ABSTRACT
A novel 3,4,4a-trihydroxanthene-fused pyrrole 2 was synthesized by the reaction of 2,3,4,4a-tetrahydro-1H-xanthen-1-one with 3-phenyl-2H-azirine in the presence of LDA. Utilizing this pyrrole 2, a NIR BODIPY 1 (λ(abs) = 732 nm, λ(em) = 747 nm) has been prepared. The new BODIPY 1 was stable, non-cytotoxic, and suited to labeling living cells for imaging assay in the NIR region.
Subject(s)
Boron Compounds/chemistry , Fluorescent Dyes/chemistry , Infrared Rays , Xanthenes/chemistry , Boron Compounds/chemical synthesis , Boron Compounds/metabolism , Boron Compounds/toxicity , Fluorescent Dyes/chemical synthesis , Fluorescent Dyes/metabolism , Fluorescent Dyes/toxicity , Hep G2 Cells , Humans , Pyrroles/chemistryABSTRACT
BACKGROUND: Neuron-derived neurotrophic factor (NDNF) is evolutionarily well conserved, being present in invertebrate animals such as the nematode, Caenorhabditis elegans, as well as in the fruit fly, Drosophila melanogaster. Multiple cysteines are conserved between species and secondary structure prediction shows that NDNF is mainly composed of beta-strands. In this study, we aimed to investigate the function of NDNF. RESULTS: NDNF is a glycosylated, disulfide-bonded secretory protein that contains a fibronectin type III domain. NDNF promoted migration and growth and elicited neurite outgrowth of mouse hippocampal neurons in culture. NDNF also protected cultured hippocampal neurons against excitotoxicity and amyloid beta-peptide toxicity. Western blotting showed that NDNF was exclusively expressed in the brain and spinal cord. Immunostaining indicated that NDNF was expressed by neurons and not by astrocytes. Cajal-Retzius cells, cortex neurons, hippocampus neurons, olfactory mitral cells, cerebellar purkinje cells, cerebellar granular cells and spinal neurons were found to be NDNF-positive. NDNF expression was observed in the neurons during development. CONCLUSIONS: The results of this study indicated that NDNF is a novel neurotrophic factor derived from neurons that may be useful in the treatment of neuronal degeneration diseases and nerve injuries.
Subject(s)
Brain/embryology , Brain/metabolism , Cell Differentiation/genetics , Nerve Growth Factors/biosynthesis , Neurons/metabolism , Amino Acid Sequence , Animals , Anura , Base Sequence , Brain/growth & development , Cattle , Cell Differentiation/physiology , Cells, Cultured , Chickens , Cytoprotection/genetics , Cytoprotection/physiology , Drosophila , Gene Expression Regulation, Developmental/genetics , HEK293 Cells , Humans , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Nematoda , Nerve Growth Factors/chemistry , Nerve Growth Factors/genetics , Neurogenesis/physiology , Neurons/cytology , RatsABSTRACT
OBJECTIVE: To investigate the relations between neuroapoptosis and the onset and development of Alzheimer's disease (AD), especially the role of NF-kappaB in the regulation of neuroapoptosis. METHODS: Caspase-3 and NF-kappaB (p50) expressions in the CA3 region of the hippocampus in APPswe Tg2576 transgenic mice were studied from postnatal day 0-180, using Nissl staining, immunohistochemistry and RT-PCR methods. RESULTS: Both neuronal apoptosis and NF-kappaB activity decreased gradually with the increase of age in wild type and Tg2576 mice. However, the number of caspase-3-positive or NF-kappaB-positive pyramidal cells in Tg2576 mice was greater than that in age-matched wild type mice, with significant differences after postnatal day 14 (P < 0.01 or P < 0.05). Linear regression analyses of caspase-3 and NF-kappaB expression demonstrated a correlation between neuroapoptosis and activity of NF-kappaB. CONCLUSION: The process of neuroapoptosis is consistent with the onset and development of AD. Furthermore, the observed correlation between neuroapoptosis and NF-kappaB activity suggests a role of NF-kappaB in hippocampal neuroapoptosis.
Subject(s)
Apoptosis/physiology , CA3 Region, Hippocampal/growth & development , CA3 Region, Hippocampal/metabolism , Caspase 3/metabolism , NF-kappa B/metabolism , Pyramidal Cells/metabolism , Aging/metabolism , Aging/pathology , Alzheimer Disease , Animals , Animals, Newborn , CA3 Region, Hippocampal/pathology , Cell Count , Disease Models, Animal , Mice , Mice, Inbred C57BL , Mice, Transgenic , Neurons/metabolism , Neurons/pathology , Pyramidal Cells/pathology , RNA, Messenger/metabolismABSTRACT
ATP-binding cassette transporter A1 (ABCA1) modulates plasma levels of high density lipoprotein (HDL), a cardiovascular protecting factor. Tree shrew was considered to be an animal protected from atherosclerosis characterized by high proportion of HDL in plasma. The cDNA clones and expression of tree shrew ABCA1 was identified using SMART-RACE and Real-Time PCR techniques respectively. The nucleotide sequence of tree shrew ABCA1 covered 7,762 bp, including a 6,786 bp coding region which encoded a 2,261 amino acids protein with the high identity to human ABCA1 (95%). Tree shrew ABCA1 was expressed in various tissues, the highest in lung, followed by liver, kidney, spleen and cardiac muscle in turn from high to medium expression levels. This pattern was partially different from that of human ABCA1 which was low in kidney and cardiac muscle. This work could shed new light on its role of ABCA1 in the distinctive HDL metabolism in tree shrew.
