Aquaporin 5 expression is regulated by estrogen receptor α in mouse oviductal epithelium

Won Heum Nah,Myung Chan Gye,Jung Wook Kim,Jung Hye Hwang 한국발생생물학회 2013 한국발생생물학회 학술발표대회 Vol.2013 No.8

Aquaporin5 (AQP5), a water channel plays an important role in the fluid homeostasis and cell volume control in epithelial cells. In an effort to understand fluid homeostasis in the oviduct, tissue specific expression of AQP 5 was examined together with hormonal regulation of AQP5 in the mouse oviduct. To understand the oviductal fluid homeostasis and its regulation by sex steroids, We examined AQP5 expression in mouse oviduct during developmental stage and estrous cycle, and in estrogen receptor α (ERα) knockout mice oviduct. In immature mouse oviduct, expression of AQP5 expression was examined after stimulation with gonadotropins. The effect of ERα agonist (PPT) and ERβ agonist (DPN) on the oviductal expression of AQP5 was examined in ovariectomized mouse. All samples were subjected to realtime-PCR and immunohistochemistry analysis. In oviduct epithelium, AQP5 was largely found in the apicolateral membrane and cytoplasm of ERα-positive non-ciliated cells but weakly expressed in the ciliated cells. Interstitial cells, muscle cells and blood vessels were also weakly positive for AQP5 immunoreactivity. In cyclic female mice oviductal AQP5 mRNA levels were the highest at estrous. In immature mouse oviduct AQP5 mRNA and epithelial immunoreactivity were increased by PMSG, and followed by a decrease after hCG. In ERα KO mice oviduct, AQP5 mRNA levels were significantly lower than those of WT females at diestrous stage. In immature and OVX mouse oviducts, AQP5 mRNA and epithelial immunoreactivity were significantly increased by E2 and PPT. Together, our results suggest the pivotal role of AQP5 in fluid secretion and absorption of water in non-ciliated cells in oviduct. AQP5 gene is tightly activated by estrogen – ERα signaling in non-ciliated cells in oviductal epithelium, mediating the effect of estrogen on gamete transport, fertilization and early embryo development via regulating the fluid homeostasis in oviduct.

생쥐의 oviduct에서 estrogen에 의한 bone morphogenetic protein 2, 4 (BMP2, BMP4)의 발현 조절 및 기능

