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Selected AbstractsCloning of FSH-,, LH-, and glycoprotein hormone , subunits in pejerrey Odontesthes bonariensis (Valenciennes): expression profile and relationship with GnRH expression and plasma sex steroid levels in male fishJOURNAL OF FISH BIOLOGY, Issue 6 2007L. A. Miranda Three cDNAs encoding pejerrey Odontesthes bonariensis follicle stimulating hormone-, (FSH-,), luteinizing hormone-, (LH-,) and glycoprotein-, (GPH-,) subunits were cloned and characterized. Gene expression of these subunits was analysed by real-time polymerase chain reaction (PCR) and compared with the brain gene expression of endogenous gonadotropin-releasing hormones (GnRHs): Pacific salmon GnRH (GnRH-III), pejerrey GnRH (GnRH-I) and chicken GnRH-II (GnRH-II) and plasma sex steroid levels in adult males. The nucleotide sequences of the FSH-,, LH-, and GPH-, subunits are 466, 558 and 677 base pairs long, encoding for mature peptides of 102, 118 and 98 amino acids respectively. Maturing males had high expression of FSH-, and GPH-, subunits, and intermediate levels of LH-, when compared with running ripe and spent stages. These animals had the lowest plasma testosterone (T) and 11-ketosterone (11-KT) values as well as low expression of sGnRH, cGnRH-II and pjGnRH. Running ripe males had the lowest expression of FSH-, and the highest expression of LH-, and GPH-, subunits, and of the three GnRH genes. At this stage, the highest values of T and 11-KT were observed. Spent males showed low expression of the three gonadotropin (GtH) subunits, sGnRH, pjGnRH and low levels of T. At this stage, 11-KT levels and cGnRH-II expression showed a tendency to decrease but the values were not statistically significant (P < 0·05) to running ripe stage. The present results would suggest that T and 11-KT modulate the expression of the FSH subunits. The expression of the anterior brain GnRH variants, sGnRH and pjGnRH is correlated with LH-, expression and reinforce the importance of the forebrain GnRH variants on the regulation of pituitary function. [source] Cloning and expression analysis of a cDNA encoding lipoprotein lipase from the liver of adult grass carp (Ctenopharyngodon idella)AQUACULTURE RESEARCH, Issue 16 2009Han-Liang Cheng Abstract A full-length cDNA coding lipoprotein lipase (LPL) was cloned from the liver of adult grass carp (Ctenopharyngodon idella) using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends approaches. The cDNA obtained was 2414 bp long with a 1524 bp open reading frame encoding 507 amino acids, including a putative signal peptide 21 amino acids long. The LPL protein has a calculated molecular weight of 57.77 kDa and an isolectric point of 8.132. The main domains of LPL, such as catalytic site, disulphide bridge, N-linked glycosylation site, heparin-binding domain, lipid-binding site and site of dimer formation, are basically conserved between the grass carp and other vertebrates. The tissue distribution of LPL mRNA in the liver, head kidney, mesenteric adipose tissue, heart and white muscle of adult grass carp was analysed using the semi-quantitative RT-PCR method using ,-actin gene as an internal control; the result showed that the expressions of LPL mRNA were detected in all examined tissues of adult grass carp. The expression levels of LPL in the mesenteric adipose tissue were the highest among these tissues, followed by the liver and head kidney and the lowest expression was found in the heart and white muscle. [source] Differential expression of stromal cell,derived factor 1 and its receptor CXCR4 in the skin and endothelial cells of systemic sclerosis patients: Pathogenetic implicationsARTHRITIS & RHEUMATISM, Issue 9 2006Paola Cipriani Objective Systemic sclerosis (SSc) is characterized by early endothelial damage evolving to vascular desertification. Stromal cell,derived factor 1 (SDF-1) and its receptor CXCR4 regulate specific steps in new vessel formation. We undertook this study to determine whether an alteration of the SDF-1/CXCR4 axis might be involved in the pathogenetic mechanisms following ischemic damage during SSc. Methods We enrolled 36 SSc patients and 15 controls. Skin biopsy samples were obtained from each subject, and the expression of SDF-1 and CXCR4 was assessed by immunohistochemistry, reverse transcription,polymerase chain reaction (RT-PCR), and Western blot analyses. Furthermore, isolated microvascular endothelial cells (MVECs) from 4 patients with diffuse cutaneous SSc (dcSSc) and 3 controls were analyzed for SDF-1 and CXCR4 by confocal laser scanning microscopy, RT-PCR, and Western blotting. Results SDF-1 and CXCR4 were up-regulated in the skin of patients with early (edematous) SSc, both in the diffuse and limited cutaneous forms, and progressively decreased, with the lowest expression in the latest phases of both SSc subsets. MVECs from patients with dcSSc expressed significantly higher amounts of both isoforms of SDF-1 in the early stage of disease, with a progressive reduction of SDF-1 and CXCR4 in later stages. On the surface of cultured MVECs from patients with dcSSc, SDF-1 and CXCR4 colocalized in polarized areas, suggesting that they are activated in vivo and that they are under strict genetic control to retain capping function. Conclusion Due to its transient expression, SDF-1 could be considered a future therapeutic target to induce new vessel formation in SSc. [source] Substance P and CGRP expression in dental pulps with irreversible pulpitisAUSTRALIAN ENDODONTIC JOURNAL, Issue 2 2010Mandana Sattari dds Abstract The purpose of this study was to compare substance P (SP) and calcitonin gene-related peptide (CGRP) expression in pulp tissue with clinically diagnosed symptomatic and asymptomatic irreversible pulpitis. Healthy pulps acted as controls. Five normal pulps and 40 with irreversible pulpitis (20 symptomatic and 20 asymptomatic) were obtained from 45 different patients. SP and CGRP expression was determined by competition binding assays using enzyme immunoassay. anova and Mann,Whitney tests were used to ascertain if there were statistically significant differences between the groups. The results showed that neuropeptides were found in all pulp samples. The highest and the lowest expressions for SP and CGRP were found in symptomatic irreversible pulpitis and healthy pulps groups, respectively. The differences between healthy pulps and the groups of pulps having irreversible pulpitis were significant (P < 0.001). Although Mann,Whitney's post-hoc tests showed statistically significant differences in CGRP expression between two pulpitis groups (P < 0.05), differences in SP expression between symptomatic and asymptomatic irreversible pulpitis groups were not significant. This study demonstrated that the expression of CGRP and SP is significantly higher in pulps with irreversible pulpitis compared with healthy pulps. [source] |