• Title/Summary/Keyword: 과립 분비

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한우 난소 과립막세포의 체외배양에서 Follicular Fluid 및 Anti-Inhibin Serum의 첨가효과

  • 성환후;최선호;장유민;박진기;장원경;정일정
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.33-33
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    • 2001
  • 본 연구는 한우 난포발달에 있어서 난포액 및 inhibin의 생리적 역할을 검토하기 위해 수행하였다. Anti-inhibin serum(AIS) 생산을 위해 사용된 항원은 porcine inhibin-$\alpha$-subunit 19~32의 peptide를 사용하여 adjuvant 용액을 1:3의 비율로 혼합하여 앙고라종 토끼 5두(체중 2.5kg)에게 주 2회 간격으로 8회 실시후 ELISA Leader로 항혈청의 역가를 확인하였다. 난포액(bFF; bovine follicular fluid)은 도축장에서 도축되는 한우 난소로부터 직경 1.0cm 이하의 난포로부터 회수하여 스테로이드를 제거하기 위해 10% chacoal solution(50 mg/$m\ell$, Norrit-A, Fisher Sci., USA)을 처리하여 45분간 배양후 원심분리후 상층액을 회수하여 실험에 공시하였다. 과립막세포의 체외배양을 위해 D-MEM용액(10% FCS와 antibiotics를 첨가)을 배양액으로 하여 1 $\times$ $10^{6}$ cells/$m\ell$로 조절하였다. 호르몬은 RIA 및 ELISA법으로 분석하였다. 그 결과 항원-항체반응은 항원처리후 24일째부터 항체가를 확인할 수 있었으며 52일째에서 높은 항원-항체반응을 보였다. 한편, 난포크기별 난포액의 progesterone 및 Estradiol-l7$\beta$을 농도를 분석한 결과, Progesterone 난포크기가 직경 1.0 cm부터 유의적으로 증가하기 시작하여 직경 2.0cm의 난포액에서는 높은 progesterone이 존재하고 있는 것으로 나타났다. 이에 반해, 난포크기별 난포액중 estradiol 17$\beta$농도는 직경 1.0cm구에서 가장 높게 나타났다. 난포직경이 1cm일 경우에 난포액내 etradiol-17$\beta$가 가장 많이 존재하고 있음이 나타났다. 과립막세포의 체외배양에서 배양 24시간에서는 과립막세포의 progesterone분비는 약 40ng/1$\times$$10^{6}$ cell/well/$m\ell$ 전후로 나타났으며 bFF 5%, AI 5% 및 bFF+AI 첨가구에 따라 유의적인 차이는 없었다 반면에 48시간배양구에서는 24시간에 비해 유의적으로 높게 분비되었으며, bFF 5%처리구와 bFF5%+AI5%처리구에서는 progesterone을 대조구보다 유의적으로 억제되었으나 AI 5%단독처리구에서는 대조구와 큰 차이가 없었다. 한편, 각 세포질을 SDS-PAGE로 분리하여 nitro cellulose membrane에 transfer하여 Western blotting법에 의해 검토한 결과, 직경 1.0 cm의 성숙난포의 granulosa cell에 특이하게 Inhibin이 존재하고 있음이 확인되었다. 이상의 결과로, 한우에 있어서 성숙난포에 존재하는 Inhibin은 난포발달 및 난포세포의 스테로이드호르몬합성에 중요하게 관여하고 있는 것으로 사료된다.

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Change of Steroid Receptor Number of and Bioactivity of Gonadotropins in the Follicular Fluid of Porcine Ovarian Atretic Follicles(II):Testosterone Receptor (돼지 폐쇄여포내 스테로이드 수용수체의 변화와 여포액내 생식소 자극호르몬의 활성도 변화(II): 테스토스테론 수용체)

