• 한국동물학회지
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• 제39권1호
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• pp.115-121
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• 1996

임신중기에서 발기에 이르는 흰쥐의 태반에서 Placental Lactogen I (PL-I), II 그리고 Pit-1 의 유전자 발현 변화를 Northern blot hybridization으로 조사하였다. 그 결과, 임신시기에 따라 PL-I과 PL-II의 mRNA 양과 크기에 변화가 나타났다. 이들 유전자 발현에 미치는 에스트로겐의 영향을 조사하기 위하여, 임신 14일째 쥐의 난소를 제거하고(OVX), 이후 매일 에스트로겐을 투여한 후 (OVX+E), 임신 18일째 태반을 회수하여 PL-I, II 그리고 Pit-1의 유전자 발현을 Northern blot hybridization으로 조사하였다. OVX 군의 경우, PL-I의 mRNA 크기는 1 kb에서 1.3 kb로 PL-II의 mRNA는 0.6kb에서 1 kb로 변화하였다. OVX+E 군에서는 PL-I과 PL-II의 mRNA가 정상대조군과 같은 상태로 환원하였다. 정상대조군에 비하여 OVX와 OVX+E 군에서 PL-I과 PL-II의 mRNA 크기에는 영향을 미치지 못한 반면, mRNA 양은 난소제거시 감소하였다가, 에스트로겐을 투여하면, 부분적으로 회복되는 경향을 보였다. 이러한 본 실험의결과는 에스트로겐이 PL-I과 PL-II의 RNA splicing이나 polyadenylation 등을 포함한 유전자 발현에 영향을 미치며, Pit-1이 이 과정에 개입하는 것을 시사하는 것이다.

• Molecules and Cells
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• 제39권5호
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• pp.403-409
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• 2016

NME1 is a well-known metastasis suppressor which has been reported to be downregulated in some highly aggressive cancer cells. Although most studies have focused on NME1, the NME1 gene also encodes the protein (NME1L) containing N-terminal 25 extra amino acids by alternative splicing. According to previous studies, NME1L has potent anti-metastatic activity, in comparison with NME1, by interacting with $IKK{\beta}$ and regulating its activity. In the present study, we tried to define the role of the N-terminal 25 amino acids of NME1L in $NF-{\kappa}B$ activation signaling. Unfortunately, the sequence itself did not interact with $IKK{\beta}$, suggesting that it may be not enough to constitute the functional structure. Further construction of NME1L fragments and biochemical analysis revealed that N-terminal 84 residues constitute minimal structure for homodimerization, $IKK{\beta}$ interaction and regulation of $NF-{\kappa}B$ signaling. The inhibitory effect of the fragment on cancer cell migration and $NF-{\kappa}B$-stimulated gene expression was equivalent to that of whole NME1L. The data suggest that the N-terminal 84 residues may be a core region for the anti-metastatic activity of NME1L. Based on this result, further structural analysis of the binding between NME1L and $IKK{\beta}$ may help in understanding the anti-metastatic activity of NME1L and provide direction to NME1L and $IKK{\beta}$-related anti-cancer drug design.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.97-97
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• 2003

For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.

• The Korean Journal of Physiology and Pharmacology
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• 제24권3호
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• pp.203-212
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• 2020

Promyelocytic leukemia (PML) gene, through alternative splicing of its C-terminal region, generates several PML isoforms that interact with specific partners and perform distinct functions. The PML protein is a tumor suppressor that plays an important role by interacting with various proteins. Herein, we investigated the effect of the PML isoforms on oncostatin M (OSM)-induced signal transducer and activator of transcription-3 (STAT-3) transcriptional activity. PML influenced OSM-induced STAT-3 activity in a cell type-specific manner, which was dependent on the p53 status of the cells but regardless of PML isoform. Interestingly, overexpression of PML exerted opposite effects on OSM-induced STAT-3 activity in p53 wild-type and mutant cells. Specifically, overexpression of PML in the cell lines bearing wild-type p53 (NIH3T3 and U87-MG cells) decreased OSM-induced STAT-3 transcriptional activity, whereas overexpression of PML increased OSM-induced STAT-3 transcriptional activity in mutant p53-bearing cell lines (HEK293T and U251-MG cells). When wild-type p53 cells were co-transfected with PML-IV and R273H-p53 mutant, OSM-mediated STAT-3 transcriptional activity was significantly enhanced, compared to that of cells which were transfected with PML-IV alone; however, when cells bearing mutant p53 were co-transfected with PML-IV and wild-type p53, OSM-induced STAT-3 transcriptional activity was significantly decreased, compared to that of transfected cells with PML-IV alone. In conclusion, PML acts together with wild-type or mutant p53 and influences OSM-mediated STAT-3 activity in a negative or positive manner, resulting in the aberrant activation of STAT-3 in cancer cells bearing mutant p53 probably might occur through the interaction of mutant p53 with PML.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7315-7319
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• 2013

