• Animal cells and systems
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• 제2권2호
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• pp.249-258
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• 1998

Obesity, one of the most common metabolic diseases in industrial countries is characterized by an increase in the number or size of adipocytes. In an effort to create transgenic mouse models for the study of obesity we developed a novel technique in which adipose tissue can be ablated genetically at will, at any specific developmental stage and/or physiological condition, by the treatment of ganciclovir. We made a series of adipocytespecific expression vectors using minimal regulatory regions of brown adipocyte-specific uncoupling protein (UCP-1) gene and adipocyte-specific aP2 gene, and then analyzed their expression characteristics in cultured cell lines. When both constructs pUCP-LacZ and paP2-LacZ were transfected transiently into differentiating 3T3-L1 (pre-while adipocytes) and HIB-1B (pre-brown adipocytes) cell lines in vitro and then monitored by X-gal staining of cells, these regulatory regions were sufficient to show proper differentiation stage-specific expression in adipocvtes. To confirm that adipocytes expressing HSV-TK controlled by these minimal requlatory elements are sufficient to kill themselves with ganciclovir treatment pUCP-TK and paP2-TK expression constructs were transfected stably into HIB-1B and 3T3-L1 cells, respectively, and their ganciclovir sensitivities were tested during in vitro differentiation of cells. As expected more than 80% of cells were dead by the 7th day of treatment with ganciclovir while negative control cells were not affected at all. The data suqqest that the constructed vectors are suitable for obtaining novel obese transqenic models based on a conditional genetic tissue ablation method.

• 생약학회지
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• 제49권3호
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• pp.262-269
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• 2018

Sparassis crispa (Wulf.) is an edible/medicinal mushroom and has been reported to biological activities such as antitumor, anti-angiogenesis, antioxidant and wound healing. However, there have not been many researches on osteoarthritis of S. crispa. The aim of this study was to investigate the effects of S. crispa extract on rats with osteoarthritis induced by MIA. Osteoarthritis is a gradually developmental disease that early stage, causes joint stiffness and complains of joint pain. In addition, it gives rise to edema and hypo-function. The results of this study, S. crispa extract effectively inhibited ROS production, increased the production of antioxidant protein SOD and catalase in knee joint cartilage tissue. In addition, S. crispa extract inhibited the expression of pro-inflammatory cytokines and enzymes such as NOX4 and $P47^{phox}$, which are involved in the expression of COX-2, iNOS and the production of ROS. Also, S. crispa extract inhibited the destruction of synovial tissue, cartilage tissue and proteoglycans in articular cartilage in rats.

• 해부∙생물인류학
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• 제26권1호
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• pp.41-49
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• 2013

치아발생 및 형태형성 과정에서 치아상피와 치아간엽을 구성하는 세포는 동적인 세포주기의 변화가 일어난다. 현재까지 세포증식은 치아발생에 중요한 현상으로 알려져 있지만, 치아발생 중에 일어나는 복잡한 분자적 기전과 연관해서 세포주기의 각 시기가 어떻게 관여하는지에 대해서는 충분한 연구가 이루어지지 않았다. 그러므로 본 연구는 치아발생 기전과 세포주기의 시기의 변화와의 관계를 밝히고자 하였다. 치아발생 과정에서 일어나는 형태변화를 확인하기 위해 싹시기, 모자시기, 종시기의 쥐 앞니 및 어금니 치배를 헤마톡실린-에오신으로 염색하여 조직학적으로 관찰하였다. 또한 세포주기 시기의 표지자인 Ki-67, 싸이클린 A, 싸이클린 D1의 발현양상을 관찰하기 위해 면역조직화학염색을 시행하였다. 싹시기, 모자시기, 종시기에서 증식하는 세포들은 Ki-67과 싸이클린 A를 발현하는 것을 확인하였다. 싸이클린 D1은 앞니의 상아질모세포 및 모자시기의 사기질결절에서 특이적인 발현을 보였으며, 이곳에서는 Ki-67이나 싸이클린 A가 발현되지 않는다는 것을 발견하였다. 본 연구는 치아발생 중 각 주요 시기에서 세포주기의 변화를 관찰하였으며, 이는 치아발생에 관여하는 기전에 대한 중요한 정보를 제공한다. 또한 본 연구의 결과는 지금까지의 앞니의 사기질모세포 및 사기질결절의 특성에 대한 지식을 이해하는 데에 중요한 자료가 될 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제53권3호
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• pp.195-202
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• 2011

