In situ Hybridization of Chemotactically Bioactive Molecules on Whole Mounted Chick Embryo.

TOLOSA EJ, JAURENA MB, ZANIN JP, BATTIATO NL, ROVASIO RA

JOURNAL OF HISTOTECHNOLOGY

NATL SOC HISTOTECHNOLOGY

Año: 2012

0147-8885

Chemotaxis modulates cell distribution in response to concentration gradients of molecules released by target fields. Objective.  To show that chemotactic responses of in vitro neural crest cells (NCCs) induced by the chemokine Stromal Cell-Derived Factor-1 (SDF-1), the growth factor Neurotrophin-3 (NT-3) and the morphogen Sonic Hedgehog (Shh), are associated with a topological distribution of these molecules on cultured chick embryo. Methods.  Whole mount in situ hybridization to show the expression of mRNA transcripts of SDF-1, NT-3 and Shh, protein immunolocalization and immunolabeling of NCCs, in early embryo development stages. Results.  Spatiotemporal expression of bioactive molecules was shown at early embryo cephalic sites, the target fields of NCCs, supporting the in vivo chemotactic behavior of this cell population. This indicates that directional migration of mesencephalic NCCs is modulated by guiding molecules segregated from the optic vesicle region, where NCCs colonize and differentiate into neurones and glia of the ciliary ganglia. Discussion.  The capacity to visualize the in situ expression of molecules involved in the chemotactic response of NCCs to concentration gradients established from target regions is relevant intrinsically and for the knowledge of normal and pathological oriented migration of embryonic cells, and additional cell-guiding activities for chemokines, trophic factors and morphogens.