Cell Biology JoVE (Open Access) Citable · DOI

Application of Impermeable Barriers Combined with Candidate Factor Soaked Beads to Study Inductive Signals in the Chick

Name: Application of Impermeable Barriers Combined with Candidate Factor Soaked Beads to Study Inductive Signals in the Chick
Uploaded: 2026-06-04
Duration: 8 min 4 s
Description: The chick embryo provides a superb vertebrate model that can be used to dissect developmental questions in a direct way. Its accessibility and robustness ...

DOI: 10.3791/54618-v

What you'll learn

✓ Construct impermeable aluminum foil barriers to block tissue induction
✓ Apply candidate factor-soaked beads to rescue developmental signaling
✓ Analyze gene expression changes following barrier and bead manipulation
✓ Identify retinoic acid as essential inductive signal for limb outgrowth

Protocol

Protocols using impermeable barriers to block induction events between tissues of the main body axis and flank in the chick embryo required for limb formation are described. Beads soaked in candidate inductive signals are used to overcome the effect of barrier placement and analysis of gene expression confirms this.

Difficulty

advanced

Total time

~5–7 days (egg incubation + surgical manipulation + gene analysis)

Model organism

Chicken (Gallus gallus domesticus) embryo

Biosafety

BSL-1

Steps

Create impermeable barriers from aluminum foil

Prepare and shape aluminum foil barriers to block tissue-to-tissue communication. These barriers will be inserted between specific embryonic tissues to prevent inductive signals.

▶ 00:51

Window incubated chicken eggs for access

Create observation windows in the eggshell to visualize and access the developing embryo while maintaining sterility and viability during incubation.

▶ 02:08

Insert barrier and place factor-soaked beads at stage 15

Position impermeable barriers between body axis and flank at stage 15 embryonic development, then apply candidate factor-soaked beads to test rescue of limb bud initiation.

▶ 03:35

Block induction at stage 10 and assess outcomes

Apply barriers at earlier stage 10 to prevent leg bud induction and initiation, then use beads to determine if candidate factors restore developmental processes.

▶ 05:20

Analyze gene expression to confirm retinoic acid role

Extract embryonic tissues and perform gene expression analysis to validate that retinoic acid from somites is essential for limb bud outgrowth.

▶ 06:34

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