Biomedical subjects

Zuomin Zhou

Publications and source records attributed to Zuomin Zhou.

At least 19 recordsLinked to original sources

PubMed2006

NYD-SP15: a novel gene potentially involved in regulating testicular development and spermatogenesis.

By hybridizing human adult testis cDNA microarrays with human adult and embryo testis cDNA probes, we identified a novel human testis gene, NYD-SP15. NYD-SP15 expression was 3.26-fold higher in adult than in fetal testis; however, there was almost no NYD-SP15 expression in the sperm. NYD-SP15 comprises 3364 base pairs, including a 1545 bp open reading frame encoding a 514 amino acid protein possessing 89% sequence identity with the mouse testis homologous protein. NYD-SP15 is located on human chromosome 13q14.2. The deduced structure of the protein contains two dCMP_cyt_deam domains, indicating a potential functional role for zinc ion binding. The gene is expressed variably in a wide range of tissues, with high expression levels in the testis. Sequence analysis revealed that NYD-SP15 is not a highly conserved protein, with its distribution in high-level species such as vertebrates including Homo, Mus, Rattus, and Canis. The results of semiquantitative polymerase chain reaction in mouse testis representing different developmental stages indicate that NYD-SP15 expression was developmentally regulated. These results suggest the putative NYD-SP15 protein may play an important role in testicular development and spermatogenesis and may be an important factor governing male infertility.

Amino Acid Sequence↗

PubMed2006

A testis-specific and testis developmentally regulated tumor protein D52 (TPD52)-like protein TPD52L3/hD55 interacts with TPD52 family proteins.

Tumor protein D52-like proteins (TPD52) are small coiled-coil motif bearing proteins that were first identified in breast cancer. TPD52 and related proteins have been implicated in cell proliferation, apoptosis, and vesicle trafficking. To date, three human TPD52 members had been identified, named hD52 (TPD52), hD53 (TPD52L1), and hD54 (TPD52L2). The most important characteristic of the protein family is a highly conserved coiled-coil motif that is required for homo- and heteromeric interaction with other TPD52-like proteins. Herein, we identified a novel TPD52-like sequence (TPD52L3, or hD55) in human testis using cDNA microarray. Sequence analysis of the deduced protein suggests that hD55 contains a coiled-coil motif and is highly conserved compared with other TPD52-like sequences. Yeast two-hybrid and GST pull-down assays revealed that hD55 interacts with hD52, hD53, hD54, and itself. cDNA microarray detection found that hD55 was expressed at 5.6-fold higher levels in adult testis than in fetal testis. Additionally, the expression profile shows that hD55 is testis-specific, indicating a potential role for hD55 in testis development and spermatogenesis.

Aging↗

PubMed2006

Defective expression of Galpha12 in the testes of azoospermia patients and in the spermatozoa with low motility.

Antibody to the Galpha12-subunit of guanine nucleotide regulatory proteins was used to determine whether the Galpha12 is present in adult human spermatogenic cells and to determine its role in dyszoospermia. Immunoblots from testes and spermatozoa demonstrated the presence of Galpha12 in the samples. Immunohistochemical analyses of testes found that Galpha12 was expressed in the cytoplasm of Leydig cells and was expressed in spermatids from the elongating Sb phase to mature sperm. Indirect immunofluorescence of human spermatozoa revealed the presence of Galpha12 in the neck region and the midpiece of the sperm. Galpha12 in spermatids and spermatozoa partially co-localized with F-actin and alpha-tubulin. Immunohistochemical analyses of tissues from three patients with non-obstructive azoospermia showed abnormal expression of Galpha12 in more than 45% of spermatids. Furthermore, Western blots and indirect immunofluorescence found defective expression of Galpha12 in low-motility spermatozoa with midpieces that were bent on themselves. Therefore, it suggests that Galpha12 plays a role in polarity and tail formation as spermatids mature. Furthermore, Galpha12 may be a candidate protein responsible for azoospermia caused by spermatogenic disturbance or midpiece deformities.

