Γονιδιακή κλωνοποίηση μορίου κυτταρικής συνάφειας του νευρικού συστήματος

The molecular cloning, characterization and identification of biological function of the cell adhesion molecules in the nervous system of vertebrates and invertebrates will be the guide to our understanding of the molecular mechanisms involved in the development of the nervous system (cell migration, differentiation, cell aggregation, growth and fasciculation of axons, synaptic connections). In this work, LN-10 cell adhesion molecule (chick) has been cloned with the use of heterologous cDNA probe (rat). The LN-10 molecule is expressed specifically in the chick nervous system (cerebellum, optic lobus, hemispheres) and is absent from non-neuronal tissues (liver, blood, intestine). The expression of LN-10 molecule increases gradually until post hatching day 2 and a decrease follows. LN-10 mRNA is expressed in granule cells of chick cerebellum and comparison with other molecules shows homology to Elastin precursors, chick N-Cadherin and Drosophila Notch protein. The study of the LN-10 molecule was carried out with the techniques: synthesis and screening cDNA bank, Southern, Northern and Western blot, in situ RNA-RNA hybridization and DNA sequencing. The results of the above studies are as follows: 1. Western blot analysis revealed NILE protein molecules in chick cerebellum. Their size differs to that reported for another chick CAM, namely NgCAM. 2. NILE-related P12-13 cDNA clone from PC12 cells exhibits homology with chick genomic DNA fragments, in Southern blot analysis. 3. Three cDNA clones have been detected from λgt11 cDNA library from 2-days old chick cerebellum, with the use of P12-13 cDNA clone as heterologous probe (LN-9 0.5kb, LN-11 1.8kb, LN-10 2.0kb). 4. The LN-10 clone derives most probably from a single gene, which contains at least one intron as shown by restriction enzyme analysis. 5. The largest messanger, which is detected from LN-10 probe in cerebellum is approx. 2.0kb long. 6. The LN-10 clone is absent in non-neuronal tissues while are detected in optic lobus and hemispheres more messangers (2.0, 4.2, 5.2 and 6.8kb), which possibly result from alternative splicing of a single gene transcription product. 7. The RNA detected by the LN-10 clone is present in chick cerebellum from E14 embryonic day, increases the days around hatching and is almost not expressed at post-hatching day 17. 8. The nucleotide sequence of LN-10 clone was determined (1941 nucleotides). It contains an ORF 1629 nt and probably it synthesizes a polypeptide sequence of 543 amino acids. 9. Genomic clones of LN-10 molecule were isolated from λEMBL₄ chick genomic library.

Grantor dc:publisher

University of Patras

Year dc:date

1990

Authors dc:creator

• Zarkadis, Ioannis
• Ζαρκάδης, Ιωάννης

• Αναπτυξιακή ρύθμιση
• Γονιδιακή κλωνοποίηση
• Μόριο κυτταρικής συνάφειας
• Νευρικό σύστημα
• Cell adhesion molecule
• Developmental regulated
• Molecular cloning
• Nervous specific
• Ιατρική και Επιστήμες Υγείας
• Επιστήμες Υγείας
• Medical and Health Sciences
• Health Sciences

Language dc:language

gre

Identifier

10.12681/eadd/1395

OAI identifier oai:identifier

oai:10442/1395