조남희,계명찬 한국발생생물학회 2017 한국발생생물학회 학술발표대회 Vol.2017 No.8

골 형성 단백질(Bone morphogenetic protein, BMP)은 TGF-β superfamily의 구성원 중에 하나이며, 이들은 원래 뼈 형성을 유도하는 능력에 의해 발견되었지만, 뼈 외에도 다른 세포 및 기관의 성장과 분화를 조절하는 것으로 밝혀졌다. 다기능적인 골 형성 단백질은 포유동물의 reproduction에도 매우 중요한 역할을 하는데, 예를 들어 BMP8A와 BMP8B는 생쥐의 정자 형성과 부고환에 일정한 기능을 하는 것으로 보고되었고 난소에서의 BMP15는 난포성장을 자극하고, 과립막세포(granulosa cell)의 증식시키는데 관여하는 단백질로 알려져 있다. 하지만 난관(oviduct)에서의 BMP 역할은 알려진 바가 적다. 그래서 본 연구에서는 골 형성 단백질이 착상 전 oviductal environment에 어떠한 영향을 미치는지 밝히고자 하였다. 8주령 생쥐의 Estrous cyclic oviduct을 가지고 RT-PCR과 Immunohistochemistry(IHC)를 통해 BMP2, 4의 mRNA와 단백질 발현을 확인하였다. Estrogen에 의한 BMP2, 4의 영향을 확인하고자 난소절제술을 시행한 생쥐와 Estrogen receptor alpha(ERα) Knockout 생쥐를 통해 mRNA와 단백질 발현을 확인하였다. Oviduct의 ciliated cell을 가지고 BMP2, 4의 기능을 밝히고자 siRNA실험을 진행 하였다. Estrous cyclic oviduct cDNA를 통해 RT-PCR한 결과, 이 중 Estrus 시기에 가장 높은 발현을 보인 BMP2, 4의 mRNA level은 Isthmus보다 Infundibulum과 Ampulla에 증가하였고, Immunohistochemistry (IHC)를 통해 BMP2와 BMP4 단백질은 난관 상피세포 중에서 ciliated cell에 발현되었다. 이를 ciliated cell marker인 β-tubulin과 함께 Immunofluorescence(IF)를 진행한 결과, 주로 β-tubulin-positive ciliated cell에서 발현됨을 확인하였다. 난소 절제술을 시행한 생쥐의 난관에서는 E2와 DPN (ERβ agonist)에 의해 BMP2, 4 mRNA 발현이 증가하고, ERαKO 난관의 경우 WT(Die)에 비해 BMP2, 4 mRNA, 단백질의 발현 모두 증가하였다. BMP2와 BMP4의 기능을 밝히고자 Ciliated cell line인 OA-6b를 가지고 siRNA 실험을 진행한 결과, Bmp2, Bmp4 siRNA 처리군에서 Ciliated cell marker인 FOXJ1의 발현이 줄어들고 Proliferation marker인 Ki67, Pcna의 발현이 낮아졌다. 이로써, Oviduct 내 BMP2와 BMP4 발현은 Estrogen과 양의 상관성이 있으며, Ciliated cell에서 Estrogen - ER β signaling을 통해 조절된다. Estrogen에 의해 유도되는 BMP2와 BMP4는 ciliated cell에서 Autocrine, Paracrine factor로 작용할 가능성이 있다. 더 나아가 Oviduct의 Infundibulum 및 Ampulla에 강하게 발현되는 BMP2, BMP4는 난자 및 배아에 autocrine, paracrine factor로 작용하여 oviductal cell proliferation을 조절하고, 수정을 위한 oviductal environment 조성에 중요한 역할을 하는 것으로 사료된다.

한국산 꿩 난관의 혈관분포에 관한 해부학적 연구

최성도,이영훈,김인식,양홍현 한국가금학회 1999 韓國家禽學會誌 Vol.26 No.1

The distribution of oviductal vessels of eight Korean native pheasants was observed after Latex casts of vessels. Latex was injected into thoracic aorta and posterior vena cava of these birds for cast preparation. The results were as follows: 1. The arteries suppling oviduct of the Korean native pheasants contained A. oviductalis cranialis, media, caudalis and vaginalis. The veins drainaging oviduct were stretched along the following ateries and all of the oviductal vessels were situated on the left side of the body. 2. The anterior oviductal artery arose from the pubic artery of the left external iliac artery and distributed to the infundibulum and the magnum of the oviduct. 3. The middle oviductal artery arose from the left sciatic artery and distributed to the isthmus and shell gland of the oviduct. 4. The caudal artery arose from the left internal pudendal artery of the median sacral artery and distributed to the caudal part of uterus and the cranial part of vagina. 5. The vaginal artery arose from the left internal pudendal artery of median sacral artery and distributed to the caudal part of vagina.

Identification and Regulation of AHCYL1 Gene by Estrogen in Chickens

Wooyoung Jeong,Suzie E. Ahn,Jinyoung Kim,Fuller W. Bazer,Jae Yong Han,Gwonhwa Song 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

