Table 3.

Effect of the simultaneous downregulation of Ror1 and Ror2 by antisense oligonucleotides on neurite elongation in hippocampal neurons

Days in culture	Treatment (oligonucleotide)	Dose (μM)	Primary minor process length (μm)	% Branched primary minor processes	Primary axon length (μm)	Number of branches/axon
1	None	–	34±1	41±4	–	–
1	Sense Ror1+Sense Ror2	50	35±2	39±4	–	–
1	Antisense Ror1+Antisense Ror2	50	27±1*	24±3*	–	–
2	None	-	37±2	57±4	107±5	2.7±0.2
2	Sense Ror1+Sense Ror2	50	31±2	50±4	112±3	2.2±0.2
2	Antisense Ror1+Antisense Ror2	50	26±1*	30±2*	165±8*	1.9±0.2†

E16 hippocampal cultures were plated at 75,000 cells/dish and allowed to develop for 1 (stage II) or 2 (stage III) days. Cells were fixed and stained with a tubulin antibody (clone DMIA). Neuritic processes from randomly selected cells were viewed by phase-contrast microscopy using a digital camera and traced onscreen. Neuritic length was determined using Metamorph Image Analysis Software. Forty cells from three independent experiments were analyzed for each condition. Results are expressed as mean±s.e.m.

^*P<0.01 compared to all controls;

^†P<0.01 compared to untreated controls.