Actins↗

PubMed2006

Human testis specific protein 1 expression in human spermatogenesis and involvement in the pathogenesis of male infertility.

Human testis specific protein 1 (TPX1) exists in the cytomembrane and cytoplasm of spermatogenic cells from pachytene spermatocytes to elongated spermtids, including pachytene spermatocytes, round spermtids and elongated spermtids. It is localized in the connecting piece, the flagellum, and the acrosome of mature human spermatozoa. The protein level and localization of TPX1 were altered in patients with spermatogenic arrest and in infertile men with oligoasthenoteratospermia syndrome.

Adult↗

PubMed2006

Involvement of ALF in human spermatogenesis and male infertility.

We conducted this study to explore functions of TF II Aalpha/beta-like factor (ALF) during human spermatogenesis, and the relationship of its expression levels with male infertility. The RT-PCR and Western blot analyses illustrated that ALF was highly expressed in adult testis. Immunohistochemistry and immunoflurescence showed that ALF is located in the spermatid nuclei and in the annulus of spermatozoa. Further, to reveal whether ALF is related to male infertility, we performed the same experiments in infertility patients. The changes in the expression levels of ALF in the male infertility samples lead us to believe that ALF may function in spermatogenesis, especially in spermiogenesis. We also detected the ALF DNA methylation level by real-time methylation-specific PCR (MSP) both in testes of adult, fetal and infertile patient. The differential expression level of ALF gene in different types of testes was regulated by DNA methylation. Our research identified ALF as a human spermatogenesis related gene, the abnormal expression of ALF might be the partial cause for human infertility.

Gene Expression Profiling↗

PubMed2006

Cloning and characterization of a novel sperm tail protein, NYD-SP28.

In this study, a gene coding a novel human sperm tail protein named NYD-SP28 was cloned and characterized using a complementary DNA (cDNA) microarray. Its expression was 3.5 times higher in human testis than in fetal testis, and very high in human spermatozoa. The full length of NYD-SP28 cDNA was 1798 bp and encoded a 484-amino-acid protein. Motif analysis revealed that the protein contained a cluster of phosphorylation sites, N-glycosylation sites and N-myristoylation sites. Immunohistochemical analysis of normal human testes showed that NYD-SP28 was expressed in the cytoplasm of spermatogenic cells but not in interstitial cells. The EGFP-NYD-SP28 fusion protein was also localized in the cytoplasm of transfected 7721 cells. In human spermatozoa, NYD-SP28 immunoreactivity was detected in entire sperm tail. Using the two-dimensional (2-D) gel electrophoresis and immunoblotting technique, NYD-SP28 was found to be post-translationally modified during sperm capacitation. In conclusion, these results suggest that NYD-SP28 is a new human sperm tail protein and might play an important role during sperm capacitation.

Adult↗

PubMed2005

Molecular cloning, identification and characteristics of a novel isoform of carbamyl phosphate synthetase I in human testis.

A gene coding a novel isoform of carbamyl phosphate synthetase I (CPS1) was cloned from a human testicular library. As shown by cDNA microarray hybridization, this gene was expressed at a higher level in human adult testes than in fetal testes. The full length of its cDNA was 3831 bp, with a 3149 bp open reading frame, encoding a 1050-amino-acid protein. The cDNA sequence was deposited in the GenBank (AY317138). Sequence analysis showed that it was homologous to the human CPS1 gene. The putative protein contained functional domains composing the intact large subunit of carbamoyl phosphate synthetase, thus indicated it has the capability of arginine biosynthesis. A multiple tissue expression profile showed high expression of this gene in human testis, suggesting the novel alternative splicing form of CPS1 may be correlated with human spermatogenesis.

Adult↗

PubMed2005

Identification of a novel substrate for tyrosine kinase in human testes.