S-adenosylhomocysteine hydrolase-like protein 1 (AHCYL1), also known as IP3 receptor- binding protein released with IP3 (IRBIT), is a member of the AHCY-like protein family. AHCYL1 protein regulates IP3-induced Ca2+ release in the cytoplasm of cells and, therefore, is likely to be an important gene regulating various biological processes in the oviduct of chickens. Inmammals, expression is greatest during activation of dendritic cells which are antigen presenting cells associated with immunoregulatory processes in blood and skin. However, the identification of the AHCYL1 gene in chickens has not been investigated. In the present study, we first used RT-PCR to demonstrate AHCYL1 gene expression in adult chicken organs and oviducts of immature chickens treated with DES (diethylstilbesterol, a synthetic estrogen agonist). The results indicated that AHCYL1 mRNA is expressed in chicken reproductive organs (testis, ovary and oviduct). Inaddition, expression of AHCYL1 mRNA increased in response to DES-treated immature oviducts compared to the non-treated control immature oviducts of chickens. Interestingly, AHCYL1 was abundant in the cytoplasm of luminal and glandular epithelia, but not in other cell-types such as stroma and connective tissues, of the chicken oviduct. These results suggest that AHCYL1 is a novel estrogen-stimulated gene associated with development of the chicken oviduct, as well as functions of oviductal glandular and luminal epithelia that may include activation of resident immune cells, such as dendritic cells.

Differential Expression of SERPINs in the Chicken Oviduct

Whasun Lim(임화선),Jihye Kim,Suzie E. Ahn,Wooyoung Jeong,Jinyoung Kim,Jin Won Choi,Jae Yong Han,Fuller W. Bazer,Gwonhwa Song 한국가금학회 2010 한국가금학회 정기총회 및 학술발표회 Vol.27 No.-

Serpins are a superfamily of relating to inhibit protease with common structural features and inhibitory mechanisms. It is also known as ov-serpins in chicken which are ovalbumin and secreted chicken oviduct protein. However, to our knowledge, it is relatively little known about serpins in chickens. In this study, we investigated the expression patterns of serpin genes in the adult chicken oviduct and also the neonatal oviduct of 37-day-old chicks which were treated with an estrogen analogue, diethylstilbestrol (DES). As results SERPINB3 and SERPINB11 genes were highly expressed in DES-treated oviducts but not in non-treated oviduct. Localization of SERPINB3 and SERPINB11 transcripts were specifically found in the gland-like area of adult chicken oviduct. Collectively, our current data suggest that SERPINB3 and SERPINB11 may have distinct functions in the differentiation processes dependent of steroid hormone regulation in the hen oviduct.

S-adenosylhomocysteine Hydrolase-like Protein 1 (AHCYL1) is Mediated by Estrogen-induced ERK1/2 MAPK Cell Signaling and microRNA Regulation to Effect Functional aspects of the Avian Oviduct

Wooyoung Jeong,JinyoungKim,SuzieE.Ahn,WhasunLim,Chulhong Lim,Jinyoung Lee,Seungmin Bae,FullerW.Bazer,GwonhwaSong 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s

S-adenosylhomocysteine hydrolase-like protein 1 (AHCYL1), also known as IP3 receptor- binding protein released with IP3 (IRBIT), regulates IP3-induced Ca2+ release in the cytoplasm of cells and, therefore, is likely to be an important gene regulating various biological processes in the oviduct of chickens. However, the identification of the AHCYL1 gene in chickens has not been investigated. Therefore, the objectives of this study were to examine the tissue- and cell-specific expression of AHCYL1 gene in chicken organs, especially in reproductive organ, and determine functional actions of AHCYL1 in chicken oviduct development via estrogen. The results indicated that AHCYL1 mRNA is expressed in chicken reproductive organs and DES(diethylstilbesterol, a synthetic estrogen agonist) stimulates the cell specific expression of AHCYL1 in immature chicken oviduct. These results suggest that AHCYL1 is a novel estrogen-stimulated gene associated with development of the chicken oviduct. Next, in the present study, we show that inhibition of Erk1/2 can block DES-induced AHCYL1 expression. Also, we found that knockdown of AHCYL1 expression down-regulates expression of oviduct specific genes and AHCYL1 expression is regulated at the post-transcriptional level by specific miRNAs. These results strongly suggest that estrogen-mediated AHCYL1 gene expression plays a crucial role in growth, differentiation and function of the hen oviduct. Also, our results will be useful for understanding the fundamental mechanism(s) of estrogen action responsible for development of hen oviduct. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010-0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

Induction of Apoptosis in Chicken Oviduct Cells by C2-Ceramide

김성학,김상훈,Ji Young Choi,Choon-Ryoul Sihn,서은정,Sun Young Kim,최강덕,Iksoo Jeon,한재용,김태윤 한국분자세포생물학회 2005 Molecules and cells Vol.19 No.2

