Expression of Luteinizing Hormone (LH) and Its Receptor Gene in Rat Mammary Gland

류종순,김재만,이성호 한국발생생물학회 2000 발생과 생식 Vol.4 No.2

Gonadotropin-releasing hormone (GnRH)과 그 수용체가 흰쥐의 난소, 정소, 자궁, 태 반 그리고 유선 등의 생식기관에서 발현됨이 알려져 있다. 더욱이, 뇌하수체 전엽에 작용하는 GnRH의 표적 산물로 알려진 luteinizing hormone (LH)이 흰쥐 생식소에서도 발현됨이 알려졌는데, 이는 생식소 내에 GnRH-LH로 이루어진 국부 회로 (local circuit)가 존재함을 시사하는 것이다 본 연구는 LH와 그 수용 Recent studies have clearly shown that the expression of genes for gonadotropin-releasing hormone (GnRH) and its receptor in the rat reproductive organs including ovary, testis, placenta uterus and mammary gland. Moreover, luteinizing hormone (LH) classically known to be a main target product of GnRH in anterior pituitary has been found in rat gonads. These findings suggested the presence of local circuit composed of GnRH and LH in the rat gonads. The present study was undertaken to elucidate whether the genes for LH and its receptor are expressed in rat mammary gland. Expression of LH and its receptor genes in the rat mammary gland was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and specific LH radioimmunoassay (RIA). The LH transcripts in the mammary gland from cycling rats contained the pituitary type of LH exons 1~3 encoding the entire LH polypeptide but lacked the rat testis-specific LH exon(s). Presence of -subunit transcripts in the rat mammary gland were determined by RT-PCR. The cDNA fragments encoding exons 2~7 of rat LH receptor transcripts were amplified in both rat ovary and mammary gland samples. We could detect the GnRH expression in mammary gland from cycling virgin rats, and this result disagreed with previous report that mammary GnRH expression is occured in lactating rats only. Considerable amounts of immunoreactive LH molecules with good RIA parallelism in standard curve were detected in crude extracts from the rat mammary gland, indicating that the immunoreactive LH materials in the gland might be identical to authentic pituitary LH. To our knowledge, the present study demonstrated for the first time the expression of LH subunits and LH receptor in the rat mammary gland. Our findings suggested that the mammary gland might be the novel source and target of LH and the mammary LH could be act as a local regulator with auto-and/or paracrine manner under the regulation of local GnRH.

Identifying long non-coding RNAs and characterizing their functional roles in swine mammary gland from colostrogenesis to lactogenesis

Shi Lijun,Zhang Longchao,Wang Ligang,Liu Xin,Gao Hongmei,Hou Xinhua,Zhao Fuping,Yan Hua,Cai Wentao,Wang Lixian 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.6

Objective: This study was conducted to identify the functional long non-coding RNAs (lncRNAs) for swine lactation by RNA-seq data of mammary gland. Methods: According to the RNA-seq data of swine mammary gland, we screened lncRNAs, performed differential expression analysis, and confirmed the functional lncRNAs for swine lactation by validation of genome wide association study (GWAS) signals, functional annotation and weighted gene co-expression network analysis (WGCNA). Results: We totally identified 286 differentially expressed (DE) lncRNAs in mammary gland at different stages from 14 days prior to (-) parturition to day 1 after (+) parturition, and the expressions of most of lncRNAs were strongly changed from day –2 to day +1. Further, the GWAS signals of sow milk ability trait were significantly enriched in DE lncRNAs. Functional annotation revealed that these DE lncRNAs were mainly involved in mammary gland and lactation developing, milk composition metabolism and colostrum function. By performing weighted WGCNA, we identified 7 out of 12 lncRNA-mRNA modules that were highly associated with the mammary gland at day –14, day –2, and day +1, in which, 35 lncRNAs and 319 mRNAs were involved. Conclusion: This study suggested that 18 lncRNAs and their 20 target genes were promising candidates for swine parturition and colostrum occurrence processes. Our research provided new insights into lncRNA profiles and their regulating mechanisms from colostrogenesis to lactogenesis in swine. Objective: This study was conducted to identify the functional long non-coding RNAs (lncRNAs) for swine lactation by RNA-seq data of mammary gland.Methods: According to the RNA-seq data of swine mammary gland, we screened lncRNAs, performed differential expression analysis, and confirmed the functional lncRNAs for swine lactation by validation of genome wide association study (GWAS) signals, functional annotation and weighted gene co-expression network analysis (WGCNA).Results: We totally identified 286 differentially expressed (DE) lncRNAs in mammary gland at different stages from 14 days prior to (-) parturition to day 1 after (+) parturition, and the expressions of most of lncRNAs were strongly changed from day –2 to day +1. Further, the GWAS signals of sow milk ability trait were significantly enriched in DE lncRNAs. Functional annotation revealed that these DE lncRNAs were mainly involved in mammary gland and lactation developing, milk composition metabolism and colostrum function. By performing weighted WGCNA, we identified 7 out of 12 lncRNA-mRNA modules that were highly associated with the mammary gland at day –14, day –2, and day +1, in which, 35 lncRNAs and 319 mRNAs were involved.Conclusion: This study suggested that 18 lncRNAs and their 20 target genes were promising candidates for swine parturition and colostrum occurrence processes. Our research provided new insights into lncRNA profiles and their regulating mechanisms from colostrogenesis to lactogenesis in swine.