Background: The metastasis gene osteopontin (OPN) is subject to alternative splicing, which yields three messages, osteopontin-a, osteopontin-b and osteopontin-c. Osteopontin-c is selectively expressed in invasive, but not in noninvasive tumors. In the present study, we examined the expression of OPN-c in esophageal squamous cell carcinomas (ESCCs) and assessed its value as a diagnostic biomarker. Methods: OPN-c expression was assessed by immunohistochemistry in 63 ESCC samples and correlated with clinicopathologic factors. Expression was also examined in peripheral blood mononuclear cells (PBMCs) from 120 ESCC patients and 30 healthy subjects. The role of OPN-c mRNA as a tumor marker was investigated by receiver operating characteristic curve (ROC) analysis. Results: Immunohistochemistry showed that OPN-c was expressed in 30 of 63 cancer lesions (48%)and significantly associated with pathological T stage (P=0.038) and overall stage (P=0.023). Real time PCR showed that OPN-c mRNA was expressed at higher levels in the PBMCs of ESCC patients than in those of healthy subjects (P<0.0001) with a sensitivity as an ESCC biomarker of 86.7%. Conclusion: Our findings suggest that expression of OPN-c is significantly elevated in ESCCs and this upregulation could be a potential diagnostic marker.

• Fisheries and Aquatic Sciences
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• 제14권2호
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• pp.93-104
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• 2011

Two hepcidin paralogs (ojhamp1 and ojhamp2) were isolated and characterized from a euryhaline Javanese ricefish (Oryzias javanicus: Beloniformes). The ojhamp1 cDNA encoded 90 or 91 amino acids (aa) of a typical HAMP1 preproprotein. This preproprotein is believed to cleave and yield the 66 or 67 aa-proprotein, followed by the 26 aa-mature peptide, composed of 8 conserved cysteine residues and the QSHL amino terminal motif. The ojhamp2 cDNA encoded 89 aa of HAMP2 preproprotein, cleaved to yield a 65 aa proprotein, and subsequently the 25 aa-mature peptide. The mature OJHAMP1 possessed a cationic isoelectric point (pI), whereas OJHAMP2 had an anionic charge. At the genomic level, both ojhamp1 and ojhamp2 share a conserved tripartite structure (three exons interrupted by two introns) with other vertebrate hepcidin genes. However, the ojhamp1 was shown to exist as two distinct mRNA species, encoding 90 or 91 aa, due to alternative splicing at the junction site between intron I and exon II. Both ojhamp1 and ojhamp2 transcripts were detected in a wide range of tissue types with varying levels of basal expression, although the highest expression was observed in the liver for both isoforms. Transcriptional response to bacterial challenge using Edwardsiella tarda showed that ojhamp1 was moderately upregulated in the liver but remained unchanged in the kidney. However, the ojhamp2 was significantly suppressed in both the kidney and liver, suggesting a potential diversification between the two paralogs.

• 대한의생명과학회지
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• 제15권4호
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• pp.301-307
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• 2009

Whole-body insulin resistance results largely from impaired insulin-stimulated glucose disposal in skeletal muscle. Our previous studies using differential display and quantitative real-time RT-PCR have shown that a novel cDNA band (DD23) had a higher level of expression in insulin resistant skeletal muscle and it was correlated with whole-body insulin action, independent of age, sex, and percent body fat. In this study, we cloned and characterized DD23. The DD23 sequence is part of the 3'UTR region of the RNA binding motif, single stranded interacting protein (RBMS3). We have cloned the full length cDNA for RBMS3 and identified two splice variants. These variants named DD23-L and DD23-S have 15 and 14 exons respectively and differ from RBMS3 in the 3'UTR significantly. Northern blot analyses showed that an ~8.8 kb mRNA transcript of DD23 was predominantly expressed in skeletal muscle and to a lesser extent in placenta, but not in heart, brain, lung, liver, or kidney, unlike RBMS3. Elevated expression levels of these novel alternatively spliced variants of RBMS3 in skeletal muscle may play a role in whole body insulin resistance.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.114-114
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• 2002

