references
 
 
 

DCX

Lissencephaly, or otherwise known as smooth brain, is a disease of the human brain development, characterized by incomplete neuronal migration and a smooth cerebral surface. Through genetic research, two mutated proteins were identified, lissencephaly-1 (LS1) and doublecortin (DCX). Both proteins are highly expressed in the developing brain.

DCX is a protein required for normal neuronal migration in the developing cerebral cortex. DCX expression has been found both in the adult rostal migratory stream and in the DG of the hippocampus. A defect in the DCX gene results in epilepsy and mental retardation. DCX is a microtubule-associated phosphoprotein and it appears selectively located in the periphery of the soma with patterns that overlap microtubule distribution (Francis et al., 1999).

DCX has been shown to be expressed in the DG of adult rats in the early differentiation stage, and can be used to identify young, immature neurons (figure 5, Kee et al., 2002). It has been shown that DCX is typically expressed for the first week after the birth of the neuron (Kuhn and Cooper, 2000).
   
 

DCX Protocol

Positive Control Tissue: Young rat hippocampus fixed with 4% paraformaldehyde.

Tissue: Rat Hippocampus

Fixation: 4% Paraformaldehyde in PBS pH 7.2

Primary Antibody: Goat anti-DCX

Ideal Dilution Ratio 1:100-1000

Secondary Antibody: Alexa 488/568 Chicken anti-goat IgG (H+L), (Molecular Probes, Cat# ).

Optimal dilution 1:200

  • Sections were washed with PBS to remove any sodium azide.
  • DCX labelling: sections were washed with PBS, then incubated in primary antibody to DCX (1:500 in 0.3% Triton-X solution, anti-DCX antibody made in Goat, Molec. Probes) for 48 hours at 4 C with gentle shaking.
  • Wash in PBS for 3x5min.
  • The resulting sections were incubated for 2 hours in the dark at room temperature in fluorescent-conjugated secondary antibody (Alexa 568, chicken anti-goat, Chemicon).
  • Wash in PBS for 3x5min.
  • Wash in distilled water 2x5min.
  • Coverslip with mounting medium (Permafluor for fluorecent tissues and Permount for DAB tissues).