The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin and its cells undergo cell proliferation and differentiation. There has been no study of the cellular mechanism involved in cell death in the chicken oviduct. Therefore, this study has focused on the study of apoptosis in primary oviduct cells. Because ceramide is known to activate apoptosis in tumor cells and is produced in the oviduct, we used an exogenous ceramide analog to induce cell death. The viability of ceramide-treated chicken oviduct cells decreased in a dose-dependent manner and apoptotic cells were detected by staining with annexin V. The expression of apoptosis-related genes was assessed by RT-PCR and bcl-2 mRNA was found to decrease after exposure to ceramide while Bcl-x mRNA increased 12 h post-treatment. In addition, caspase-3 was expressed strongly in the early stages of apoptosis,while caspase-1 and -9 transcripts increased at later times. We conclude that ceramide induces apoptosis in oviduct-derived primary cells via a caspase- and bcl-2-dependent pathway.

Hormonal regulation of beta-catenin during development of the avian oviduct and its expression in epithelial cell-derived ovarian carcinogenesis

Bae, Seung-Min,Lim, Whasun,Jeong, Wooyoung,Lee, Jin-Young,Kim, Jinyoung,Han, Jae Yong,Bazer, Fuller W.,Song, Gwonhwa Elsevier 2014 Molecular and cellular endocrinology Vol.382 No.1

<P><B>Abstract</B></P> <P>Beta-catenin (CTNNB1) is a dual function molecule that acts as a key component of the cadherin complex and WNT signaling pathway. It has a crucial role in embryogenesis, tumorigenesis, angiogenesis and progression of metastasis. Recently, it has been suggested that the CTNNB1 complex is a major regulator of development of the mouse oviduct and uterus. However, little is known about the <I>CTNNB1</I> gene in chickens. Therefore, in this study, we focused on the <I>CTNNB1</I> gene in the chicken reproductive tract and hormonal control of its expression in the chicken oviduct. <I>CTNNB1</I> was localized specifically to the luminal and glandular epithelium of the four segments of chicken oviduct and DES (diethylstilbestrol, a synthetic non-steroidal estrogen) increased its expression primarily in LE of the magnum. In addition, CTNNB1 mRNA and protein were expressed abundantly in glandular epithelium of endometrioid-type ovarian carcinoma, but not in normal ovaries. Moreover, <I>CTNNB1</I> expression was post-transcriptionally regulated via its 3′-UTR by binding with target miRNAs including <I>miR-217</I>, <I>miR-1467</I>, <I>miR-1623</I> and <I>miR-1697</I>. Collectively, these results indicate that <I>CTNNB1</I> is a novel gene regulated by estrogen in epithelial cells of the chicken oviduct and that it is also abundantly expressed in epithelial cells of endometrioid-type ovarian carcinoma suggesting that it could be used as a marker for diagnosis of ovarian cancer in laying hens and women.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We examine hormonal and post-transcriptional regulation of chicken CTNNB1. </LI> <LI> We evaluate <I>CTNNB1</I> gene expression in cancerous ovaries of laying hens. </LI> <LI> CTNNB1 is a novel estrogen-induced gene during chick oviduct development. </LI> <LI> CTNNB1 is a post-transcriptionally regulated gene by specific microRNAs. </LI> <LI> CTNNB1 is abundant in the endometrioid-type ovarian carcinomas of laying hens. </LI> </UL> </P>

