• Title/Summary/Keyword: Testis-specific expression

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Expression of MAGE in Gastric Cancer Tissues (위암조직에서의 MAGE 유전자 발현)

  • Choi, Jae-Young;Lee, Sang-Ho
    • Journal of Gastric Cancer
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    • v.5 no.3 s.19
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    • pp.180-185
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    • 2005
  • Purpose: Among tumor-associated antigens, MAGE (melanoma antigen) was named as cancer/testis specific antigens because they are detected exclusively in the testis or cancer cells, including gastric carcinomas. Due to the elicitation of autoimmunitiy to tumors by these antigens either in vitro or in vivo and their tumor specificity, these antigens, thus, appear to be potential targets for tumor-specific immunotherapy. Materials and Methods: The fresh tumor tissue and normal gastric tissue samples were obtained from resected surgical specimens in 53 patients with gastric carcinomas. From the obtained cells, total cellular mRNA was extracted, and RT-PCR and nested PCR were run in 30 and 35 cycles respectively, with two different kinds of primers specially designed to detect six subtypes of MAGE DNA simultaneously. Results: In the 53 normal tissue, there was no expression of MAGE, but in the 53 cancer tissues, MAGE was expressed in 13 tissues (24.5%). Our data did not exhibit any correlation with the expression of the MAGE gene and clinicopathological factors. Conclusion: In our data, since 24.5% of gastric cancer tissues expressed MAGE, it should become possible to immunize a significant proportion of patients with advanced gastric carcinomas against the antigens encoded by these genes, provided that more antigenic peptides encoded by the genes of the MAGE family can be identified in the near future. (J Korean Gastric Cancer Assoc 2005;5:180-185)

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Nam and Kim #1 Tissue-specific expression of pFV4CAT in transgenic mud loach (Misgurnus mizolepis) germ line (미꾸라지(Misgurnus mizolepis)에서 pFV4CAT 의 조직 특이적 발현)

  • Nam, Yoon-Kwon;Kim, Dong-Soo
    • Korean Journal of Ichthyology
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    • v.9 no.1
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    • pp.91-98
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    • 1997
  • The transgene, pFV4CAT, containing CAT reporter gene regulated by carp $\beta$-actin promoter, was expressed in independent transgenic mud loach germ lines, determined by reverse transcriptase-PCR (RT-PCR) and enzyme-linked immunosorbant assay (ELISA). Expression of the transmitted transgene was found to be tissue-specific in F1 and F2 generations. Tissue specificity of the expression was dependent on each transgenic line with reproducible patterns. Liver and spleen did express the transgene more frequently than other tissues tested, and muscle and heart revealed the higher amount of CAT than other tissues, while testes showed the lowest expression level. The highest level of CAT expression in muscle from a transgenic F1 line was corresponding to 68-fold compared to the basal levels of controls.

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Molecular Cloning and Bioinformatic Analysis of SPATA4 Gene

  • Liu, Shang-Feng;Ai, Chao;Ge, Zhong-Qi;Liu, Hai-Luo;Liu, Bo-Wen;He, Shan;Wang, Zhao
    • BMB Reports
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    • v.38 no.6
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    • pp.739-747
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    • 2005
  • Full-length cDNA sequences of four novel SPATA4 genes in chimpanzee, cow, chicken and ascidian were identified by bioinformatic analysis using mouse or human SPATA4 cDNA fragment as electronic probe. All these genes have 6 exons and have similar protein molecular weight and do not localize in sex chromosome. The mouse SPATA4 sequence is identified as significantly changed in cryptorchidism, which shares no significant homology with any known protein in swissprot databases except for the homologous genes in various vertebrates. Our searching results showed that all SPATA4 proteins have a putative conserved domain DUF1042. The percentages of putative SPATA4 protein sequence identity ranging from 30% to 99%. The high similarity was also found in 1 kb promoter regions of human, mouse and rat SPATA4 gene. The similarities of the sequences upstream of SPATA4 promoter also have a high proportion. The results of searching SymAtlas (http://symatlas.gnf.org/SymAtlas/) showed that human SPATA4 has a high expression in testis, especially in testis interstitial, leydig cell, seminiferous tubule and germ cell. Mouse SPATA4 was observed exclusively in adult mouse testis and almost no signal was detected in other tissues. The pI values of the protein are negative, ranging from 9.44 to 10.15. The subcellular location of the protein is usually in the nucleus. And the signal peptide possibilities for SPATA4 are always zero. Using the SNPs data in NCBI, we found 33 SNPs in human SPATA4 gene genomic DNA region, with the distribution of 29 SNPs in the introns. CpG island searching gives the data about CpG island, which shows that the regions of the CpG island have a high similarity with each other, though the length of the CpG island is different from each other.This research is a fundamental work in the fields of the bioinformational analysis, and also put forward a new way for the bioinformatic analysis of other genes.

