점망둑(Chasmichthys dolichognathus)의 성숙기 난모세포에서의 $C_{21}$-스테로이드 생성

백혜자,Baek, Hea-Ja 한국발생생물학회 2008 발생과 생식 Vol.12 No.1

To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

범가자미 , Verasper variegatus 의 생식소 발달단계에 따른 혈중 난황단백전구체 ( vitellogenin ) 와 성 스테로이드 호르몬 변화

백혜자(Hea Ja Baek),한창희(Chang Hee Han),김윤(Yoon Kim),(Katsumi Aida),(Makito Kobayashi) 한국수산과학회 1999 한국수산과학회지 Vol.32 No.5

Annual plasma levels of vitellogenin and sex steroids were investigated in relation to the gonadal development for understanding the endocrine control of reproduction in spotted flounder, Verasper variegatus. The plasma vitellogenin level was highest, 6.36 ㎎/㎖, in November when vitellogenesis was most active. The level, thereafter, decreased to 3.81 ㎎/㎖ in December with the initiation of spawning. On the other hand, estradiol-17β was highest, 2.7 ng/㎖, in December, and rapidly decreased in January when spawning occurred. The decreased level of estradiol-17β, around 0.2 ng/㎖, remained unchanged until May. The profiles of plasma testosterone were similar to those of estradiol-17β in the fish. The plasma 17 α-hydroxyprogesterone level was relatively low throughout the spawning period, but increased slightly with the initiation of ovarian development. In males, the plasma testosterone and 11-ketotestosterone were highest in December when spermiation actively proceeded, but rapidly decreased during the spawning period (January).

점농어 , Lateolabrax maculatus 난소에서 생성되는 C21 - 스테로이드

백혜자(Hea Ja Baek),안철민(Cheul Min An),김형배(Hyung Bae Kim) 한국수산과학회 2001 한국수산과학회지 Vol.34 No.6

점농어의 난소성숙과 배란과정에서 생성되는 성 스테로이드 호르몬 특히, C_(21)-스테로이드를 분석하고자 전구물질 ³H-pregneolone와 ³H-17αhydroxyprgesterone를 난소조직 배양초기에 첨가하여 24시간 배양 뒤 생성된 물질은 성숙기 난소 (난경 700∼800㎛)의 경우 corticosteroids계의 호르몬과 17α20βOHP가 관찰되었으나 17α20β 0HP만이 HPLC상에서 확인되었다. 특히 배란중인 점농어에서는 17α20βOHP, 17α20β21p 그리고 corticosterone의 존재가 뚜렷하였다. 따라서 점농어에서는 난소의 성숙과 배란과정에 17α20βOHP이 모두 관여하며, 배란에 이르면서 17α20β21P의 출현이 뚜렷한 것으로 생각된다. corticosterone의 존재에 대해서는 이 물질이 점농어의 배란과 관련이 있는지 또는 스트레스 조건에서 나타나는 순간적인 생성물인지 앞으로 더욱 상세한 분석방법으로 접근해야 할 부분이다. To investigate the production of C_(21)-steroids during the spawning period of spotted sea bass, Lateolabrax maculatus, we have incubated maturing and ovulating follicles with radiolabeled pregnenolone and 17α-hydroxyprogesterone for 24 hours. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). When maturing follicles (700∼800㎛ in diameters) were incubated with radiolabeled precursors, C_(21)-metabolites were corticosteroids and 17α-hydroxy,20β-dihydroprogesterone (17α20βOHP). When ovulation follicles (1,000∼1,150 ㎛ in diameters) were incubated with radiolabeled precursors, the major C_(21)-metabolites were 17α20βOHP, 17α20β,21-trihydroxy-4-pregnen-3-one (17α20β21P), and corticosterone. Additional chromatography by TLC and HPLC confirmed the presence of radioactive 17α20βOHP in the maturing follicles, and 17α20βOHP, 17α20β21P and corticosterone in ovulating follicles. Although 17α20βOHP was found in a small peak, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process. Whereas ovulation is regulated by both 17α20βOHP and 17α20β21P in the spotted sea bass. In addition, an unusual finding was the biosynthesis of corticosterone. Whether this production is responsible for the ovulation, and is an area requiring continued research.

