Full-length cDNA, molecular characterization and physical mapping of five genes from a porcine fetal cDNA library.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to Z M Zhu.
Explore the source record for details and available documents.
The proteasome subunit, beta type 10 (PSMB10) gene regulated by interferon-gamma is a core part of the 26S proteasome complex, which is an important protein degrading system. Isolation and characterization of swine PSMB10 revealed a conserved structure with other mammalian PSMB10 genes. An A/G nucleotide polymorphism in PSMB10 intron 2 and a C/T single nucleotide polymorphism in exon 5 were detected by polymerase chain reaction restriction fragment length polymorphism. The allele frequencies were significantly different among Tongcheng, Landrace, Large White and Duroc. The porcine PSMB10 was mapped by somatic cell hybrid panel and radiation hybrid mapping on SSC6p14-p15, which is in good agreement with human-pig comparative maps.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Carbonic anhydrase 3 (CA3) is a member of the carbonic anhydrase family, which plays an important role in various cell processes. In this paper, molecular characterization revealed that CA3 genomic DNA consists of seven exons and six introns, spans about 10.5 kb and maps to porcine chromosome 4q11-->q14. Results of expression profiles showed that the expression levels of CA3 increased in skeletal muscles from prenatal 33- to 65-day-old Chinese Tongcheng pigs. These levels subsequently decreased to a steady state in prenatal 90-day-old, postnatal 2-day-old, postnatal 28-day-old, and pregnant 65-day-old pigs. The expression patterns of Chinese Tongcheng pig embryos were different from that of Landrace pig embryos. CA3 was expressed at higher levels in skeletal muscle and liver than in kidney, lung, stomach, intestine, and brain, but was not detected in heart and spleen. Statistical analysis showed the CA3 gene polymorphism was different between Chinese indigenous and introduced commercial western pig breeds, and was associated with intramuscular fat content and percentage of ham of pigs.
A 1320-bp cDNA containing the full coding region of the porcine succinate dehydrogenase complex, subunit D (SDHD) gene was obtained by random sequencing of clones from a Chinese Tongcheng pig 55-day fetal longissimus dorsi muscle cDNA library. Analysis of the SDHD gene across the INRA-University of Minnesota porcine radiation hybrid panel indicated close linkage with microsatellite marker SW2401, located on SSC9p21. The open reading frame of this cDNA covers 480 bp and encodes 159 amino acids. The deduced porcine amino acid sequence showed greater similarity with human and bovine protein sequences than with those from mouse and rat. The BLAST analysis of the porcine SDHD to NCBI identified Unigene Cluster Ssc.2586. Possible single nucleotide polymorphisms (SNP) were identified by alignment of expressed sequence tags in the cluster. The polymerase chain reaction (PCR) single strand conformation polymorphism, sequencing, and PCR restriction fragment length polymorphism were used to confirm and detect a synonymous polymorphic MboI site within the open-reading frame. Allele frequencies of this SNP were investigated in two commercial and five Chinese local pig breeds. These five Chinese breeds had very high frequencies for one allele, whereas frequencies of both alleles were intermediate in Large White and Duroc. An association analysis suggested that different SDHD genotypes have significant differences in loin-muscle area (P < 0.01).
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Implantation is a complicated developmental process. Some hormones and cytokines are involved in the regulation of the implantation by regulating functions of the endometrium and the embryo. Oligosaccharide antigen Lewis Y is specifically expressed on embryo surface and plays an important role in implantation. In this study, effects of Lewis Y on secretion and gene expression of EGF and EGF receptor of the embryo were investigated. Embryos were pre-incubated with Le(Y) specific antibody AH6 for 3 h, then antibody was removed, immuno-dot blots and RT-PCR were used to analyze the secretion of EGF and EGF receptor and their gene expression in the embryo at different times after pre-incubation. The results showed that, when blocked by AH6 antibody for 1.5 h, the secretion and expression of EGF in the embryo were significantly declined, and the alteration was continued to over 6 h; and those of EGF-R were slightly decreased after blocking. The results suggest that Lewis Y oligosaccharide on the surface of embryo may be involved in the regulation of embryo development and implantation, by affecting the expression and secretion of EGF in embryo.
The present study was undertaken to investigate the effects of angiotensin-(1-7)[Ang-(1-7)] on the proliferation and expression of protein kinase C (PKC) and extracellular regulated protein kinases (ERK1/2) of vascular smooth muscle cells (VSMCs) by techniques of Western blot, 3H-thymidine (3H-TdR) and 3H-leucine (3H-Leu) incorporation. Ang II increased VSMC DNA and protein synthesis, and promoted protein expression of PKC-zeta and ERK1/2. In contrast, Ang-(1-7) not only inhibited PKC-zeta and ERK1/2 protein content, but also decreased Ang II mediated VSMC hyperplasia, as compared with the controls (P < 0.01 or P < 0.05). These results suggest that Ang-(1-7) may inhibit VSMC proliferation by influencing protein expression of PKC-zeta and ERK1/2.
The characteristic infrared spectra of the five alpha,alpha'-dioxoketene cyclic dithioacetals such as carbonyl, double bond of carbon and carbon, single bond of carbon and sulfur etc have been studied. It is specially important that the changing rules with the change of chemical structure have been discussed.