Maintenance of adequate telomere length in developing cells is the most important concern to preserve the integrity of the genome. The length of telomere is strictly regulated by numerous telomere-binding proteins and/or interacting factors. Even though the expression of telomerase in the male reproductive tract has been characterized, developmental expressional profiling of telomerase and other telomere-associated proteins has not been determined in detail. The present study was attempted to examine expression patterns of catalytic subunit (Tert) and RNA component (Terc) of telomerase and two telomerase associated factors, telomerase associated protein 1 (Tep1) and TERF1 (TRF1) interacting nuclear factor 2 (Tinf2) in the testis and seminal vesicle of male rat during postnatal development. The real-time PCR analysis was utilized to quantify mRNA expression of molecules. The abundance of Tep1 mRNA in the testis and seminal vesicle was the highest at 5 months of age. Expressional fluctuation of Tinf2 during postnatal development was found in the testis, while expression of Tinf2 in the seminal vesicle was gradually increased until 5 months of age and then significantly decreased later. mRNA level of Tert gene in the testis was significantly increased at the adult and the elder, while the highest expression of Tert gene in the seminal vesicle was found at 5 months of age. Expression of Terc transcript in the testis and seminal vesicle was the highest at 5 months of age, followed by significant reduction at 1 and 2 years of ages. Such differential gene expression of telomere-associated factors and telomerase components in different male reproductive tissues during postnatal development indicates that maintenance of telomere length would be regulated in tissue- and/or age-specific manners.

• 한국발생생물학회지:발생과생식
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• 제20권3호
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• pp.237-245
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• 2016

Direct communication between neighboring cells via gap junction in tissue is important for maintenance and regulation of its physiological functions. Each epididymal region has different composition of cell types. It is well recognized that the epididymis is a steroid hormone-responsive tissue. The present study was designed to determine the effect of estradiol benzoate (EB) or flutamide exposured at the early postnatal age on the expression of connexin (Cx) isoforms in the caudal epididymis. The EB or flutamide was subcutaneously administrated to male Spragure Dawley rat at 7 days of age, and expressional changes of Cx isoforms in the adult corpus epididymis were determined by quantitative real-time PCR. The treatment of low-dose EB resulted in decreases of Cx30.3, Cx31.1, Cx37, and Cx45 expression but caused an increase of Cx32 expression. Exposure to high-dose EB led into expressional increases of Cx31, Cx31.1, Cx32, Cx40, and Cx43, even though a decrease of Cx37 expression was found with a high-dose EB treatment. A low-dose flutamide induced increases of Cx31, Cx31.1, Cx32, and Cx43 expression but a decrease of Cx37 expression. Expression of most Cx genes were significantly increased by a high-dose flutamide, while no expressional change of Cx26 and Cx40 was detected by a high-dose flutamide. These results indicate that expression of Cx isoforms in the caudal epididymis is altered by exposure to steroidal compounds at the prepubertal age. It is suggested that a contact with environmental exogenous materials during the early postnatal period would lead to alteration of epididymal functions at the adult.

• 한국발생생물학회지:발생과생식
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• 제26권4호
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• pp.165-174
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• 2022

Three proteins [myosin heavy chain (MHC), filamin-C fragment (FIL-C), and actin 2 (ACT2)] were identified in adductor muscle from diploid and triploid Pacific oysters (Crassostrea gigas) and the relationship between the condition index (CI) and mRNA expression of these genes was investigated, together with the mRNA expression of molluscan insulin-related peptide (MIP), C. gigas insulin receptor-related receptor (CIR), and insulin-like growth factor binding protein complex acid labile subunit (IGFBP-ALS). Monthly changes in the CI were similar to the changes in the tissue weight rate in both groups. ACT2 and MHC mRNA expression was statistically higher in the triploid than the diploid, while FIL-C mRNA expression was significantly higher in the diploid (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression of the diploid oysters were all significantly higher in July than in other months (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression in the triploid oysters was high in July, but there were no significant differences (p>0.05). Changes in the expression levels of the genes investigated in this study could be used as intrinsic indicators of the annual growth, maturity, and spawning period of cultured diploid and triploid C. gigas in Tongyeong, Korea.