넙치 ( Paralichthys olivaceus ) 의 in vitro 난모세포 성숙과 배란에 미치는 TBT , TPhT 및 Aroclor 1254 의 영향

백혜자(Hea Ja Baek),정지현(Jee Hyun Jung),전중균(Joong Kyun Jeon) 한국수산과학회 2001 한국수산과학회지 Vol.34 No.6

성숙 시기의 넙치 (Paralichthys olivaceus) 어미를 대상으로 난모세포의 GVBD와 배란유도과정에 TBT/TPhT와 Aroclor 1254의 저해효과를 조사한 결과, TBT (또는 TPhT)와 Aroclor 1254 모두 GVBD 과정과 HCG로 전처리한 배란유도 과정에 저해효과를 보였으며, 난모세포의 반응은 Aroclor 1254보다는 TBT에 좀 더 민감한 것으로 나타났다. TBT의 처리 농도별 차이는 TBT 0.1과 1 ppm에서 가장 낮은 GVBD 유도율을 보였으나 0.0001∼1 ppm 사이에 유의한 차이는 관찰할 수 없었다. TBT의 배란저해 효과는 HCG 처리구에 비해 HCG+TBT의 모든 실험구 (0.01, 0.1, 1 ppm)에서 뚜렷하게 나타났으며, TBT 처리구 중 가장 높은 농도인 HCG+TBT 1 ppm에서 가장 낮은 배란율을 보였다. 넙치의 GVBD와 배란과정을 유기주석화합물 (TBT, TPhT)이 방해함으로써 이 시기의 주요 호르몬인 progestogens 작용이 저해 되는 것으로 생각되며, 앞으로 난소발달 단계별로 세분화하여 그 저해 작용 메커니즘에 대한 실험이 요구된다. The effects of tributyltin (TBT), triphenyltin (TPhT) and Aroclor 1254 on germinal vesicle breakdown (GVBD) and ovulation of olive flounder (Paralichthys olivaceus) were investigated in in vitro bioassay. TBT, TPhT and Aroclor 1254 showed the inhibition effects on GVBD and ovulation in response to HCG. The oocyte response appeared to be more sensitive to TBT than Aroclor 1254. TBT was more effective in inhibiting GVBD at concentrations of 0.1 and 1 ppm. However, no significant inhibition was observed in concentrations tested (0.0001∼1 ppm). Significant inhibition of ovulation in response to HCG occurred at TBT (0.01, 0.1, 1 ppm), TPhT (0.01, 0.1, 1 ppm) and Aroclor 1254 (0.01, 1 ppm, except 0.1 ppm), compared to HCG control. The lowest ovulation rate was measured at 1 ppm TBT. These data suggest that TBT (or TPhT) could possibly interfere the actions of progestogens to induce GVBD and ovulation in in vitro bioassay system.

범가자미, Verasper variegatus의 난모세포 성숙(GVBD) 유도를 위한 HCG와 스테로이드 호르몬의 in vitro 효과

백혜자(Hea-Ja Baek),김윤(Yoon Kim) 한국수산과학회 양식분과 1996 韓國養殖學會誌 Vol.9 No.1

호르몬 처리에 의한 범가자미, Verasper variegatus의 성숙과 배란유도

백혜자(Hea Ja Baek),김윤(Yoon Kim),안철민(Cheul Min An),조기채(Kee Chae Cho),명정인(Jeong In Myeong),이남주(Nam Ju Lee) 한국수산과학회 양식분과 2000 韓國養殖學會誌 Vol.13 No.1

Bisphenol A가 점망둑 (Chasmichthys dolichognathus)의 난소 스테로이드 호르몬 대사에 미치는 영향

백혜자 ( Hea Ja Baek ),박명희 ( Myoung Hee Park ),이영돈 ( Young Don Lee ),김형배 ( Hyung Bae Kim ),김재원 ( Jae Won Kim ),유명숙 ( Myoung Suk Yoo ) 한국수산과학회 2004 한국수산과학회지 Vol.37 No.3

N/A The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor ³H-17α hydroxyprogesterone (³H-17αOHP). Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens [17α-hydroxy,20adihydroprogesterone (17α20αOHP) and 17α-hydroxy,20β-dihydroprogesterone (17α20βOHP)], androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone (E₁) and estradiol-17β (E₂)]. BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of ³H-17αOHP to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.