AIM: To study the mechanisms of the regulation of the tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor subunit 2B(NR2B) in the gerbil hippocampal synaptosomes following ischemia/reperfusion (I/R). METHODS: Transient (15 min) cerebral ischemia was produced by bilateral carotid artery occlusion procedure. The tyrosine phosphorylation of NR2B was analyzed by immunoprecipitation and immunoblot assay. RESULTS: Transient forebrain ischemia for 15 min caused a marked decrease in the levels of tyrosine phosphorylation of many protein bands including 180 kDa protein. Transient ischemia followed by reperfusion induced rapid (within 15 min of reperfusion), and sustained (for at least 48 h) increase in the tyrosine phosphorylation of many protein bands including 180 kDa protein. Immunoprecipitation and immunoblot confirmed that NR2B is among the phosphorylated 180 kDa protein. Maximal phosphorylation of 180 kDa band corresponding to NR2B (1.8 fold relative to sham-operated controls) was reached at 6 h of reperfusion following 15 min of cerebral ischemia. But the level of protein expression of NR2B did not change. Administration of ketamine (KT), a non-competitive NMDA receptor antagonist, or nifedipine (ND), an L-type voltage gated calcium channel (L-type VGCC) blocker, 20 min before ischemia attenuated stimulation of the tyrosine phosphorylation of NR2B without affecting the level of protein expression of NR2B. Under these conditions, non-NMDA receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) had no effect on the level of tyrosine phosphorylation. Protein tyrosine phosphatase (PTP) inhibitor vanadate and protein tyrosine kinase (PTK) inhibitor genestein resulted in the increase and the decrease of the tyrosine phosphorylation of NR2B, respectively. Src coprecipitated with NR2B protein. CONCLUSION: The increase of the tyrosine phosphorylation of NR2B induced by I/R has relation to NR and L-type VGCC; PTK and PTP participate in the regulation of the tyrosine phosphorylation of NR2B during I/R. Src that associates with NR2B may play an important role in the regulation of the tyrosine phosphorylation of NR2B during I/R.
AIM: To study the changes and mechanisms of protein-tyrosine kinase (PTK) and protein-tyrosine phosphatase (PTP) activities in the hippocamal synaptosome following cerebral ischemia/reperfusion (I/R) in gerbil. METHODS: Transient (15 min) global ischemia was produced by bilateral carotid artery occlusion. Total PTK and PTP activities were measured by [r-32P] incorporation and colorimetric analysis, respectively. Src and proline-rich tyrosine kinase2 (PYK2) activities were measured by immunoprecipitation and [r-32P] incorporation. RESULTS: Total PTK activity increased significantly after I/R, but the PTP activity did not change. The Src activity was much higher than PYK2 activity in sham-operated controls. I/R mainly caused a pronounced increase in Src activity, but not PYK2 activity. The increase in Src activity had no relation to the expression of Src protein. Administration of ketamine (KT) or nifedipine (ND) 20 min before ischemia caused a decrease in total PTK and Src activities, and no change in the PYK2 and PTP activities. CONCLUSION: The increase in PTK activity caused by I/R may be mainly due to the increase in Src activity. This increase in Src activity has no relation to the expression of Src protein. But it is related to the activation of NMDA (N-methyl-D-aspartate) receptor (NR) and L-type voltage-gated calcium channel (L-type VGCC). In other words, the increase in total PTK and Src activities induced by I/R may be mediated via NR and L-type VGCC. The PTP activity did not change during I/R.
Lex [Galbeta1-4(Fucalpha1-3)GlcNAc] and Ley [Fucalpha1-2Galbeta1-4(Fucalpha1-3)GlcNAc] are both stage-specific embryonic antigens. Lex is first detected on the blastomeres of the 8-cell stage embryo, which correlates with the onset of blastomere compaction. Ley is highly expressed on the surface of the blastocyst, which has been shown to be involved in blastocyst attachment in the mouse. In the present study, mouse alpha1,2-FT (also known as FUT1) and alpha1,3-FT (also known as Fuc-TIV), which were responsible for Lex and Ley formation, were examined in preimplantation stage embryos by reverse transcription-PCR and in situ hybridization. alpha1,3-FT mRNA was detected in all embryos of preimplantation stage, while alpha1,2-FT mRNA emerged in the later stage embryos from 8-cell to 16-cell to the blastocyst. These results indicated the expression of Ley was regulated by alpha1,2-FT. In situ hybridization showed that these two enzyme mRNAs were detected only in morula and blastocyst stage embryos. The alpha1,2-FT and alpha1, 3-FT mRNAs were located in both the inner cell mass and the trophoblast cells. 2-Cell and 4-cell embryos were isolated from the oviduct and cultured in vitro to the 8-cell, morula and blastocyst stage. The expression of alpha1,2-FT and alpha1,3-FT were observed in these embryos developed in vitro; immunohistochemical analysis also showed that Ley expression was positive. These results suggested the stage-specific expression of Ley on the embryos was synthesized by endogenous alpha1,2-FT and alpha1,3-FT rather than transfer from other sources. In addition, the expression of alpha1, 2-FT was differentially regulated and the uterine factor was not prerequisite of the expression of Ley.