ABSTRACT
Objective: To uncover the time-dependent expression pattern of ptk2b gene and ptk2b-encoded protein, protein tyrosine kinase 2 beta(PTK2B), in the brain tissues of transgenic animal models of Alzheimer's disease (AD) and its relationship with the levels of Aβ1-42, phosphorylation of Tau (p-Tau) and low density lipoprotein receptor-related protein-1(LRP-1) in blood and brain tissues. Methods: In this study, 5-, 10- and 15-month-old APPswe/PS1dE9 double-transgenic mice harboring the genotype of AD confirmed by the gene test were divided into the 5-, 10- and 15-month-old experiment groups, and simultaneously, age-matched C57BL/6J mice were placed into the corresponding control groups, with 8 mice in each group. All mice were subjected to the Morris Water Maze for test of cognitive and behavioral ability. Expression profiles of PTK2B, Aβ1-42, p-Tau/Tau and LRP-1 in the hippocampus or blood of mice were quantified by using the immunohistochemistry staining, Western blot or enzyme-linked immunosorbent assay (ELISA), while the mRNA expression of ptk2b in the hippocampus was quantified by using the real-time quantitative polymerase chain reaction (qRT-PCR). Results: Results of experiment groups demonstrated that as mice aged, the expression levels of PTK2B, ptk2b mRNA, Aβ1-42 and p-Tau/Tau in the hippocampus were increased, and the expression of LRP-1 was decreased gradually. While in the blood, the level of Aβ1-42 was decreased, and the cognitive and behavioral ability was decreased in an age-dependent manner (all P＜ 0.05). However, comparisons among the control groups, only the age-dependent downregulation of LRP-1 were observed in hippocampus(P＜0.05), but other indicators had no significant differences (P＞0.05). Conclusion: In the hippocampus of APP/PS1 double-transgenic mice, the expressions of PTK2B, Aβ1-42 and p-Tau/Tau are upregulated, LRP-1 is downregulated, while cognitive and behavioral ability is decreased, and such changes are presented in a time-dependent manner.
Subject(s)
Animals , Mice , Alzheimer Disease/metabolism , Amyloid beta-Peptides , Amyloid beta-Protein Precursor/genetics , Focal Adhesion Kinase 2/metabolism , Hippocampus/metabolism , Low Density Lipoprotein Receptor-Related Protein-1 , Maze Learning , Mice, Inbred C57BL , Mice, Transgenic , RNA, MessengerABSTRACT
OBJECTIVE: The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes. METHODS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors. RESULTS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells. CONCLUSIONS: Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors. .
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Bone Marrow Cells/metabolism , /metabolism , Myelodysplastic Syndromes/metabolism , /analysis , /analysis , Focal Adhesion Protein-Tyrosine Kinases/analysis , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Myelodysplastic Syndromes/genetics , Polymerase Chain Reaction , Prognosis , /metabolism , Risk Factors , Statistics, Nonparametric , src Homology Domains/physiologyABSTRACT
Objective To investigate the changes of gene expression in the cardiac outflow tract (OFT) in Cx43 knockout (Cx43-/-) mouse embryos, and to elucidate the genes involved in the myocardialization of proximal cardiae outflow tract septum. Methods The cDNA was retrotranscripted from RNA, which extracted from OFT tissues of both Cx43-/- and Cx43 wild type (Cx43+/+) mouse embryos on embryonic day (ED) 14.5. The biotin-labeled cRNA derived from the transcription of cDNA was fragmented as probes. The probes were hybridized with Affymetrix Mouse Genome 430 2.0 Array. Gene Array Scanner was used to screen the signals of hybridization, and the expression of genes was detected. Results Among the differentially expressed genes, there were 143 upregulated and 235 downregulated in Cx43 knockout OFT tissue compared with those in Cx43+/+ heart. Functions of proteins encoded by the altered genes encompassed all functional categories, with the largest percentage in genes involved in signaling pathways such as regulation of transcription, cell cycle, etc. Among the differentially expressed genes in the Cx43-/- heart, some were related with TGFβ/BMP signaling pathway, and Ssr1, Ptk2 and Bmp6 were related with conotruncal defects. These genes were verified by Real-time PCR, and the result was consistent with that of microarray (P<0.05).Conclusions Scaned by Gene Array and verified by Real-time PCR, genes related with TGFβ/BMP signaling pathway, and Ssr1, Ptk2 and Bmp6 were differentially expressed in ED 14.5 Cx43-/- OFT tissue, which may be involved in the myocardialization of proximal cardiac outflow tract septum.