Identification of genes specifically expressed in adult and fetal testes is important in furthering our understanding of testis development and function. In this study, a novel human specific transcript, designated NYD-SP10, was identified by hybridization of adult and fetal human testis probes with a human cDNA microarray. NYD-SP10 (GenBank Accession no. AF332192) was expressed at a higher level in adult testes than in fetal testes. Bioinformatic analysis illustrated NYD-SP10 is a splice variant of regulatory factor X4, while it is lack of RFX DNA-binding domain which binds to the X box of MHC class II genes. Most telling of all, NYD-SP10 uniquely possesses a tyrosine kinase phosphorylation site. Expression profiles demonstrated NYD-SP10 is specifically and strongly expressed in human adult testes and ejaculated spermatozoa. Taken together, these results suggest that NYD-SP10 is a novel testis-specific gene and it is possibly a novel substrate for tyrosine kinase and involved in the function of human spermatozoa, especially during capacitation.

Adult↗

PubMed2005

Molecular cloning and expression of a novel alternative splice variant of BRDT gene.

In this study, a human adult testis cDNA microarray was constructed and hybridized with (33)P-labeled human adult testis, embryo testis and sperm cDNA probes, respectively. A novel alternative splice variant of BRDT gene, named BRDT-NY, presumably involved in testicular function was cloned. It was expressed 3.96-fold more in human adult than embryo testis and also expressed in human spermatozoa. Similarly, RT-PCR revealed a differential expression pattern of this gene in human adult testes and fetal testes. The full length of BRDT-NY was 3438 bp and contained a 2883 bp open reading frame, encoding a 960-amino-acid protein. Sequence analysis showed that it has two bromodomains in N-terminal of the protein. Multiple tissue RT-PCR results showed that BRDT-NY was exclusively expressed in testis. mRNA expression of BRDT-NY gene was deleted in some azoospermic patients' testes. These experiments suggested that BRDT-NY gene may have an important role in the process of spermatogenesis and may be correlated with male infertility.

Alternative Splicing↗

PubMed2005

Cloning and characterization of a novel intronless lactate dehydrogenase gene in human testis.

Using cDNA microarray hybridization from a human testicular cDNA library, one gene named lactate dehydrogenase A-like gene (LDHL, also known as LDHL6B) was cloned. LDHL exhibited 3.8-fold difference at expression level between adult and fetal human testes. The full cDNA length of LDHL is 1680 bp and had a 1145 bp open reading frame, which encoded a 41.9 kDa protein of 381 amino acids. Sequence analysis showed that LDHL harbors all the domains (one lactate/malate dehydrogenase, NAD binding domain and one lactate/malate dehydrogenase, alpha/beta C-terminal domain) in lactate dehydrogenase gene family. Blasting human genome database localized LDHL to human chromosome 15q22.2 and it was an intronless gene. Results of multiple-tissue PCR and real-time PCR showed that LDHL expressed mainly in testis and its mRNA abundance was testis development-related. In summary, LDHL is believed to be involved in testis development and spermatogenesis.

Amino Acid Sequence↗

PubMed2005

SMYD3-NY, a novel SMYD3 mRNA transcript variant, may have a role in human spermatogenesis.

Identification of genes specifically expressed in adult and fetal testis is important to further our understanding of testis development and function. In this study, a novel SMYD3 transcript variant, termed SMYD3-NY (GenBank Accession No. AY186742), was identified by hybridization of adult and fetal human testis cDNA probes with a human cDNA microarray. SMYD3-NY transcript was expressed at 2.3-fold higher levels in adult human testis than in fetal testis, with a low expression level in human spermatozoa. Bioinformatical analysis showed that SMYD3-NY protein has the SET domain that is involved in histone methyltransferase (HTMase) activity. Southern blotting showed that SMYD3-NY is distributed in several tissues, including testis. In summary, SMYD3-NY is a novel transcript variant of the SMYD3 gene, and SMYD3-NY protein may influence transcriptional regulation during spermatogenesis via HTMase activity.

Adult↗

PubMed2004

A spermatogenesis-related gene expression profile in human spermatozoa and its potential clinical applications.

