Materials

The Peripheral Neuropathy Solution

The Peripheral Neuropathy Solution

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1. Phosphate-buffered saline (PBS; 10X concentrated, pH 7.2) /L: 2.1 g KH2PO4, 90 g NaCl, 7.26 g Na2HPO47H2O. Autoclaved.

2. Paraformaldehyde (Fluka, Gillingham, UK).

3. Hydrogen peroxide (30% w/w; Sigma, replace every 4 mo).

5. Sodium azide (Sigma).

6. Basal medium (Eagle) containing 25 mM HEPES (Life Technologies, Paisley, Scotland).

7. Calf serum (Life Technologies).

8. Normal goat or donkey sera (Sigma).

Fig. 1. (A) Whole-stage 19 chick embryo stained with an MAb against the cell-adhesion molecule L1/Ng-CAM, which is expressed on a subset of axons in the nervous system. Cranial (arrowheads) and dorsal root (arrows) ganglia of the peripheral nervous system are visible. (B) The hindbrain dissected from the same embryo and mounted flat to visualize axons within hindbrain segments (rhombomeres) and accumulating of axons at exit points (arrows) in rhombomeres 2 and 4 and in interrhombomeric boundaries (arrowheads) are indicated.

Fig. 1. (A) Whole-stage 19 chick embryo stained with an MAb against the cell-adhesion molecule L1/Ng-CAM, which is expressed on a subset of axons in the nervous system. Cranial (arrowheads) and dorsal root (arrows) ganglia of the peripheral nervous system are visible. (B) The hindbrain dissected from the same embryo and mounted flat to visualize axons within hindbrain segments (rhombomeres) and accumulating of axons at exit points (arrows) in rhombomeres 2 and 4 and in interrhombomeric boundaries (arrowheads) are indicated.

9. Peroxidase-conjugated secondary antibodies (Jackson Laboratories).

11. Diaminobenzidine (DAB; 10 mg tablets from Sigma).

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