Retinoic acid signaling and the initiation of mammary gland development

Cho, K.W.,Kwon, H.J.,Shin, J.O.,Lee, J.M.,Cho, S.W.,Tickle, C.,Jung, H.S. Academic Press 2012 Developmental Biology Vol.365 No.1

Retinoic acid receptors (RARs), which are involved in retinoic acid signal transduction, are essential for maintaining the differentiated state of epithelial tissues. Mammary glands are skin appendages whose development is initiated through continuous cell-cell interactions between the ectoderm and the adjacent mesenchyme. Considerable progress has been made in elucidating the molecular basis of these interactions in mammary gland formation in mouse embryos, including the network of initiating signals comprising Fgfs, Wnts and Bmps involved in gland positioning and the transcription factors, Tbx3 and Lef1, essential for mammary gland development. Here, we provide evidence that retinoic acid signaling may also be involved in mammary gland development. We documented the expression of gene-encoding enzymes that produce retinoic acid (Raldh2) and enzymes that degrade it (Cyp26a1, Cyp26b1). We also analyzed the expression of RAR-β, a direct transcriptional target of retinoic acid signaling. Raldh2 and RAR-β were expressed in E10-E10.5 mouse embryos in somites adjacent to the flank region where mammary buds 2, 3 and 4 develop. These expression patterns overlapped with that of Fgf10, which is known to be required for mammary gland formation. RAR-β was also expressed in the mammary mesenchyme in E12 mouse embryos; RAR-β protein was expressed in the mammary epithelium and developing fat pad. Retinoic acid levels in organ cultures of E10.5 mouse embryo flanks were manipulated by adding either retinoic acid or citral, a retinoic acid synthesis inhibitor. Reduced retinoic acid synthesis altered the expression of genes involved in retinoic acid homeostasis and also demonstrated that retinoic acid signaling is required for Tbx3 expression, whereas high levels of retinoic acid signaling inhibited Bmp4 expression and repressed Wnt signaling. The results of the experiments using RNAi against Tbx3 and Wnt10b suggested feedback interactions that regulate retinoic acid homeostasis in mammary gland-forming regions. We produced a molecular model for mammary gland initiation that incorporated retinoic acid signaling.

개에서 땀샘 선암종과 유선 복합형 선종의 동시 발생

박지성,정지열,조숙희,정종태,강태영,김재훈,Park, Ji-Sung,Jung, Ji-Youl,Jo, Suk Hee,Cheong, Jongtae,Kang, Tae-Young,Kim, Jae-Hoon 대한수의학회 2008 大韓獸醫學會誌 Vol.48 No.3

A 7-year-old female Shih Tzu dog with lots of masses in the whole mammary gland was presented to the surgery department of the Veterinary Teaching Hospital in the Cheju National University. After surgical excision, all mammary samples were referred to Pathology Department of Veterinary Medicine. Grossly, masses were measuring up to $6.5{\times}4{\times}1cm$ and on cut surface of masses in right 1st, 3rd, 4th, 5th and left 1st, 3rd, 4th, 5th mammary masses were well delineated and firm, sulphur yellow, solid round to oval shape. Microscopically, most neoplastic sweat glands were severely proliferated in dermis and subcutis. Most tubules were lined by round to oval shaped epithelium with eosinophilic cytoplasm, hyperchromatic nuclei with high mitotic figures and severe central necrosis. The neoplastic epithelium also had PAS-positive diastase-resistant cytoplasmic granules, but negative with Perls iron stain. The left 2nd mass was well delineated, and had several dark brown areas and yellowish white glittered areas. Mass was well circumscribed with dense connective tissue. Neoplastic areas contained irregular sized mammary gland with papillary grown luminal epithelial cells in single or double cells layer with mitotic figures and small amounts of proliferated myoepithelial cells. Proliferated myoepithelial cells also produced slightly basophilic mucinous materials. Based on the gross, histopathologic and special staining characteristics, this dog was diagnosed as 90% of apocrine sweat gland aenocarcinoma and 10% mammary. complex adenomas in mammary masses. In our best knowledge, this is the first report for concurrent occurrence of apocrine sweat gland adenocarcinoma and mammary gland complex adenoma in mammary masses of the same dog.