무지개송어(Oncorhynchus mykiss)의 난소내 협막층(theca Layers)에서의 pregnenolone 대사: cyanoketone과 trilostane의 저해 효과

백혜자 ( Hea Ja Baek ),( Alex Fostier ) 한국수산학회 1995 한국수산과학회지 Vol.28 No.4

3β히드록시-△5-스테로이드 탈수소효소(3β-HSD, 3β-hydroxy-△5-steroid dehydrogenase: △5-스테로이드→△4-스테로이드로의 대사경로에 관여하는 효소)에 대하여 특이적 저해제인 cyanoketone과 trilostane의 저해 효과가 3H-pregnenolone 전구체를 이용하여 무지개송어 난소에서 분리한 여포층, 협막층(theca layers)을 대상으로 비교 관찰되었다. Pregnenolone으로부터 17α-hydroxyprogesterone으로의 대사과정에서 주요 효소인 3β-HSD 활성은cyanoketone 10(-6)과 10(-5) M, 그리고 trilostane 10(-5)과 10(-4) M의 농도에서 억제되었으며, trilostane이 cyanoketone보다 더 효과적인 억제반응을 보이는것으로 나타났다. Pregnenolone으로부터 △4-스테로이드 대사물 축적에 대한 trilostane의 저해작용은 사용한 농도 즉 10(-8), 10(-7), 10(-6) 그리고 10-5 M에 비례하여 나타났으나 완전한 저해효과는 보이지 않았다. The effects of specific inhibitors of 3β-hydroxy-△5-steroid dehydrogenase (3β-HSD; an enzyme catalyzing conversion of △5 steroids to △4 steroids), cyanoketone and trilostane, on 3H-pregnenolone metabolism in isolated ovarian thecal layers have been investigated in vitro. At all doses of cyanoketone (10(-6) and 10(-5) M) and trilostane (10(-5) and 10(-4) M), 3β-HSD enzyme activity that transforms pregnenolone to 17α-hydroxyprogesterone was inhibited in the thecal layers. Trilostane appeared to be more efficient than cyanoketone. Trilostane at doses of 10(-8), 10(-7), 10(-6) and 10(-5) M/ml caused a doseresponse inhibition of △4 steroids accumulation in the medium from pregnenolone, but not completely blocked the conversion of △5 to △4 steroids.

한국 동해 해역에서 채집된 살오징어 (Todarodes pacificus)의 성성숙과 생식주기

백혜자 ( Hea Ja Baek ),김재원 ( Jae Won Kim ),조윤정 ( Yun Jung Cho ),김수암 ( Su Am Kim ) 한국수산과학회 2006 한국수산과학회지 Vol.39 No.6

문치가자미 (Limanda yokohamae)피부 상피층의 미세구조

이정식,강주찬,백혜자,Lee, Jung-Sick,Kang, Ju-Chan,Baek, Hea-Ja 한국현미경학회 2000 Applied microscopy Vol.30 No.3

문치가자미의 피부 상피층은 지지세포, 선세포 그리고 부속세포들로 구성된다. 지지세포는 표면세포, 중간세포 및 기저세포로 구분된다. 지지세포들의 세포질은 공통적으로 피질부와 수질부로 나누어지는때, 수질부에는 세포 소기관의 발달이 현저하며, 피질부에는 미세섬유의 발달이 뚜렷하다. 선세포들은 상피의 표면층과 중간층에 존재한다. 점액세포의 세포질은 AB-PAS에 청색으로 반응하였다. 곤봉방세포는 세포질에 원형의 중심공포와 발달된 미세섬유들을 가진다. 과립세포는 주로 중간층과 기저층에 존재하고, 세포질의 대부분은 막을 가진 전자밀도가 높은 과립들이 차지한다. 염세포는 주로 표층에 위치하며, 세포질의 대부분은 잘 발달된 미토콘드리아들이 차지한다. 색소세포는 세포질에 존재하는 함유물의 전자밀도에 따라 세 종류로 구분할 수 있다. Ultrastructures on the integumentary epidermis of the marbled sole, Limanda yokahamae, were examined by means of the light and transmission electron microscope. Epidermal layer consists of supporting cells, unicellular glands and accessory cells. The supporting cells were classified into superficial cell, intermediated cell and basal cell. The cytoplasm of supporting cells is divided into cortex and medullar part. In the cortex and medullar part, microfilaments and cell organelles are well developed, respectively. Gland cells are present in the superficial and middle epidermis. The cytoplasm of mucous cell reacted to blue in AB-PAS (pH 2.5). Club cell has a roundish central vacuole and well-developed microfilaments in the cytoplasm. Granular cells are occurs in the middle and basal epidermis , and the cytoplasm is occupied with membrane-bounded granules of electron dense. Chloride cells are present in the superficial epidermis , and the cytoplasm is occupied with tubular mitochondria. Three types of pigment cells can be distinguished by electron density of cytoplasmic inclusions.