  • Yoon, Yong-Dal;Lee, Chang-Joo;Chun, Eun-Hyun;Kim, Moon-Kyoo;Lee, Joon-Yeong
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.1
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    • pp.23-34
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    • 1989
  • Estradiol (E)은 난소내 과립세포 (granulosa cell, GC)를 증가시키고 생식소 자극호르몬과 협동으로 배란을 유도한다. Androgen은 E의 작용과 반대의 작용을 나타내며 여포의 폐쇄요인으로 알려지고 있다. Testosterone(T)이 폐쇄여포의 여포액내 다량 존재하는 것이 알려짐에 따라 난소내에도 자가조절분비(autocrine)또는 paracrine regulation에 의해 작용을 나타낼 것으로 가정되어 난소내 여포가 폐쇄됨에 따라 그 수용체의 변화하는 양상을 조사하고져 하였다. 흰쥐의 과립세포의 세포질에는 $51.3{\pm}6.1$fmol/mg protein의 Estradiol 수용체(ER) ; $153.1{\pm}25.3$의 Testosterone수용체(TR) ; 또한 $35.1{\pm}8.1$의 Progesterone수용체(PR)가 존재하였다. 과립세포내 ER용 세포질내 E를 제거한 후에 정량이 가능하였고 또한 과립세포내에도 TR이 사람에서는 $23.4{\pm}7.2$ fmol/mg protein, 돼지는 $98.5{\pm}23.1$로 상당량 존재함을 관찰하였다. Dihydrotestosterone Enanthate(DHTE)를 100ug/흰쥐의 농도로 처리한 결과 난소내 TR의 농도는 변화가 없이 ER의 농도만 현저히 저하되고 쥐의 난소무게 역시 감소하는 것을 발견하였다. 위의 결과로 보아 난소내에도 스테로이드 호르몬은 autocrine(자가조절)방법으로 작용하며 An-drogen이 난소의 무게를 감소시키는 것은 ER의 수를 감소시켜 E의 작용이 억제되고 여포들이 폐쇄를 일으켜 그 증식이 저하된 때문으로 사료된다.

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Inhibitory Effects of Soyasaponins on Antigen-induced Degranulation in RBL-2H3 Cells (Soyasaponin의 RBL-2H3 세포에서 탈과립화 억제 효과)

  • Yang, Seung Hwan;Lee, Jung A;Lee, Jae Yeon;Ahn, Eun-Kyung;Shin, Tai-Sun;Tsukamoto, Chigen;Chung, Gyuhwa;Suh, Joo-Won
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.287-290
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    • 2015
  • Soyasaponins are glycosylated, which gives rise to a wide diversity of structures and functions. We evaluated for inhibitory effects of 4 soyasaponins on antigen-induced degranulation in RBL-2H3 Cells. 4 soyasaponins had shown dose-dependently inhibited histamine and ${\beta}$-hexosaminidase.

Control Mechanisms of Ovulation by Pituitary Adenylate Cyclase-Activating Polypeptide (Pituitary Adenylate Cyclase-Activating Polypeptide에 의한 배란 조절에 관한 연구)

  • Lee, Yu-Il;Kim, Hyoung-Choon;Kim, Mi-Young;Chun, Sang-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.101-111
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    • 2005
  • 배 경: Pituitary adenylate cyclase-activating polypeptide (PACAP)은 양의 시상하부에서 추출된 신경펩타이드 호르몬으로 난소에도 존재하여 배양된 과립막 세포에서 스테로이드합성과 cyclic AMP 형성을 촉진함이 보고되었다. 목 적: 흰쥐 난소를 실험 모델로 사용하여 배란시 황체화호르몬 (luteinizing hormone; LH)에 의해 유도된 PACAP과 PACAP 수용체의 유전자 발현양상과 신호 전달경로를 규명하고자 하였다. 재료 및 방법: 미성숙 흰쥐의 배란전 난포를 체외 배양하면서 LH로 처리하고 PACAP 및 PACAP수용체의 유전자 발현을 보기 위해서는 Northern blot 분석과 in situ hybridization (ISH)을, 그리고 단백질 수준의 PACAP 검색을 위해서는 enzyme linked immunosorbent assay (ELISA) 분석을 이용하였다. 결 과: LH 처리 후 Northern blot상의 PACAP 유전자 발현은 6~9시간에 일시적으로 최고치에 도달하였으며 ISH로 보아 과립막 세포에서 발현됨을 알 수 있었다. ELISA 분석 상 PACAP 단백질도 LH처리 후 6~12시간에 최고치를 나타내었으며, PACAP 수용체 mRNA 역시 3~9시간에 최고치로 과립막 세포에서 발현되었다. Adenylate cyclase (AC) 억제제인 MDL12330A 처리시 LH로 발현된 PACAP mRNA가 감소되며, AC의 활성제인 forskolin 처리에는 LH시와 유사한 PACAP mRNA의 발현양상을 나타내었다. 그러나 protein kinase C (PKC)의 억제제인 chelerythrine과 2-0-tetradecanolphorbol-13-acetate (TPA) 처리로는 PACAP 의 유전자 발현에 영향을 주지 못하였다. 5-lipoxygenase의 억제제인 MK886이나 nordihydroguaiaretic acid (NDGA)로 처리한 결과 LH로 유도된 PACAP 유전자의 발현이 감소되었으나, cyclooxygenase의 억제제인 indomethacin은 별로 영향을 주지 못하였다. MEK와 p38의 억제제인 PD98059와 SB203580도 LH로 촉진 된 PACAP의 유전자 발현을 농도 의존적으로 억제하였다. 결 론 : 배란전 난포에서 PACAP과 PACAP 수용체의 유전자 발현은 모두 LH의 폭발적 분비에 의해 유도되어 일시적으로 과립막 세포에서 나타나 배란을 위한 국소적인 조절 작용을 할 것으로 추정되며, LH로 촉진된 PACAP 유전자 발현을 위한 신호전달은 cAMP-PKA, lipoxygenase 및 MAP kinase 경로를 통하는 것으로 사료된다.