Physiological Regulation of Luteinizing Hormone(LH) Expression in Rat Mammary Gland during Differentiation

이성호 한국발생생물학회 2001 발생과 생식 Vol.5 No.2

태반이나 생식소 등 시상하부 이외의 조직에서도 gonadotropin releasing hormone(GnRH)과 그 수용체가 발현되어 조직 특이적인 기능을 담당함은 잘 알려진 사실이다. 최근 GnRH와 그 수용체 유전자가 흰쥐 유선에서도 발현됨이 증명되었고, LH -와 -subunit와 LH 수용체에 대한 전사체 역시 흰쥐 유선에 존재함이 확인되었다 본 연구는 흰쥐 유선 LH의 발현과 유선의 분화과정 간의 상관관계를 조사하기 위해서 생식주기, 임신 The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

개에서 땀샘 암종, 유선종양 및 방광 이행상피암종의 동시 발생

정연철,우계형,윤영민,김재훈 한국임상수의학회 2016 한국임상수의학회지 Vol.33 No.1

A 7-year-old female Pointer dog with multiple masses in the axilla, mammary gland, and bladder was submitted to the Pathology Department of the College of Veterinary Medicine in the Jeju National University. Grossly, mass between right axilla and 1st mammary gland, 15 × 10 cm in size, was well delineated and firm, slightly soft center, oval shape. And masses in right 1st, 3rd and 5th mammary gland were well delineated and sulphur yellow in color on the cut-surface. Numerous round to oval shaped masses, 0.3 to 2 cm in diameter were existed in the lung. Urinary bladder mucosa had rough and thick and round to oval papillary masses, 0.1 to 2 cm in diameter, on surface. Microscopically, masses in right axilla, 1st mammary gland, lung and axillary lymph node were composed of poorly differentiated tubules originated from apocrine gland. Lining neoplastic epithelium showed high mitotic figures, typical apical secretory blebs, and PAS-positive diastase-resistant cytoplasmic granules. Masses in 3rd and 5th mammary gland were confirmed as mammary complex adenoma and simple adenoma respectively. The masses in the urinary bladder were covered with stratified transitional epithelium with marked cellular atypia and high mitotic figures. Some neoplastic cells showed focal invasion into substantia propria of bladder. Immunohistochemaically, neoplastic transitional epithelium demonstrated positive reactions for cytokeratin 7, AE1/AE3, and MNF116. Based on the gross, histopathologic and immunohistochemical characteristics, this dog was diagnosed as apocrine carcinoma, mammary gland tumor including simple adenoma and complex adenoma and bladder transitional cell carcinoma. And distant metastases of apocrine carcinoma in right axilla were observed in axillary lymph node and lungs. This is the first report for concurrent occurrence of apocrine carcinoma, mammary gland tumor, and transitional cell carcinoma in a same dog.

Expression of ErbB receptors in the pre-pubertal and pubertal virgin mammary glands of dairy cows

Lee, Byung-Woo,Kim, Yo-Han,Jeon, Byung-Suk,Singh, Naresh Kumar,Kim, Won-Ho,Kim, Meing-Jooung,Yoon, Byung-Il The Korean Society of Veterinary Science 2012 大韓獸醫學會誌 Vol.52 No.4

In the present study, we investigated the expression patterns of ErbB family proteins in the pre-pubertal and pubertal mammary glands of dairy cows in association with gland development. For this study, we performed immunohistochemistry for ErbB-1-4 and Ki-67 cell proliferation marker. We found that the pre-pubertal and pubertal mammary glands had typical structures, including ducts and terminal end buds embedded in the stroma, and no development of lobuloalveolar structures. On immunohistochemistry, ErbB-2 and ErbB-3 were strongly expressed in the cytoplasm and nuclei in the epithelial cells of mammary ducts and terminal end buds, and stromal cells, whereas ErbB-1 and ErbB-4 were weakly expressed only in the cytoplasm of gland epithelium and stromal cells, irrespective of the developmental stage. Cell proliferation was inactive in the mammary gland cell compartments in both phases. Thus, expression of the ErbB family in the developing mammary glands was not associated with their functional effects, such as cell proliferation and lobuloalveolar development. In conclusion, ErbB receptors were differentially expressed in the epithelial and stromal cells of virgin mammary glands of dairy cows. Compared with rodent mammary glands, ErbB-3 and ErbB-4 were found to be highly expressed in bovine mammary glands.

