Identification and characterization of ERRFI1 in chickens

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

ERBB receptor feedback inhibitor 1 (ERRFI1, also known as MIG-6) is a negative regulator of epidermal growth factor signaling pathway in various biological processes. However, the identification and characterization of ERRFI1 gene in chickens has not been reported. In the present study, we investigated ERRFI1 gene expression in each embryonic stage and adult organs by RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemical analyses. Our current data showed that the ERRFI1 mRNA was expressed in brain, kidney, small intestine and muscle of male embryo, and weakly expressed in kidney, heart, liver and muscle of female embryo. In adult tissues, ERRFI1 is specifically expressed in seminiferous tubules in the testis and glandular epithelium in the oviduct after sexual maturation. Immunohistochemistry revealed that ERRFI1 protein was particularly abundant in the nuclei of luminal and glandular epithelia in the oviduct. In addition, treatment with synthetic estrogen analog diethylstilbestrol (DES) stimulated ERRFI1 expression in the oviduct. Collectively, the present studies validate ERRFI1 as a novel estrogen-stimulated gene in the oviductal epithelia in chickens.

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.

Estrogen Regulates ERRFI1 Expression during Development of the Chicken Oviduct

Chul-Hong Lim(임철홍),Suzie E. Ahn,Whasun Lim,Wooyoung Jeong,Jinyoung Kim,Fuller W.Bazer,Jae Yong Han,Gwonhwa Song 한국가금학회 2011 한국가금학회 정기총회 및 학술발표회 Vol.28 No.-

Matrix metalloproteinase 3 is a stromal marker for chicken ovarian cancer

CHOI, JIN WON,AHN, SUZIE E.,RENGARAJ, DEIVENDRAN,SEO, HEE WON,LIM, WHASUN,SONG, GWONHWA,HAN, JAE YONG Spandidos Publications 2011 Oncology letters Vol.2 No.6

<P>Matrix metalloproteinases (MMPs) are involved in the degradation of the extracellular matrix and basement membranes. Due to this, MMPs have been thought to promote invasion and metastasis of cancer cells and angiogenesis in tumors. Even though the chicken is a useful animal model for studying human ovarian cancer, no reports exist of the MMP expression pattern in chicken ovarian cancer. Therefore, we investigated the expression pattern of MMPs in chicken ovarian cancer. Results of RT-PCR and quantitative RT-PCR analyses showed MMP3 to be over-expressed in cancerous hen ovaries. In situ hybridization analysis of cancerous chicken ovaries showed that MMP3 mRNA was predominantly localized in the stroma, which is similar to MMP3 expression in human cancers. The results suggest that the expression pattern of MMP3 mRNA in chicken ovarian cancer is similar to that in various types of human cancer. Moreover, MMP3 potentially plays a significant role in developing ovarian cancer in chickens. The cell type-specific expression of MMP3 makes this gene a unique marker for ovarian cancer in chickens.</P>

Chicken SERPINBs: Characteristics, Tissue Specific Expression and Regulation of Expression by Estrogen

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

Serpins are a superfamily of related protease inhibitors with common structural features and inhibitory mechanisms. However, SERPINA 14 in mammals does not have inhibitory activity against most known proteases. Rather, it may have an immunoregulatory role in mammals to prevent rejection of the fetal allograft by inhibiting lymphocyte proliferation and natural killer cell function. In the pig, SERPINA14 is involved in iron transport to the fetus by binding to and stabilizing the iron-binding protein uteroferrin (ACP5). In chickens, these very little known about serpins in chickens. Therefore, we investigated the expression patterns of serpin genes in the oviduct of adult hens and in the oviduct of 37-day-old chicks treated with an estrogen analogue, diethylstilbestrol (DES). Results indicated that SERPINB3 and SERPINB11 genes were highly expressed in oviducts of DES-treated chicks, but not in oviducts of control chicks. Both SERPINB3 and SERPINB11 transcripts were localized specifically to the gland-like areas of oviducts of DES-treated chicks. Immunohistochemical analyses confirmed that SERPINB3 and SERPINB11 proteins were present in the gland-like area and luminal epithelium of the oviducts of DES-treated chicks. Collectively, the results suggest that SERPINB3 and SERPINB11 are expressed in response to estrogens and they have distinct functions related to development and differentiation of the mature oviduct in hens.

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.

Avian SERPINB11 gene: characteristics, tissue-specific expression, and regulation of expression by estrogen.

Lim, Whasun,Kim, Ji-Hye,Ahn, Suzie E,Jeong, Wooyoung,Kim, Jinyoung,Bazer, Fuller W,Han, Jae Yong,Song, Gwonhwa Society for the Study of Reproduction [etc.] 2011 BIOLOGY OF REPRODUCTION Vol.85 No.6

<P>Serpins, a group of proteins with similar structural and functional properties, were first identified based on their unique mechanism of action: their inhibition of proteases. While most serpins have inhibitory roles, certain serpins are not involved in canonical proteolytic cascades but perform diverse functions including storage of ovalbumin in egg white, transport of hormones (thyroxine- and cortisol-binding globulin), and suppression of tumors. Of these, serpin peptidase inhibitor, clade B, member 11 (SERPINB11) is not an inhibitor of known proteases in humans and mice, and its function is unknown. In the present study, the SERPINB11 gene was cloned, and its expression profile was analyzed in various tissues from chickens. The chicken SERPINB11 gene has an open reading frame of 1346 nucleotides that encode a protein of 388 amino acids that has moderate homology (38.8%-42.3%) to mammalian SERPINB11 proteins. Importantly, SERPINB11 mRNA is most abundant in the chicken oviduct, specifically luminal and glandular epithelia, but it was not detected in any other chicken tissues of either sex. We then determined effects of diethylstilbestrol (DES; a synthetic nonsteroidal estrogen) on SERPINB11 expression in the chicken oviduct. Treatment of young chicks with DES induced SERPINB11 mRNA and protein only in luminal and glandular epithelial cells of the oviduct. Collectively, these results indicate that the novel estrogen-induced SERPINB11 gene is expressed only in epithelial cells of the chicken oviduct and implicate SERPINB11 in regulation of oviduct development and differentiated functions.</P>

Avian SERPINB11 gene: a marker for ovarian endometrioid cancer in chickens.