Spermatogenesis is an essential stage in the human reproductive process. In a previously study aiming to determine which genes might be involved in spermatogenesis, we compared the gene expression profiles of adult and fetal testes by hybridizing cDNA probes prepared from adult and fetal testes to membranes dotted with gene clones derived from a commercial human testis library. We identified 266 differentially expressed genes that showed higher expression levels in adult testes, indicating their potential roles in spermatogenesis. In the present study, we applied the same cDNA microarray technique to the analysis of gene expression in the spermatozoa of normal fertile men and found 149 genes that were expressed at higher levels in adult testis. A further study of five sperm motility-related genes selected from this profile by real-time PCR revealed that there was significant difference in the expression levels of two genes ( TPX-1, testis-specific protein 1 and LDHC, lactate dehydrogenase C, transcript variant 1) between normal ( n=29) and motility impaired ( n=24) semen samples, indicating that these genes are involved in sperm function. Our results demonstrated that spermatogenesis-related gene profiling could help to assess sperm quality in humans, and further study of these genes will help us to elucidate the mechanisms involved in spermatogenesis and diseases relating to human infertility.

A Kinase Anchor Proteins↗

PubMed2004

Expression of a novel DnaJA1 alternative splicing in human testis and sperm.

Using cDNA microarray hybridization from a human testicular cDNA library, one gene exhibiting threefold difference at expression level between adult and embryo human testes was named nDnaJA1 (a new alternative isoform of human DnaJA1 which was also named HDJ2/Hsdj/dj2, a human HSP40 homologue), which was believed to be involved in testis development and spermatogenesis. Multiple tissue polymerase chain reaction (PCR) results showed that nDnaJA1 expressed highly in testis and lung but low in thymus, prostate, colon and liver. The results of the other members of DnaJA1 family (GenBank accession numbers: D13388 and BC008182) showed that they were widely expressed. D13388 and BC008182 were highly expressed in sperm while nDnaJA1 was faintly expressed by reverse transcriptase PCR. Protein motif analysis of nDnaJA1 sequence revealed motifs of DnaJ. nDnaJA1 was considered as type I DnaJ like the other members of DnaJA1 family.

Alternative Splicing↗

PubMed2004

Cloning and expression of a novel CREB mRNA splice variant in human testis.

Identification of genes specifically expressed in adult and fetal testis is important in furthering our understanding of testis development and function. In this study, a novel human transcript, designated human testis cAMP-responsive element-binding protein (htCREB), was identified by hybridization of adult and fetal human testis cDNA probes with a human cDNA microarray containing 9216 clones. The htCREB transcript (GenBank Accession no. AY347527) was expressed at 2.35-fold higher levels in adult human testes than in fetal testes. Sequence and ntBLAST analyses against the human genome database indicated that htCREB was a novel splice variant of human CREB. RT-PCR-based tissue distribution experiments demonstrated that the htCREB transcript was highly expressed in adult human testis and in healthy sperm, but not in testes from patients with Sertoli cell-only syndrome. Taken together, these results suggest that the htCREB transcript is chiefly expressed in germ cells and is most likely involved in spermatogenesis.

Alternative Splicing↗

PubMed2004

The molecular characteristics of polycystic ovary syndrome (PCOS) ovary defined by human ovary cDNA microarray.

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders; it is characterized by polycystic ovaries, hyperandrogenism and chronic anovulation. To obtain a global view of those genes that might be involved in the development of this complex clinical disorder, we used recently developed cDNA microarray technology to compare differential gene expressions between normal human ovary and ovaries from PCOS patients. A total of 9216 clones randomly selected from a commercial human ovary cDNA library were screened. Among them, 290 clones showed differential expressions, including 119 known genes and 100 known or unknown expressed sequence tags (ESTs). Among 119 known genes, 88 were upregulated and 31 downregulated in the PCOS ovary, as compared with normal human ovary. These differentially expressed genes are involved in various biologic functions, such as cell division/apoptosis, regulation of gene expression and metabolism, reflecting the complexity of clinical manifestations of PCOS. The molecular characteristics established from our study will further our understanding of the pathogenesis of PCOS and help us to identify new targets for further studies and for the development of new therapeutic interventions.

Base Sequence↗

PubMed2004

[Phospholipase C and reproduction].