Mammary gland tumors in three male dogs

Han, Jeong-Hee,Kim, Kyeong-Soo,Kim, Jae-Hoon The Korean Society of Veterinary Science 2016 大韓獸醫學會誌 Vol.56 No.4

Mammary gland tumors are very rare in male dogs. In this study, four mammary gland tumors from 3 male dogs (2 intact, 1 neutered) were collected from local animal hospitals. The dogs included two purebred Shih Tzu (1 intact, 1 neutered) and one intact purebred Cocker Spaniel. The mean age of dogs with mammary gland tumors was 9 years (5-12 years). Two dogs had a solitary mass, whereas one dog had two mammary masses. Of the four tumor masses, three were observed in the fourth or fifth mammary glands, and one was observed in the third mammary gland. According to histopathologic examinations, all four mammary masses from three dogs were benign tumors including two benign mixed tumors in one case and two complex adenomas. There were no history of obesity, testicular tumors, diabetes, and sex hormonal therapy in any male dogs with mammary tumors. Surgical excision was the only reported treatment for these tumors. No recurrence or metastasis was recorded up to 25 months after surgery.

개 유선종양의 병리학적 관찰 및 유선 혼합종양의 연골기원에 관한 면역조직화학적 연구

김용백,서일복,김재훈,박은정,김대용,한정희,Kim, Yong-baek,Seo, Il-bok,Kim, Jae-hoon,Bak, Eun-jung,Kim, Dae-yong,Han, Jeong-hee 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.4

Sixteen mammary gland tumors were collected from Seoul National University and Kangwon National University. The average age of the bitches with mammary gland tumor was 10 years. Total 17(60.7%) out of 28 tumor masses observed in 4th and 5th glands. Classification of these tumors according to Hampe and Misdorp were simple adenoma, complex adenoma, benign mixed tumor, papillary adenocarcinoma, solid adenocarcinoma and malignant mixed tumor. Immunohistochemical reaction of the intermediate filaments against normal canine mammary gland showed as followed; anti-cytokeratin 18 was strong and anti-cytokeratin 14 was moderate to the luminal epithelium. Anti-cytokeratin 14 and anti-pancytokeratin to the myoepithelium were showed strong, but anti-vimentin was weak in reactivity. Anti-vimentin to the interstitial cells was represented strong reactivity. The origin of cartilage in mixed tumor of canine mammary gland was studied immunohistochemically with antibodies against intermediate filament. In mammary gland mixed tumors, cartilage tumor tissues were surrounded with the irregularly demarcated three zones composed of adjacent star shaped cells in myxoid areas, proliferative spindle shaped cells and basal located proliferated cells. From basal proliferated cells to star shaped cells, the immunohistochemical reactivity of myoepithelium specific anti-pancytokeratin was decreased gradually and the reactivity of interstitial cell specific anti-vimentin was increased gradually. Based on these immunohistochemical staining patterns, we suggested that the origin of cartilagenous components in canine mammary gland mixed tumor is most likely to the proliferation and metaplsia of myoepithelium.

Expression of B Cell Activating Factor Pathway Genes in Mouse Mammary Gland

Choi, S.,Jung, D.J.,Bong, J.J.,Baik, M. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.2

In our previous study, overexpression of extracellular proteinase inhibitor (Expi) gene accelerated apoptosis of mammary epithelial cells, and induced expression of B cell activating factor (BAFF) gene. In this study, we found induction of BAFF-receptor (BAFF-R) gene expression in the Expi-transfected cells. A proliferation-inducing ligand (APRIL) gene is another TNF family member and the closest known relative of BAFF. We found induction of APRIL gene expression in the Expi-overexpressed apoptotic cells. NF-${\kappa}$B gene was also induced in the Expi-overexpressed cells. Expression patterns of BAFF and APRIL pathway-related genes were examined in in vivo mouse mammary gland at various reproductive stages. Expression levels of BAFF gene were very low at early pregnancy, increased from mid-pregnancy, and peaked at lactation, and thereafter decreased at involution stages of mammary gland. Expression of BAFF-R gene was highly induced in involution stages compared to lactation stages. Thus, expression patterns of BAFF-R gene were correlated to apoptotic status of mammary gland: active apoptosis of mammary epithelial cells occurs at involution stage of mammary gland. Expression levels of NF-${\kappa}$B gene were higher in involution stages compared to lactation stages. We analyzed mRNA levels of bcl-2 family genes from different stages of mammary development. Bcl-2 gene expression was relatively constant during lactation and involution stages. There was a slight increase in bcl-xL gene expression in involution stages compared to lactation state. Bax gene expression was highly induced in involution stage. Our results suggest that signaling pathways activated by both BAFF and ARRIL in mammary gland point towards NF-${\kappa}$B activation which causes upregulation of bax.