Ultrastructure of the Cutaneous Mucous Glands and Histochemical Pronerty of the Mucous Secretory Material in Bombina orientalis. (무당개구리(Bombina orientalis Boulenger) 피부 점액선의 세포구조 및 점액분비물질의 조직화학적 특성에 관한 연구)

  • 문명진;김기영;김우갑
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.221-231
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    • 1989
  • The histochemical characteristics of the cutaneous mucous glands and its fine structure of the Korean fire bellied toad, Bombina orientolis Boulenger, were studied with light and electron microscopes. The mucous glands of the toad are simple alveolar gland, and are composed of inner glandular epithelial cells and outermost myoepithelial cells. Histochemical studies suggest that main secretory materials released from the mucous glands might be acid mucopolysaccharides which is more abundant in the ventral skin than in the dorsal skin. The scanning electron microscopic observation showed that the morphology of the opening sites of the gland duct is similar to the morphology of stomata on the plant leaves. Glandular epithelial layers of the mucous glands are composed of two cell types, the mucous secreting cells and the mitochondria rich cells. The mucous secreting cells could be classified according to the morphology and inner textures of cytoplasm and those characteristics were seemed to be related to the degree of maturation of the cells. Furthermore the mucous within the mucous secreting cells appears to be synthesized in the rough endoplasmic reticulum.

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Immuno-Electronmicroscopic Studies on the Expression of Tenascin in the Synovial Cells of Human Knee Joint (퇴행성관절염의 사람 무릎관절 윤활막에서 Tenascin 발현에 대한 면역전자현미경적 연구)

  • Lim, Hyoung-Soo;Choi, Hee-Joon;Lee, Se-Jeong;Hwang, Douk-Ho
    • Applied Microscopy
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    • v.35 no.2
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    • pp.73-80
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    • 2005
  • To observe the morphological changes and the expression of tenascin in synovium of human knee joint, between normal condition and degenerative arthritis, were processed by immunoelectron microscopic method. The results were summarized as follows. 1. In degenerative arthritis, the hyperplasia of synovial membrane was characterized by the increase of cell number in secretory synovial cells. 2. In normal condition, there was no marking of the immuno-gold for tenascin in synovial membrane. 3. In degenerative arthritis, the immuno-gold for tenascin were observed in endoplasmic reticulum of secretory synovial cells and extracellular matrix of synovial layer. On the basis of above findings, the hyperplasia of synovial membrane and the pathologic processes may be concerned with the increase of number of secretory synovial cells and of expression of tenascin, in degenerative arthritis.

A Histological Study of Skin on Some Amphibia Inhabitated Chiri Mt. and Moodeung Mt. (지리산과 무등산에 서식하는 한국산 양서류의 피부에 관한 연구)

  • 이승휘;권은호;신영희
    • Korean Journal of Environment and Ecology
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    • v.18 no.2
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    • pp.205-209
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    • 2004
  • For the purpose of this study was accumulating histological data of skin some amphibia near Chiri Mt. and Moodeung Mt. Analyzed Anura and Caudata were Rana nigromaculata, Rana rugosa, Rana catesbeiana, Hynobius leechii. The histological prepared skin of frogs were compared, of which were selected from dorsal and belly. Excretory glands were identified granular glands, mucous glands, serous glands, vacuoles and excretory ducts in epidermal and dermal tissue. And developing excretory glands, well developed excretory glands and post developing excretory glands were identified also. These results were significantly as basal data on the comparative epidermal skin histology on some Korean Amphibia. Probably these glands of amphibian skin could be infered which were adaptable structure to ecological suffered condition. Following study of these results were more considerable data for comparative histology, comparative anatomy and comparative physiology and ecology of Amphibia.

