Assuntos
Caveolina 1/metabolismo , Glicoproteínas/química , Glicoesfingolipídeos , Microdomínios da Membrana/metabolismo , Neoplasias , Caveolina 1/genética , Adesão Celular , Comunicação Celular , Movimento Celular , Expressão Gênica , Glicoproteínas/genética , Glicoproteínas/metabolismo , Glicoesfingolipídeos/química , Glicoesfingolipídeos/metabolismo , Glicosilação , Humanos , Microdomínios da Membrana/patologia , Invasividade Neoplásica , Neoplasias/química , Neoplasias/metabolismo , Neoplasias/patologia , Transdução de Sinais , Microambiente TumoralRESUMO
There is a clear need to develop novel tools to help improve our understanding of stem cell biology, and potentially also the utility of stem cells in regenerative medicine. We report the cloning, functional, and bioinformatic characterization of a novel stem cell marker in the zebrafish retina, drCol 15a1b. The expression pattern of drCol 15a1b is restricted to stem cell niches located in the central nervous system, whereas other collagen XVs are associated with muscle and endothelial tissues. Knocking down drCol 15a1b expression causes smaller eyes, ear defects, and brain edema. Microscopic analysis reveals enhanced proliferation in the morphant eye, with many mitotic nuclei located in the central retina, together with a delayed differentiation of the mature retinal cell types. Besides, several markers known to be expressed in the ciliary marginal zone display broader expression areas in morpholino-injected embryos, suggesting an anomalous diffusion of signaling effectors from the sonic hedgehog and notch pathways. These results indicate that drCol 15a1b is a novel stem cell marker in the central nervous system that has a key role in homing stem cells into specialized niches in the adult organism. Moreover, mutations in the hCol 18a1 gene are responsible for the Knobloch syndrome, which affects brain and retinal structures, suggesting that drCol 15a1b may function similarly to mammalian Col 18a1. Thus, our results shed new light on the signaling pathways that underlie the maintenance of stem cells in the adult organism while helping us to understand the role of extracellular matrix proteins in modulating the signals that determine stem cell differentiation, cell cycle exit and apoptosis.
Assuntos
Retina/metabolismo , Nicho de Células-Tronco/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Animais , Colágeno/classificação , Colágeno/genética , Colágeno/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Filogenia , Reação em Cadeia da Polimerase , Retina/embriologia , Peixe-Zebra , Proteínas de Peixe-Zebra/classificação , Proteínas de Peixe-Zebra/genéticaRESUMO
In this paper, we describe the effects of the expression of GM3 synthase at high levels in human ovarian carcinoma cells. Overexpression of GM3 synthase in A2780 cells consistently resulted in elevated ganglioside (GM3, GM2 and GD1a) levels. GM3 synthase overexpressing cells had a growth rate similar to wild-type cells, but showed a strongly reduced in vitro cell motility accompanied by reduced levels of the epithelial-mesenchymal transition marker alpha smooth muscle actin. A similar reduction in cell motility was observed upon treatment with exogenous GM3, GM2, and GM1, but not with GD1a. A photolabeling experiment using radioactive and photoactivable GM3 highlighted several proteins directly interacting with GM3. Among those, caveolin-1 was identified as a GM3-interacting protein in GM3 synthase overexpressing cells. Remarkably, caveolin-1 was markedly upregulated in GM3 synthase overexpressing cells. In addition, the motility of low GM3 synthase expressing cells was also reduced in the presence of a Src kinase inhibitor; on the other hand, higher levels of the inactive form of c-Src were detected in GM3 synthase overexpressing cells, associated with a ganglioside- and caveolin-rich detergent insoluble fraction.