The lipophosphoglycan (LPG) of Leishmania major has a major role in the attachment to Phlebotomus papatasi midgut. Here, we investigated the comparative structural features of LPG of L. turanica, another species transmitted by P. papatasi. The mAb WIC 79.3, specific for terminal Gal(β1,3) side-chains, strongly reacted with L. turanica LPG. In contrast, L. turanica LPG was not recognized by arabinose-specific mAb 3F12. In conclusion, LPGs from L. major and L. turanica are similar, with the latter being less arabinosylated than L. major's. The high galactose content in L. turanica LPG is consistent with its predicted recognition by P. papatasi lectin PpGalec.
- MeSH
- druhová specificita MeSH
- glykosfingolipidy chemie genetika metabolismus MeSH
- hmyz - vektory parazitologie MeSH
- Leishmania genetika metabolismus MeSH
- Phlebotomus parazitologie MeSH
- regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Binding of promastigotes to the sand fly midgut epithelium is regarded as an essential part of the Leishmania life cycle in the vector. Among Leishmania surface molecules putatively involved in attachment to the sand fly midgut, two GPI-anchored molecules are the most prominent: lipophosphoglycan (LPG) and promastigote surface protease gp63. In this work, we examined midgut attachment of Leishmania lines mutated in GPI-anchored molecules and compared results from 2 different techniques: in vivo development in sand flies and in vitro competitive binding assays using fluorescently labelled parasites. In combination with previous studies, our data provide additional support for (1) an LPG-independent parasite-binding mechanism of Leishmania major within the midgut of the permissive vector Phlebotomus perniciosus, and provide strong support for (2) the crucial role of L. major LPG in specific vector Phlebotomus papatasi, and (3) a role for Leishmania amazonensis gp63 in Lutzomyia longipalpis midgut binding. Moreover, our results suggest a critical role for GPI-anchored proteins and gp63 in Leishmania mexicana attachment to L. longipalpis midguts, as the wild type (WT) line accounted for over 99% of bound parasites.
- MeSH
- galaktosyltransferasy genetika metabolismus MeSH
- glykokonjugáty genetika metabolismus MeSH
- glykosfingolipidy genetika metabolismus MeSH
- hmyz - vektory parazitologie MeSH
- kompetitivní vazba MeSH
- Leishmania fyziologie MeSH
- lidé MeSH
- metaloendopeptidasy genetika metabolismus MeSH
- mutace MeSH
- Phlebotomus parazitologie MeSH
- protozoální proteiny genetika metabolismus MeSH
- Psychodidae parazitologie MeSH
- stadia vývoje MeSH
- trávicí systém parazitologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- Fabryho nemoc etiologie MeSH
- Gaucherova nemoc MeSH
- glykogenóza typu II MeSH
- glykogenóza typu IIb MeSH
- glykosfingolipidy genetika škodlivé účinky MeSH
- hemochromatóza komplikace MeSH
- karnitin nedostatek MeSH
- lidé MeSH
- metabolické nemoci * MeSH
- mitochondriální nemoci * komplikace MeSH
- mukopolysacharidózy komplikace MeSH
- nemoci srdce * komplikace MeSH
- přetížení železem komplikace MeSH
- Check Tag
- lidé MeSH
Various types of feeder cells have been adopted for the culture of human embryonic stem cells (hESCs) to improve their attachment and provide them with stemness-supporting factors. However, feeder cells differ in their capacity to support the growth of undifferentiated hESCs. Here, we compared the expression and secretion of four well-established regulators of hESC pluripotency and/or differentiation among five lines of human foreskin fibroblasts and primary mouse embryonic fibroblasts throughout a standard hESC culture procedure. We found that human and mouse feeder cells secreted comparable levels of TGF beta 1. However, mouse feeder cells secreted larger quantities of activin A than human feeder cells. Conversely, FGF-2, which was produced by human feeder cells, could not be detected in culture media from mouse feeder cells. The quantity of BMP-4 was at about the level of detectability in media from all feeder cell types, although BMP-4 dimers were present in all feeder cells. Production of TGF beta 1, activin A, and FGF-2 varied considerably among the human-derived feeder cell lines. Low- and high-producing human feeder cells as well as mouse feeder cells were evaluated for their ability to support the undifferentiated growth of hESCs. We found that a significantly lower proportion of hESCs maintained on human feeder cell types expressed SSEA3, an undifferentiated cell marker. Moreover, SSEA3 expression and thus the pluripotent hESC compartment could be partially rescued by addition of activin A. Cumulatively, these results suggest that the ability of a feeder layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production.
- MeSH
- aktiviny biosyntéza genetika MeSH
- antigeny sacharidové asociované s nádorem genetika MeSH
- biologické markery metabolismus MeSH
- buněčná diferenciace MeSH
- DNA primery genetika MeSH
- druhová specificita MeSH
- embryonální antigeny specifické pro určité stadium vývoje MeSH
- embryonální kmenové buňky cytologie metabolismus metabolismus MeSH
- exprese genu MeSH
- fibroblastový růstový faktor 2 biosyntéza genetika MeSH
- financování organizované MeSH
- glykosfingolipidy genetika MeSH
- kokultivační techniky metody MeSH
- kostní morfogenetické proteiny analýza genetika MeSH
- kostní morfogenetický protein 4 MeSH
- kultivační média speciální analýza MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- myši MeSH
- pluripotentní kmenové buňky cytologie metabolismus MeSH
- proliferace buněk MeSH
- sekvence nukleotidů MeSH
- transformující růstový faktor beta1 biosyntéza genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