포유동물에서 뇌와 부신에서 합성.분비되는 카테콜아민(Catecholamine, CA)계 신경전달물질인 dopamine(DA), norepinephrine(NE), epinephrine(E)은 체내 각종 생리현상의 조절에 필수적이며, 생식과 관련지어서는 시상하부-뇌하수체 간 GnRH-gonadotropin 호르몬 축의 활성을 조절하는 기능 외에도 번식과 관련된 여러 행동양식을 조절함이 잘 알려져 있다. 본 연구는 CA 생합성 효소들인 tyrosine hydroxylase(TH), dopamine beta-hydroxylase(DBH), phenylethanolamine-N-methyltransferase(PNMT)의 유전자 발현에 미치는 sex steroid의 영향을 조사하였다. 성숙한 암컷 횐쥐(SD strain)의 난소를 제거하고 1주 경과 후 vehicle(sesame oil; OVX+Oil 실험군) 또는 estradiol 17$\beta$(235ug/m1; OVX+E$_2$실험군)이 든 silastic capsule(길이 14mm; 내경 1.55mm; 외경 3.125mm)을 48 시간 동안 처리한 뒤 희생시켰다. 적출된 조직으로부터 RNA를 추출한 후 semi-quantitative RT-PCR을 시행하였다. (i) TH의 발현 정도는 OVX+Oil 군에서는 시상하부) substantia nigra(SNc)) 부신 순으로, OVX+E$_2$군에서는 SN글 부신) 시상하부 순으로 나타났다. TH 발현에 미치는 estradiol의 효과로 SNc과 부신에서는 유의한 증가를 보인데 비해 시상하부에서는 유의한 감소를 관찰하였다. (ii) DBH 발현 정도는 OVX+Oil군에서는 SNc> 부신> 시상 하부 순으로, OVX+E$_2$군에서는 부신＞ SNc＞ 시상하부 순이었다. DBH 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 감소하는 경향을 보였다. (iii) PNMT의 발현의 경우 SNc와 시상하부에서는 기보고된 바와 같이 alternative splicing에 의해 110bp 차이의 크고 작은 두 형태의 cDNA(PNMTI & PNMTs)가 증폭되었으나 부신에서는 작은 cDNA 만이 관찰되었다. PNMTs의 발현 정도는 OVX+Oil군과 OVX+E$_2$군에서 공히 부신＞ 시상하부＞ SNc 순이었고, PNMTI의 발현은 SNc가 시상하부 보다 다소 높은 경향이었으나 유의성은 없었다. PNMTs 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 증가하는 경향을 보였다. 본 연구에서는 CA 생합성 효소들의 유전자 발현의 조절에 미치는 estrogen 의 영향이 세포 기원이 neural crest cell인 부신 수질은 물론 뇌의 상이한 지역간에서도 조직특이적임을 관찰하였다. 이러한 결과는 각 조직에서의 estrogen 수용체 유형의 차이 혹은 작용 모드와 각 효소 유전자 발현 사이에 중요한 상관관계가 있음을 시사한다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권10호
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• pp.1291-1298
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• 2010

The aim of this research was to investigate the dynamic changes of the level of total prolactin receptor (PRLR) mRNA and the short form prolactin receptor (S-PRLR) mRNA in skin of cashmere goats from the initiation of cashmere fibre growth to active growth. Eighteen half-sib wethers were allocated randomly to two groups. Melatonin implants were used in order to initiate growth of cashmere fibre before the normal time and reduce blood plasma prolactin (PRL) concentration. Real-time reverse transcription quantitative polymerase chain reaction (real-time PCR) was used to determine PRLR mRNA expression levels of skin from June to November. The results showed that, in Chinese Inner Mongolia cashmere goats, there were seasonal variations in expression of total PRLR mRNA in skin with levels decreasing from June to October. Synchronously, the cashmere fibre growth rate gradually increased during this period, but the expression levels of S-PRLR mRNA did not decrease along with seasonal variation from initiation to active growth of cashmere fibre. These results suggest that expression levels of S- PRLR mRNA might be involved in the process of cashmere growth. It was also possible that the change of alternative splicing of PRLR occurred in the skin of cashmere goats from proanagen to anagen.

• Asian-Australasian Journal of Animal Sciences
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• 제18권7호
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• pp.1017-1021
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• 2005

Myostatin is a negative regulator of skeletal muscle growth and a loss of functional myostatin protein increases muscle hypertrophy and hyperplasia in cattle. The present study was conducted to investigate whether maternal passive immunization against myostatin would improve growth performance in chickens. A complete broiler myostatin cDNA was cloned and it was expressed into two transcripts as 1,128 bp and 985 bp by alternative splicing. A conjugated mature myostatin (350 bp) was used to induce autoimmunization and maternal passively immunized chickens was used for the experiment. It was confirmed that there was a maternal passive immunization against myostatin at zero weeks of age, but its effect was reduced by 6 weeks of age. The auto-immunized groups showed smaller body weights than those of control group during the growing period and the difference was getting bigger with time until 6 weeks of age. These results suggest that passive autoimmunization against myostatin used in this study is not potent enough to stimulate growth performance in chickens.