Lim, Whasun,Kim, Ji-Hye,Ahn, Suzie E,Jeong, Wooyoung,Kim, Jinyoung,Bazer, Fuller W,Han, Jae Yong,Song, Gwonhwa The Society 2012 Experimental biology and medicine Vol.237 No.2

<P>As serine and cysteine proteinase inhibitors, serpins, such as SERPINB5, cause ovarian, colorectal and pancreatic adenocarcinomas. We identified SERPINB11 as a novel estrogen-induced gene in chickens during oviduct development. The chicken is a unique animal model for research on human ovarian cancer, because it spontaneously develops epithelial cell-derived ovarian cancer as in women. Therefore, this study investigated the expression pattern, CpG methylation status, and miRNA regulation of the SERPINB11 gene in normal and cancerous ovaries from chickens. Our results indicate that SERPINB11 is most abundant in the glandular epithelium of endometrioid adenocarcinoma of cancerous, but not normal, ovaries of hens. In addition, bisulfite sequencing revealed that about 30% of -110 CpG sites are methylated in ovarian cancer cells, whereas -110 CpG sites are demethylated in normal ovarian cells. Next, we determined whether miR-1582 influences SERPINB11 expression via its 3'UTR and found that it does not directly target the 3'UTR of SERPINB11 mRNA. Therefore, it is unlikely that post-transcriptional regulation influences SERPINB11 expression in the chicken ovary. On the other hand, in human ovarian cancer cells such as OVCAR-3, SKOV-3 and PA-1 cells, immunoreactive SERPINB11 protein was predominant in the cytoplasm and had a similar expression pattern to that in chicken ovarian cancer cells. Collectively, these results suggest that SERPINB11 is a biomarker for chicken ovarian endometrioid carcinoma that could be used for diagnosis and monitoring effects of therapies for the disease in women.</P>

Cell-specific and temporal aspects of gene expression in the chicken oviduct at different stages of the laying cycle.

Jeong, Wooyoung,Lim, Whasun,Kim, Jinyoung,Ahn, Suzie E,Lee, Hyung Chul,Jeong, Jae-Wook,Han, Jae Yong,Song, Gwonhwa,Bazer, Fuller W Society for the Study of Reproduction [etc.] 2012 BIOLOGY OF REPRODUCTION Vol.86 No.6

<P>Egg formation and embryonic development occur as the yolk passes through the magnum, isthmus, and shell gland of the oviduct before oviposition in hens. The present study identified candidate genes associated with secretory function of the chicken oviduct after ovulation and contributing to egg formation and oviposition. Hens (n = 5 per time point) were euthanized to recover the reproductive tract when the egg was in the magnum (3 h after ovulation) and the shell gland (20 h after ovulation). Total RNA was extracted from each segment of the oviducts and subjected to Affymetrix chicken GeneChip analysis. Quantitative PCR and in situ hybridization analyses of selected genes confirmed the validity of the gene expression patterns detected using microarray analysis. In particular, ACP1, CALB1, CYP26A1, PENK, RCAN1 and SPP1 expression increased significantly in the shell gland between 3 h and 20 h postovulation, whereas only RCNA1 expression increased significantly in the magnum between 3 h and 20 h postovulation. Results of the high-throughput analysis revealed cell-specific and temporal changes in gene expression in the oviduct at 3 h and 20 h postovulation in laying hens provide novel insight into changes at the molecular and cellular levels of candidate genes related to formation of the egg and oviposition.</P>

Increased expression of Gallicin 11 in chicken ovarian cancer

Wooyoung Jeong(정우영),Whasun Lim,Jihye Kim,Suzie E. Ahn,Jae Yong Han,Gwonhwa Song 한국가금학회 2010 한국가금학회 정기총회 및 학술발표회 Vol.27 No.-

Gallicin 11, also known as beta-defensin 11, is a member of the chicken beta-depensin family and constitutes a vitellin membrane outer layer of egg yolk. The protein is an essential component of innate immunity and acts in the antimicrobial process by disrupting the membrane integrity of microbes. However, it has yet to be investigated that expression and functional role of Gallicin 11 in chickens with ovarian cancer. Therefore, the purpose of this study was to investigate the changes of Gallicin 11 expression in chicken with ovarian cancer in order to understand the mechanism(s) responsible for the etiology and development of ovarian cancer in chickens. In the current study, we performed RT-PCR, quantitative RT-PCR, and in situ hybridization analyses to determine the expression, quantitation and localization of Gallicin 11 mRNA in normal and cancerous chicken ovaries. The results showed that Gallicin 11 mRNA level is highly increased in the cancerous ovarian tissue compared to the normal ovary. Interestingly, Gallicin 11 mRNA is particularly abundant in gland area of ovarian cancer tissue but not in other cell-types such as stroma and connective tissues in the oviduct. Our results suggest that Gallicin 11 has important functions in the development of epithelial ovarian cancer and provide the basis for understanding the mechanism(s) responsible for the development of ovarian cancer in chickens.