Analysis of Pregnancy-Associated Plasma Protein-A (PAPP-A) in Porcine Neonatal Testis (미성숙 돼지 정소 내에서의 pregnancy-associated plasma protein-A 특성 분석)

  • Lee, W.Y.;Cho, K.H.;Yeo, J.M.;Shin, Y.K.;Park, J.K.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.1
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    • pp.5-13
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    • 2020
  • The identify of biomarkers in living tissues is useful to understand the characteristics and functions of the cells. Proteins such as protein gene product 9.5, promyelocytic leukemia zinc finger, NANOG, and stage-specific embryonic antigen-1 have been identified as markers for porcine undifferentiated spermatogonia. In this study, the expression of insulin-like growth factor binding proteins (IGFBPs), a newly discovered porcine spermatogonia marker and pregnancy-associated plasma protein-A (PAPP-A), a protein regulator of IGFBPs, were characterized in 5-day-old porcine testis. To analyze the function of IGFBPs, RT-PCR was performed. IGFBP 2, 3, 4, and 6 were detected in porcine spermatogonia and PAPP-A was detected in basement regions in 5day old porcine seminiferous tubules. PAPP-A was not expressed in spermatogonia, but it was expressed in Sertoli cells. These results suggest that the expression of PAPP-A protein in Sertoli cells may regulate the development and differentiation of testicular cells through the IGF axis in porcine neonatal testis.

Effects of Extracellular Matrix Protein-derived Signaling on the Maintenance of the Undifferentiated State of Spermatogonial Stem Cells from Porcine Neonatal Testis

  • Park, Min Hee;Park, Ji Eun;Kim, Min Seong;Lee, Kwon Young;Hwang, Jae Yeon;Yun, Jung Im;Choi, Jung Hoon;Lee, Eunsong;Lee, Seung Tae
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1398-1406
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    • 2016
  • In general, the seminiferous tubule basement membrane (STBM), comprising laminin, collagen IV, perlecan, and entactin, plays an important role in self-renewal and spermatogenesis of spermatogonial stem cells (SSCs) in the testis. However, among the diverse extracellular matrix (ECM) proteins constituting the STBM, the mechanism by which each regulates SSC fate has yet to be revealed. Accordingly, we investigated the effects of various ECM proteins on the maintenance of the undifferentiated state of SSCs in pigs. First, an extracellular signaling-free culture system was optimized, and alkaline phosphatase (AP) activity and transcriptional regulation of SSC-specific genes were analyzed in porcine SSCs (pSSCs) cultured for 1, 3, and 5 days on non-, laminin- and collagen IV-coated Petri dishes in the optimized culture system. The microenvironment consisting of glial cell-derived neurotrophic factor (GDNF)-supplemented mouse embryonic stem cell culture medium (mESCCM) (GDNF-mESCCM) demonstrated the highest efficiency in the maintenance of AP activity. Moreover, under the established extracellular signaling-free microenvironment, effective maintenance of AP activity and SSC-specific gene expression was detected in pSSCs experiencing laminin-derived signaling. From these results, we believe that laminin can serve as an extracellular niche factor required for the in vitro maintenance of undifferentiated pSSCs in the establishment of the pSSC culture system.

Expressions of Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptor Gene in the Rat Uterus (흰쥐 자궁에서 Pituitary Adenylate Cyclase-Activating Polypeptide와 수용체 유전자의 발현)

  • 이성호
    • Development and Reproduction
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    • v.2 no.1
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    • pp.21-27
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    • 1998
  • The present study was performed to analyze the gene expressions of pituitary adenylate cyclase-activating polypeptide(PACAP) and its receptor in the rat uterus, a candidate for novel extrahypothalamic source and target. The PACAP cDNA fragments corresponding to the common exon region which is found in both the rat hypothalamus and testis were produced from all tissue samples including the rat uterus by reverse transcriptionpolymerase chain reaction (RT-PCR). No PCR product was amplified from the rat hypothalamic, pituitary, ovarian and uterine samples when the 5' primer corresponding to the testis-specific exon 1 region was used, while the predicted size of product was detected from the testis sample. RT-PCR using the uterine RNA and specific primers for the PACAP receptor yielded products with predicted sizes. Transcripts for the rat uterine PACAP receptor were identified as type I isoforms with hip-hop and hip- or hop-type inserts. After pregnant mare's serum gonadotropin (15 IU) treatment of immature rats (day 25), the level of PACAP mRNA was increased in 24 h and 48 h group, and was declined to the lowest in 72 h group. The present study shows the presence of transcripts for PACAP and its receptor isoform in the rat uterus. These finding ssuggest that the uterine PACAP ight act as a novel autocrine and/or paracrine factor via its specific receptors on the reglulation of rat uterine function and physiology during the reproductive cycle.