• Reproductive and Developmental Biology
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• 제33권1호
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• pp.19-24
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• 2009

This study was conducted to investigate the specific expression genes in the cloned bovine tissues. Donor cells, cloned tissues were analysed by RAPD-RFLP method. The results were detected three genes (CH-U7B, CH-U7M and CH-U7P) in the cloned fetus. It was found a single copy genes by southern hybridization. Sequence analysis of CH-U7M gene was shown 99% homology to a previously reported EST from a cloned bovine fetus. The putative ORF was encode a protein of hydrophobicity index 0.03. Semi-quantitative RT-PCR by using the CH-LS001 specific primer was remarkably detected in the lung tissue of cloned fetus. Further investigation of these genes may provide one of the key information to explain the early death, abnormal fetus, large off-spring and the low pregnancy rate in the production of cloned bovine.

• Reproductive and Developmental Biology
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• 제33권4호
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• pp.195-202
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• 2009

Abnormal development and fetal loss during the post-implantation period are key concerns in the production of cloned animals by somatic cell nuclear transfer (SCNT). We hypothesized that the problems in cloned porcine offspring derived from SCNT are related to interactions between the conceptus and the endometrial environment. In the present study, we investigated expression patterns in the formation of placenta-related genes (Cdx2 and GATA6) in whole in vivo normal porcine embryos (from single cell to blastocyst) and each tissue of a normal fetus at Days 25, 35 and 55 by quantitative mRNA expression analysis using real-time PCR. The expression of Cdx2 and GATA6 mRNA increased to around the blastocyst stage. These genes were gradually decreased from the peri-implantation to post-implantation stage. Moreover, we examined the expression patterns of Cdx2 and GATA6 in Day 35 normal and SCNT cloned fetuses by the same methods. And, the level of Cdx2 and GATA6 gene expression in the extraembryonic tissue of SCNT was significantly higher than that of control tissues. From the present results, it can be postulated that the aberrant expression of Cdx2 and GATA6 genes in the endometrial and extraembryonic tissues at pre- and peri-implantation stages may be closely related to the lower efficiency of animal cloning.

• 한국발생생물학회지:발생과생식
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• 제24권3호
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• pp.187-196
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• 2020

The caspase10 encodes an initiating caspase that plays an important role in the maintaining the cellular homeostasis by regulating the steps involved in the immune response and cell death. We investigated the expression of caspase10 during the different developmental stages and in olive flounder tissues. Caspase10 increased in the late stage of the formation of immune tissue, and high expression was observed in the gills, kidney, skin, and spleen. The current study analyzed the expressional changes of caspase10 in olive flounder infected with viral hemorrhagic septicemia virus (VHSV). One of the major causes of mass mortality, VHSV infection in olive flounder attributes to significant expression of caspase10 in the gills, spleen, skin, and kidneys. The results indicate a close association of caspase10 expression with the immune response to VHSV infection in olive flounder. The observations could form the basis data for exploration of other fish immune system.

• Asian-Australasian Journal of Animal Sciences
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• 제27권12호
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• pp.1684-1690
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• 2014

Estrogen and its receptors are essential hormones for normal reproductive function in males and females during developmental stage. To better understand the effect of estrogen receptor (ER) gene in yak (Bos grunniens), reverse transcription-polymerase chain reaction (PCR) was carried out to clone $ER{\alpha}$ and $ER{\beta}$ genes. Bioinformatics methods were used to analyze the evolutionary relationship between yaks and other species, and real-time PCR was performed to identify the mRNA expression of $ER{\alpha}$ and $ER{\beta}$. Sequence analysis showed that the ER open reading frames (ORFs) encoded 596 and 527 amino acid proteins. The yak $ER{\alpha}$ and $ER{\beta}$ shared 45.3% to 99.5% and 53.9% to 99.1% protein sequence identities with other species homologs, respectively. Real-time PCR analysis revealed that $ER{\alpha}$ and $ER{\beta}$ were expressed in a variety of tissues, but the expression level of $ER{\alpha}$ was higher than that of $ER{\beta}$ in all tissues, except testis. The mRNA expression of $ER{\alpha}$ was highest in the mammary gland, followed by uterus, oviduct, and ovary, and lowest in the liver, kidney, lung, testis, spleen, and heart. The $ER{\beta}$ mRNA level was highest in the ovary; intermediary in the uterus and oviduct; and lowest in the heart, liver, spleen, lung, kidney, mammary gland, and testis. The identification and tissue distribution of ER genes in yaks provides a foundation for the further study on their biological functions.