Cell adhesion molecules belonging to the immunoglobulin superfamily play apivotal role in different stages of neuronal development and also partecipatein the regulation of many aspects of synaptic stabilization. The mouse axonalglycoprotein F3/Contactin is an adhesion molecule that is involved in axonaloutgrowth and guidance and mediates interactions between neural cells. Inprevious studies, by applying antibodies against the mouse F3/contactinprotein, we have found that an F3/Contactin related molecule (HFRM) isexpressed in the nervous system of the snail Helix pomatia. This moleculeappears to be important for cell adhesion and neurite outgrowth of Helixneurons in culture and it is involved in the targetdependentregulation ofneurotransmitter release from presynaptic terminals.In these experiments, by screening with anti F3/Contactin antiserum a cDNAexpression library prepared from Helix nervous system, we isolated the 9.1lambda clone, which gave the strongest signal on the immunoscreeningprocedure. We obtained a full length cDNA(3522 bp) by both 5'and 3'RACEPCRapproaches using oligonucleotides derived from the cloned sequence.Translation of this cDNArevealed an open reading frame encoding a 718aminoacid sequence including a signal peptide, 5 fibronectin type III repeatsand 9 potential Nglycosilationsites indicating that the molecule couldcorrespond to a secreted protein. Finally, Blast analysis showed more than30% of homology with the mouse F3/contactin molecule in the fibronectintype III regions, demonstrating a structural correlation between the newlyfound protein of Helix pomatia and known cell adhesion molecules involvedin the nervous system development.
Cloning of an F3/Contactinrelated molecule in the mollusc Helix pomatia nervous system
Milanese, Chiara;
2004-01-01
Abstract
Cell adhesion molecules belonging to the immunoglobulin superfamily play apivotal role in different stages of neuronal development and also partecipatein the regulation of many aspects of synaptic stabilization. The mouse axonalglycoprotein F3/Contactin is an adhesion molecule that is involved in axonaloutgrowth and guidance and mediates interactions between neural cells. Inprevious studies, by applying antibodies against the mouse F3/contactinprotein, we have found that an F3/Contactin related molecule (HFRM) isexpressed in the nervous system of the snail Helix pomatia. This moleculeappears to be important for cell adhesion and neurite outgrowth of Helixneurons in culture and it is involved in the targetdependentregulation ofneurotransmitter release from presynaptic terminals.In these experiments, by screening with anti F3/Contactin antiserum a cDNAexpression library prepared from Helix nervous system, we isolated the 9.1lambda clone, which gave the strongest signal on the immunoscreeningprocedure. We obtained a full length cDNA(3522 bp) by both 5'and 3'RACEPCRapproaches using oligonucleotides derived from the cloned sequence.Translation of this cDNArevealed an open reading frame encoding a 718aminoacid sequence including a signal peptide, 5 fibronectin type III repeatsand 9 potential Nglycosilationsites indicating that the molecule couldcorrespond to a secreted protein. Finally, Blast analysis showed more than30% of homology with the mouse F3/contactin molecule in the fibronectintype III regions, demonstrating a structural correlation between the newlyfound protein of Helix pomatia and known cell adhesion molecules involvedin the nervous system development.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