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Molecular Characterization and Expression Pattern of Gene IGFBP-5 in the Cashmere Goat (Capra hircus)

  • Wang, X.J.;Shi, J.J.;Yang, J.F.;Liang, Y.;Wang, Y.F.;Wu, M.L.;Li, S.Y.;Guo, X.D.;Wang, Z.G.;Liu, D.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.606-612
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    • 2012
  • Insulin-like growth factor-binding protein-5 (IGFBP-5) is one of the six members of IGFBP family, important for cell growth, apoptosis and other IGF-stimulated signaling pathways. In order to explore the significance of IGFBP-5 in cells of the Inner Mongolian Cashmere goat (Capra hircus), IGFBP-5 gene complementary DNA (cDNA) was amplified by reverse transcription polymerase chain reaction (RT-PCR) from the animal's fetal fibroblasts and tissue-specific expression analysis was performed by semi-quantitative RT-PCR. The gene is 816 base pairs (bp) in length and includes the complete open reading frame, encoding 271 amino acids (GenBank accession number JF720883). The full cDNA nucleotide sequence has a 99% identity with sheep, 98% with cattle and 95% with human. The amino acids sequence shares identity with 99%, 99% and 99%, respectively. The bioinformatics analysis showed that IGFBP-5 has an insulin growth factor-binding protein homologues (IB) domain and a thyroglobulin type-1 (TY) domain, four protein kinase C phosphorylation sites, five casein kinase II phosphorylation sites, three prenyl group binding sites (CaaX box). The IGFBP-5 gene was expressed in all the tested tissues including testis, brain, liver, lung, mammary gland, spleen, and kidney, suggesting that IGFBP-5 plays an important role in goat cells.

Gain of New Exons and Promoters by Lineage-Specific Transposable Elements-Integration and Conservation Event on CHRM3 Gene

  • Huh, Jae-Won;Kim, Young-Hyun;Lee, Sang-Rae;Kim, Hyoungwoo;Kim, Dae-Soo;Kim, Heui-Soo;Kang, Han-Seok;Chang, Kyu-Tae
    • Molecules and Cells
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    • v.28 no.2
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    • pp.111-117
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    • 2009
  • The CHRM3 gene is a member of the muscarinic acetylcholine receptor family that plays important roles in the regulation of fundamental physiological functions. The evolutionary mechanism of exon-acquisition and alternative splicing of the CHRM3 gene in relation to transposable elements (TEs) were analyzed using experimental approaches and in silico analysis. Five different transcript variants (T1, T2, T3, T3-1, and T4) derived from three distinct promoter regions (T1: L1HS, T2, T4: original, T3, T3-1: THE1C) were identified. A placenta (T1) and testis (T3 and T3-1)-dominated expression pattern appeared to be controlled by different TEs (L1HS and THE1C) that were integrated into the common ancestor genome during primate evolution. Remarkably, the T1 transcript was formed by the integration event of the human specific L1HS element. Among the 12 different brain regions, the brain stem, olfactory region, and cerebellum showed decreased expression patterns. Evolutionary analysis of splicing sites and alternative splicing suggested that the exon-acquisition event was determined by a selection and conservation mechanism. Furthermore, continuous integration events of transposable elements could produce lineage specific alternative transcripts by providing novel promoters and splicing sites. Taken together, exon-acquisition and alternative splicing events of CHRM3 genes were shown to have occurred through the continuous integration of transposable elements following conservation.

The cancer/testis antigen CAGE induces MMP-2 through the activation of NF-κB and AP-1

  • Kim, Young-Mi;Jeoung, Doo-Il
    • BMB Reports
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    • v.42 no.11
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    • pp.758-763
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    • 2009
  • Cancer-associated antigen (CAGE) induces the expression of matrix metalloproteinase-2 (MMP-2) by activating Akt, which in turn interacts with inhibitory kappa kinase $\beta$ ($I{\kappa}K{\beta}$) to activate nuclear factor ${\kappa}B$ (NF-${\kappa}B$). Akt and p38 mitogen activated protein kinase (p38 MAPK) are necessary for CAGE-mediated induction of the AP-1 subunit JunB, whereas extracellular regulated kinase (ERK) is necessary for the induction of fos-related antigen-1 (Fra-1). Induction of MMP-2 by CAGE requires activator of protein-1 (AP-1) to be bound. Specific binding of JunB to MMP-2 promoter sequences was shown by chromatin immunoprecipitation (ChIP) analysis.

Cell- and Stage-Specific Expression of the Murine nm23-M5 Gene during Late Spermatogenesis and Spermiogenesis

  • Hwang Gyu-Chan;Ok Do-Won;Lee Mi-Suk;Kim Jin-Hoe
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.5-5
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    • 2002
  • Nucleoside diphosphate kinases (NDPKs) are conserved through evolution and have been shown to be involved in various biological phenomena. By functional screening in yeast, we identified a new member of the NDPK family, ㎚23-M5, which encodes a 211-amino acid protein with 86% identify to the human homolog, ㎚23-H5. Northern blot analysis reveals that ㎚23-M5 encodes two transcripts of 0.8 and 0.7 kb, which are highly and specifically expressed in adult testis. Reverse transcriptase polymerase chain reaction analysis shows that nm23-M5 first appears in pachytene spermatocytes and increase chain reaction in abundance through subsequent stages. (omitted)

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