atresia are different according to the phase of ovarian cycle.

Indomethacin이 생쥐 착상전 배아의 발생 및 부화에 미치는 영향

전용필,계명찬,김정훈,김문규,Cheon, Yong-Pil,Gye, Myung-Chan,Kim, Chung-Hoon,Kim, Moon-Kyoo 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.1

The present study was designed to define the role of prostaglandin in the development and hatching of mouse embryo. The effects of indomethacin, an inhibitor of prostaglandin synthesis, on the development and hatching of morula and blastocyst were examined. In early morula stage, embryos were degenerated significantly at 100 ${\mu}M$ and 200 ${\mu}M$ indomethacin. However, the viability of embryos was not influenced by concentration in any other embryonic stages. In all embryonic stages, the hatching was suppressed with concentration dependent manner, but expansion was not suppressed. Particularly, in 84h embryos post hCG injection, the hatching was suppressed significantly compared with post hCG 72h or 96h embryos. When embryos were treated with 100 ${\mu}M$ indomethacin for a specific time (12h) in according to the development stage, the hatching was suppressed all groups. These suppressional effect was decreased as embryonic development stage was progressed. However, the expansion was not affected in all treatment group. This study suggests that hatching-related metabolic substances are synthesized from morula stage and intraembryonic signaling mediated prostaglandin was important for development and hatching of mouse embryo.

Azole계열 항곰팡이 물질의 양서류 독성: 총설

박찬진 ( Chan Jin Park ),박용아 ( Yong Ah Park ),옥승석 ( Seung Seok Ok ),계명찬 ( Myung Chan Gye ) 한국환경생물학회 2014 환경생물 : 환경생물학회지 Vol.32 No.1

양서류는 육상과 수상생태계를 연결하는 먹이사슬의 연결자로 진화적 생태적 지이를 갖는다. 양서류의 배아와 유생은 모체와 독립되어 수환경 내에서 초기발생 및 성장하기 때문에 수환경에 존재하는 다양한 화학물질에 직접적으로 노출될 수 있다. Azole계열 항곰팡이제는 농업 및 의료용으로 널리 사용되는 화학물질로서 농지, 하수처리장 등으로 부터 수계로 유입된다. 최근, 양서류에서 이러한 azole계 물질에 의한 기형유발, 내분비계장애 효과가 증가하고 있다. 본 소고에서는 azole계 물질의 양서류 독성을 파악하고 azole계 물질의 안전한 이용을 위한 가이드라인을 제공하고자 azole계열에 속하는 imidazole, triazole, thiazole, oxazole, pyrazole 항곰팡이 물질이 양서류의 발생, 분화, 생식 등에 미치는 영향에 대해 최근까지의 연구결과를 정리하였다. Amphibians play a pivotal role in the ecosystem as a mediator between aquatic and terrestrial environment. Currently they are directly exposed to a variety of chemicals in the aquatic environment throughout their life cycle. Azole fungicides have been widely used in medical applications and agricultural activities. The direct exposure of azole fungicides causes an alarming situation for various ecosystem. Recently, teratogenesis and endocrine disruption by azole fungicides have been reported in amphibians. In an effort to provide the current information for amphibian toxicity of azole fungicides and to make the guidelines for safe usage of azolebased materials, the effects of azole fungicides including imidazole, triazole, thiazole, oxazole, and pyrazoleon on early development, differentiation and reproduction of amphibians were reviewed.