1.Sialoglycoproteins and penultimate sugar expression pattern in developing murine olfactory and respiratory mucosa.
Joo Heon YOON ; Kyung Su KIM ; Sung Shik KIM ; Jeung Gweon LEE
Yonsei Medical Journal 1998;39(1):20-26
Sialic acid residues are constant constituents of the glycoproteins of the airways in all species. Sialoglycoproteins are the main acidic glycoprotein and their functions are to mediate cell adherence, to control the viscoelasticity of mucus and to serve as receptor sites for the binding of exogenous macromolecules. The purpose of this study was to investigate the differences in the distribution of sialoglycoproteins as a terminal sugar and in the composition of the penultimate sugar according to aging in the murine nasal respiratory and olfactory mucosa. Nasal cavities of mice (BALB/c) were fixed by intracardiac perfusion with 2.0% glutaraldehyde and embedded in Epon 812. First, the serial sections were stained with Maackia amurensis agglutinin (MAA) and Sambucus nigra agglutinin (SNA). Then, the adjacent sections were stained with DBA and PNA before and after neuraminidase digestion in all experimental groups. Apical cell surfaces of olfactory mucosa and cilia on a few ciliated cells in the mucosa of the septum and nasal floor were labelled with MAA, but cell surfaces of respiratory mucosa, Bowman's glands and goblet cells were not labelled with MAA, irrespective of aging. Apical cell surfaces of both olfactory and respiratory mucosa and Bowman's glands were stained with SNA, however, goblet cells were not labelled with SNA. After neuraminidase digestion to remove terminal sialic acid residues of sialoglycoproteins, only cell surfaces of respiratory mucosa were labelled with PNA, but goblet cells, cell surfaces of olfactory mucosa and Bowman's glands were not labelled with PNA. Cell surfaces and Bowman's glands of olfactory mucosa were labelled with DBA after neuraminidase digestion, but cell surfaces of respiratory mucosa and goblet cells were not labelled with DBA. Our results indicate that there were different carbohydrate structures of sialoglycoconjugates in olfactory and respiratory mucosa, and it was not influenced by aging.
Aging/metabolism*
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Animal
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Carbohydrates/analysis*
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Mice
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Mice, Inbred BALB C
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Nasal Mucosa/chemistry*
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Olfactory Mucosa/chemistry*
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Sialoglycoproteins/analysis*
2.Hypoxic condition promotes olfactory mucosa mesenchymal stem cells to differentiate into neurons and underlying mechanisms.
Yi ZHUO ; Ting YUAN ; Da DUAN ; Lei WANG ; Lite GE ; Pei WU ; Hao WANG ; Ming LU
Journal of Central South University(Medical Sciences) 2016;41(12):1252-1259
To explore whether hypoxic condition could promote the olfactory mucosa mesenchymal stem cells (OM-MSCs) to differentiate into neurons with the olfactory ensheathing cells (OECs) supernatant and the potential mechanisms.
Methods: The OM-MSCs and OECs were isolated and cultured, and they were identified by flow cytometry and immunofluorescence. The OM-MSCs were divided into three groups: a 3%O2+ HIF-1α inhibitors (lificiguat: YC-1) + OECs supernatant group (Group A) , a 3%O2 + OECs supernatant group (Group B) and a 21%O2 + OECs supernatant group (Control group). The neurons, which were differentiated from OM-MSCs, were assessed by immunofluorescence test. The mRNA and protein expression of hypoxia-inducible factor-1α (HIF-1α), βIII-tubulin and glial fibrillary acidic portein (GFAP) were detected by quantitative polymerase chain reaction (Q-PCR) and Western blot. The potassium channels were analyzed by patch clamp.
Results: The neurons differentiated from OM-MSCs expressed the most amount of βIII-tubulin, and the result of Q-PCR showed that HIF-1α expression in the Group B was significantly higher than that in the other groups (all P<0.05). Western blot result showed that the βIII-tubulin protein expression was significantly higher and GFAP protein expression was obviously decreased in the Group B (both P<0.05). The patch clamp test confirmed that the potassium channels in the neurons were activated.
Conclusion: Hypoxic condition can significantly increase the neuronal differentiation of OM-MSCs by the OECs supernatant and decrease the production of neuroglia cells, which is associated with the activation of HIF-1 signal pathway.
Blotting, Western
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Cell Differentiation
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physiology
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Cells, Cultured
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Culture Media, Conditioned
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chemistry
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pharmacology
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Flow Cytometry
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Glial Fibrillary Acidic Protein
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metabolism
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Hypoxia
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physiopathology
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Hypoxia-Inducible Factor 1, alpha Subunit
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metabolism
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Indazoles
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pharmacology
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Mesenchymal Stem Cells
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physiology
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Neurogenesis
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physiology
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Neuroglia
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metabolism
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physiology
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Neurons
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physiology
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Olfactory Mucosa
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Potassium Channels
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Signal Transduction
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Tubulin
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metabolism