Integumentary Ultrastructure of the Blenny, Pholis nebulosa (Teleostei: Pholidae) (베도라치 (Pholis nebulosa) 피부계의 미세구조)

  • LEE Jung Sick;An Cheul Min;HUH Sung-Hoi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.2
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    • pp.148-152
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    • 2000
  • To provide basic information on the integumentary system of the blenny, Pholis nebulosa, ultrastructures of epidermal and dermal layer were examined by means of the light and transmission electron microscope. The skin of the blenny consisted of epidermal and dermal layer. Epidermal layer consisted of supporting cell and unicellular gland. The supporting cells were classified into superficial cell, intermediated cell and basal cell, and the gland cells were classified into mucous secretory cell and club cell. The cytoplasm of supporting cells was divided into cortex and medullar part. In the cortex and medullar part, microfilaments and cell organelles were well developed, respectively. Superficial cell of epidermal layer was cuboidal and contained nucleus of horseshoe shape. Intermediated cell had a nucleus of irregular form and the electron density was higher than the other supporting cells, Basal cell was columnar, but nucleus was situated in the upper cytoplasm. Cell organelles of the basal cell were poor than the other supporting cells, but membrane interdigitations were well developed. The cytoplasm of mucous secretory cell had a well-developed ovoid secretory granules, which reacted to red with AB-PAS reaction. The club cell had a we31-developed round secretory granules and endoplasmic reticulum. figment cells were classified into two type. The one contained pigment granules of electron dense, and the other contained reflecting platelets. The cytoplasm of fibrocyte had n well developed rough endoplasmic reticulum.

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Cellular Immune Response of Protaetia brevitarsis seulensis Larvae to Metabolites Produced by Pathogenic and Symbiotic Bacteria (흰점박이꽃무지(Protaetia brevitarsis seulensis) 유충에서 병원균과 공생균 분비물질들에 의한 세포성면역반응)

  • Hwang, Dooseon;Cho, Saeyoull
    • Korean journal of applied entomology
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    • v.57 no.1
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    • pp.25-32
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    • 2018
  • Studies of immune responses in insects have focused on mechanisms that interact directly with invading microorganisms. However, few studies have examined the immune response to various metabolites produced by microorganisms after they enter the host. Here, we examined immune responses in Protaetia brevitarsis seulensis larvae induced by metabolites produced by symbiotic and pathogenic bacteria. The two types of bacteria were cultured under the same conditions. The bacteria were then removed and the remaining culture supernatant was injected into the larvae. The larvae injected with culture medium (Ch-medium) from symbiotic bacteria remained relatively healthy and did not develop an immune response, whereas more than 60% of the larvae injected with pathogen culture medium (Ec-medium) died after 150 hours and dark brown patches of melanin were observed at the injection site. This immune response was confirmed by the finding of activated lysosomes in insect granulocytes. More than 50% of lysosomes in larvae injected with pathogen culture medium were strongly stained after 12 h, but less than 5% of those injected with symbiotic culture media were stained. Therefore, it is assumed that symbiotic bacteria produce few (if any) substances that induce host immune responses.

Ultrastructure of the Androgenic Gland of the Freshwater Prawn, Macrobrachium nipponense (징거미새우, Macrobrachium nipponense의 Androgenic Gland 미세구조)

  • 김대현;강정하;김대중;한창희
    • Development and Reproduction
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    • v.3 no.1
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    • pp.53-58
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    • 1999
  • The androgenic gland secretes a hormone, androgenic gland hormone, which is believed to act on the differentiation of the primary, secondary, and behavioral sex characteristics in most malacostracan crustaceans. This report presents the ultrastructural morphology of the androgenic gland in the freshwater prawn Macrobrachium nipponense. This gland, located in the coxopodite of the last pair of walking legs, was attached to the subterminal region of the sperm duct. The gland was composed of simple cellular strands, encased by a fibrous sheath. Microvilli were situated in the fibrous sheath, especially at the edge of each cellular strand. The androgenic gland cells had the large and round nucleus and rough endoplasmic reticulum arranged either in spirals or in concentric circles throughout the cytoplasm of the cell. They also had the well-developed Golgi complex and long mitochondria with flat and transverse cristae. The Golgi complex was similar to microvesicular cluster, but usually in the shape of typical dictyosomes, These features of androgenic gland cells coincides well with the protein/peptide secretion in their function. However, despite the apparent ultastructual equipment for protein/peptide secretion, no accumulation of materials secreted were noticed in the cytoplasm. Therefore it is strongly suggested that the transient transportation of the materials into the hemocoel has occurred just after synthesis.

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