ABSTRACT
<p><b>AIM</b>To identify and characterize a novel gene with potential roles in testis development and spermatogenesis.</p><p><b>METHODS</b>A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature.</p><p><b>RESULTS</b>A novel testis-specific gene, NYD-SP5, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved Ile and Gln residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes.</p><p><b>CONCLUSION</b>NYD-SP5 is a newly found testis-specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.</p>
Subject(s)
Animals , Male , Mice , Amino Acid Sequence , Base Sequence , DNA, Complementary , In Situ Hybridization , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Proteins , Chemistry , Genetics , Physiology , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Spermatogenesis , Genetics , Testis , MetabolismABSTRACT
<p><b>AIM</b>To identify a novel isoform of adaptin 2 beta subunit (named Ap2beta-NY) and to investigate its relationship with testicular development and spermatogenesis.</p><p><b>METHODS</b>Using a human testis cDNA microarray, a clone (Ap2beta-NY), which was strongly expressed in adult testes but weakly expressed in embryo testes, was sequenced and analyzed. Using polymerase chain reaction (PCR), the tissue distribution and expression time pattern of Ap2beta-NY were determined.</p><p><b>RESULTS</b>Ap2beta-NY was identified and has been deposited in the GenBank (AY341427). The expression level of Ap2beta-NY in the adult testis was about 3-fold higher than that in the embryo testis. PCR analysis using multi-tissue cDNA indicated that Ap2beta-NY was highly expressed in the testis, spleen, thymus, prostate, ovary, blood leukocyte and brain, but not in the heart, placenta, lung, liver, skeletal muscle, kidney and pancreas. In addition, Ap2beta-NY was variably expressed in the testes of patients with spermatogenesis-disturbance and spermatogenesis-arrest but not expressed in those of Sertoli-cell-only syndrome, which implied that, in the testis, Ap2beta-NY was restrictively expressed in germ cells.</p><p><b>CONCLUSION</b>Ap2beta-NY is an isoform of Ap2beta and may be involved in regulating the process of spermatogenesis and testis development.</p>
Subject(s)
Humans , Male , Adaptor Protein Complex beta Subunits , Chemistry , Genetics , Amino Acid Sequence , Base Sequence , DNA, Complementary , Molecular Sequence Data , Nucleic Acid Hybridization , Oligonucleotide Array Sequence Analysis , Polymerase Chain Reaction , RNA Splicing , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Spermatogenesis , Testis , MetabolismABSTRACT
<p><b>AIM</b>To identify genes related to the human testis development by substrate hybridization technique.</p><p><b>METHODS</b>A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions. The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database. Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to determine the tissue expression profile of cul-3b.</p><p><b>RESULTS</b>Cul-3b, a novel CUL-3 transcript variant, was identified. The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones. Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites IRESes in the 5'-UTR. These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances. Additionally, cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR.</p><p><b>CONCLUSION</b>Cul-3b is a novel transcript variant of CUL-3, which may be important not only for the development of human testis but also for that of other organs.</p>
Subject(s)
Humans , Base Sequence , Cell Cycle Proteins , Genetics , Cullin Proteins , Genetics , Molecular Sequence Data , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>AIM</b>To identify a novel alternative transcript of the novel retinal pigment epithelial cell gene (NORPEG) expressed in the human testis.</p><p><b>METHODS</b>A human testis cDNA microarray was established and hybridized with cDNA probes from human fetal testes, adult testes and human spermatozoa. Differentially expressed clones were sequenced and analyzed. One of these clones was a short transcript of NORPEG which we proceeded to analyze by RT-PCR.</p><p><b>RESULTS</b>The novel short alternative transcript of NORPEG was isolated and named sNORPEG. It was 3486 bp in length and contained a 2952-bp open reading frame, encoding a 110.4-kDa protein of 983 amino acids. Amino acid sequence analysis showed that the sNORPEG protein contains six ankyrin repeats and two coiled-coil domains. It shares a high homology with the NORPEG and ankycorbin proteins in both its sequence and motifs. Blasting the human genome database localized sNORPEG to human chromosome 5p13.2-13.3. Expression profiles showed that sNORPEG was expressed in human fetal testes, adult testes and spermatozoa. Moreover, sNORPEG was found to be ubiquitously expressed in human tissues.</p><p><b>CONCLUSION</b>sNORPEG is expressed in different developmental stages of the testis and encodes a protein that may have roles in human testis development and spermatogenesis.</p>
Subject(s)
Humans , Male , Alternative Splicing , Amino Acid Sequence , Base Sequence , Cytoskeletal Proteins , Genetics , DNA, Complementary , Gene Expression Profiling , Molecular Sequence Data , Open Reading Frames , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Testis , Metabolism , Transcription Factors , GeneticsABSTRACT
<p><b>AIM</b>To identify the genes specifically expressed in human adult and fetal testes and spermatozoa.</p><p><b>METHODS</b>A human testis cDNA microarray was established. Then mRNAs of human adult and fetal testis and spermatozoa were purified and probes were prepared by a reverse transcription reaction with mRNA as the template. The microarray was hybridized with probes of adult and fetal testes and spermatozoa. The nucleic acid sequences of differentially expressed genes were determined and homologies were searched in the databases of GenBank.</p><p><b>RESULTS</b>A novel human testis-specific gene, PKH-T, was identified by hybridizing adult and fetal testis and spermatozoa probes with a human testis cDNA microarray. The cDNA of PKH-T was 1 069 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AY303972) and PKH-T was also determined as Interim GenSymbol (Unigene, HS.38041). PKH-T contained most PKH conserved motif. The 239 amino acid sequences deduced from the 719 bp open reading frame (ORF) had a homology with the gene PKH (U89606). PKH-T was specifically and strongly expressed in the testis. Comparison of the differential expressions of PKH and PKH-T in testes of different developmental stages indicated that PKH-T was expressed in the adult testis and spermatozoa, while PKH, in the adult, fetal and aged testes. PKH-T had no expression in the testis of Sertoli cell only and partially spermatogenic arrest patients.</p><p><b>CONCLUSION</b>PKH-T is a gene highly expressed in adult human testis and spermatozoa. It may play an important role in spermatogenesis and could be related to male infertility.</p>
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Amino Acid Sequence , DNA, Complementary , Genetics , Gene Expression Regulation, Enzymologic , Genetics , Infertility, Male , Genetics , Isoenzymes , Genetics , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Pyridoxal Kinase , Genetics , RNA Splicing , Genetics , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sertoli Cells , Metabolism , Spermatogenesis , Genetics , Spermatozoa , Metabolism , Testis , Embryology , Tissue DistributionABSTRACT
This article reviews recent advances in apoptosis on the pathway inducing cancer cell to death, including Bcl-2 family pathway, NF-kappa B pathway, P13K/Akt pathway, Rb gene and p53 gene, especially the targets of anticancer drug in these pathways. It could be useful for the anticancer drug design and estimate. Furthermore, those cancer/testis antigen gene products are potential targets for antigen-specific immunotherapy of carcinoma.