Phospholipase C (PLC) is a key enzyme in phosphatidyl inositol turnover during signal transduction. The 12 mammalian PLC isozymes identified to date can be divided into five subtypes, beta-type, gamma-type, delta-type, epsilon-type and zeta-type, with extensive difference in structure, regulatory mechanism and tissue distribution. PLC plays important roles not only in sperm acrosome reaction but also in egg activation. The present studies are reviewed on the structure, regulation and function of PLC, especially its function in male reproduction, including triggering Ca2+ oscillations in eggs to activate the eggs and helping embryo development. And the prospect of the clinical application of PLC is discussed.

Acrosome Reaction↗

PubMed2003

Rescue of defective pancreatic secretion in cystic-fibrosis cells by suppression of a novel isoform of phospholipase C.

BACKGROUND: Cystic fibrosis is caused by mutations in the gene encoding an ion-transport protein, the cystic-fibrosis transmembrane conductance regulator (CFTR). Defective secretion of anions is the primary cause of many of the clinical manifestations of cystic fibrosis, including pancreatic insufficiency. We aimed to identify a molecular mechanism from which a new method to circumvent defective pancreatic secretion could be derived. METHODS: Multiple-human-tissue RT-PCR and semiquantitative RT-PCR analyses were used to examine gene expression. An antisense technique was used in conjunction with radioimmunoassay, Fura-2 spectrofluorometry, immunohistochemistry, and the short-circuit current technique (Ussing chamber) for elucidation of gene function and its application in rescuing defective pancreatic secretion. FINDINGS: We cloned a newly identified gene, NYD-SP27, which has structural similarity to an isoform of phospholipase C. NYD-SP27 was expressed endogenously in human pancreatic-duct cells and upregulated in cystic fibrosis. Suppression of NYD-SP27, by transfection of its antisense into human cystic-fibrosis pancreatic-duct cells, resulted in augmentation of phospholipase-C-coupled calcium-ion release and protein kinase C activity, improvement in the amount of mutated CFTR reaching the plasma membrane, and restoration of cAMP-activated pancreatic anion secretion. INTERPRETATION: NYD-SP27 exerts an inhibitory effect on phospholipase-C-coupled processes that depend on calcium ions and protein kinase C, including CFTR trafficking and function. Its upregulation in pancreatic-duct cells may reveal a previously unsuspected defect in cystic fibrosis contributing to pancreatic insufficiency, and thus represents a new target for pharmacological intervention in cystic fibrosis.

Antisense Elements (Genetics)↗

PubMed2003

Identification and characterization of a novel human testis-specific Golgi protein, NYD-SP12.

A novel human testis-specific gene, NYD-SP12, was identified by hybridizing human adult or fetal testes cDNA samples with a human cDNA microarray containing 9216 clones. mRNA expression level of NYD-SP12 was 30-fold higher in human adult testes than fetal testes. Similarly, semi-quantitative RT-PCR revealed a differential expression pattern of an NYD-SP12 homologous gene in mouse adult and infant testes. PCR and hybridization analysis of NYD-SP12 mRNA from multiple human tissues indicated the expression of NYD-SP12 exclusively in the testis. In-situ hybridization revealed that the expression of this gene was confined to spermatogenic epithelium and was not found in interstitial cells. NYD-SP12 transcript was not detected in patients with spermatogenic arrest and Sertoli cell-only syndrome. NYD-SP12 cDNA (GenBank accession number: AF345909) consisted of 2070 bp. The predicted 1707 bp open-reading fragment encoded a 569 amino acid protein that was 77% identical to a mouse homologue. Furthermore, computerized SMART and Motif analysis revealed that the protein contained a Structural Classification Of Proteins (SCOP) domain in the C-terminus and a cluster of phosphorylation sites for PKC, CK and cAMP/cGMP-dependent protein kinase. Interestingly, the EGFP-NYD-SP12 fusion protein was localized to the Golgi apparatus. In conclusion, the results suggest that NYD-SP12 is involved in spermatogenesis, and that NYD-SP12-encoded protein might function in the Golgi apparatus.

Amino Acid Sequence↗