동면기 동안에 한국산 관박쥐 (Rhinolophus ferrumequinum korai) 자성 생식도관 내의 정자저장과 소멸

이정훈,손성원,Lee, Jung-Hun,Son, Sung-Won 한국현미경학회 2000 Applied microscopy Vol.30 No.1

동면기 (11월부터 3월까지) 동안의 한국산 관박쥐(Rhinolophus ferrumequinum korai) 자성 생식도관 내의 정자저장, 정자 생존 여부 및 정자이동을 알아보기 위하여 전자현미경으로 관찰한 결과는 다음과 같았다. (1) 자궁내강, 자궁선내의 정자들은 다수의 백혈구들에 의해 포식되고 소멸되었다. (2)정자들은 수란관 미측 협부에서만 저장되었고, 정자들의 두부는 상피세포쪽을 향하고 있었다. 이는 교미기(10월 초-중순경)에 사출된 정자들이 긴 동면기 동안에 수란관 미측 협부에서만 생존 가능함을 의미하며, 수란관의 미측 협부가 정자의 수정능 획득(capacitation)에 필요한 최적의 장소임을 의미한다. (3) 동면후기인 3월의 수란관 미측협부에는 정자들이 관찰되지 않았다. 이는 배란기인 4월에 난자를 만나기 위해 정자들이 수정부위인 팽대부 쪽으로 이행하였음을 의미한다. 이상의 결과로 미루어 보아, 긴 동면기 동안의 자성 생식도관 내에서의 정자의 장기저장과 생존여부 및 정자이동은 수정을 위한 일종의 mechanism이라 여겨진다. This study was carried out to investigate sperm storage, and the fate of spermatozoa in the female reproductive tract during hibernation in Korean greater horseshoe bat, Rhinolophus ferrumequinum korai. (1) Numerous sperm occurring in uterine lumen and glands were engulfed, and disappeared by the polymorphouclear leucocytes during the hibernation. (2) The stored sperm present in caudal isthmus of oviduct only, the heads of sperm toward the oviductal epithelial cells. Therefore, the projected sperm during the mating season are only alive in the caudal isthmus of oviduct in the long hibernation. The present result suggests that the caudal isthmus of oviduct may play an important role as the principal storage site in capacitation of sperm. (3) In March, the sperm do not occur in the caudal isthmus of oviduct. It suggests that the stored sperm in the caudal isthmus of oviduct should migrate to the ampulla of the site of fertilization to meet ovum in the period of ovulation. The results of this experiment consider that prolonged sperm storage, fate of sperm and sperm migration in the long hibernation have a kind of mechanism for the fertilization.

Aquaporin 3 is regulated by estrogen in the chicken oviduct and is involved in progression of epithelial cell-derived ovarian carcinomas

Yang, C.,Lim, W.,Bae, H.,Song, G. Domendo, Inc ; Elsevier Science Pub. Co 2016 Domestic animal endocrinology Vol.55 No.-

<P>Aquaporins (AQPs) are membrane proteins that passively deliver water across the plasma membrane to play an important role in maintaining cell shape. Members of the AQP family are distributed in most of the tissues in the human body and perform a variety of functions based on the water homeostasis suitable for each organ. However, there is little known about the expression and regulation of AQP family members in chickens. Therefore, we determined the expression of AQPs in various tissues of chickens. Among 13 isotypes, AQP3 was highly expressed in the chicken oviduct. Expression of AQP3 messenger RNA (mRNA) increased in the magnum (P < 0.001) and isthmus (P < 0.001) of chick oviducts treated with diethylstilbestrol. Consistent with these results, the localization of AQP3 was detected in the glandular and luminal epithelia of the magnum and isthmus of oviducts of diethylstilbestrol-treated chicks. In addition, the pattern of expression of AQP3 changed in an estrogen-dependent manner during the molting period. During the regenerative period of the oviduct after molting, expression of AQP3 mRNA increased coordinately with increasing concentrations of estradiol (P < 0.001), whereas expression of AQP3 mRNA decreased as concentrations of estradiol in plasma decreased in response to induced molting (P < 0.001). Also, expression of the AQP3 increased (P < 0.001) in cancerous ovaries of laying hens. In conclusion, AQP3 does not simply function to transport water into and out of cells but also appears to be closely involved in development of the chicken oviduct, which is regulated by estrogens. Furthermore, our results suggest AQP3 as a new diagnostic for early detection and treatment of epithelial cell-derived ovarian carcinomas. (C) 2016 Elsevier Inc. All rights reserved.</P>