Intra-axonal Diffusivity Research Articles

Intra-axonal diffusion of [3H]acetylcholine and [3H]gamma-aminobutyric acid in a neurone of Aplysia.

1. [3H]acetylcholine (ACh) or [3H]gamma-aminobutyric acid (GABA) was injected into the cell body of a cholinergic neurone of Aplysia kurodai. 2. [3H]ACh moved down the axon at a maximum speed of 2.5 mm/sec at 20 degrees C. 3. 20 mM-colchicine suppressed this movement, but some passive movement of radioactivity was noted along the axon. 4. Profiles of the passive movement coincided with theoretically obtained diffusion profiles. 5. The diffusion coefficient of ACh in the axoplasm was estimated. It was 3 x 10(-6) cm2/sec at 5 degrees C, 4 X 10(-6) cm2/sec at 15 degrees C and 6.5 x 10(-6) cm2/sec at 30 degrees C. The Q10 was 1.35, the activation energy was about 5 kcal/degrees C. These diffusion coefficients were lower than that of free diffusion of ACh (8 x 10(-6) cm2/sec at 18 degrees C, Fatt, 1954), and assumed to be reasonable, if one considers frictional resistivity of axoplasm in the diffusion of ACh molecules. 6. [3H]GABA diffused similarly to ACh, and the diffusion coefficients agreed with the estimated value when the molecular size differences were taken into account. 7. Both ACh and GABA seemed to diffuse in soluble form or as single molecules in the axoplasm. 8. Intra-axonal diffusion is very effective for short distances.

Observations of HRP labeling following injection through a chronically implanted cannula — A method to avoid diffusion of HRP into injured fibers

One of the limitations of the horseradish peroxidase (HRP) tracer method is the diffusion of HRP into injured axons resulting in unintended labeling of neurons not terminating in the injection area. To overcome this limitation, an experiment was designed to inject the HRP through an implanted cannula after degeneration and healing had taken place. It was shown that implantation of a cannula into the internal capsule significantly decreased the number of labeled axons in the injection site, thus limiting the unintended labeling of neurons from that injection. When injections followed implantation of the cannula by 24 h or more, fibers damaged by the cannula had healed or degenerated sufficiently that intraaxonal diffusion of HRP into those injured fibers did not occur. A significant difference between control (without the cannula) and experimental (with the cannula) injections was observed. Extensive axonal and neuronal labeling following the control injections was seen at the injection site and caudate nucleus, and in the thalamus and parietal cortex, respectively. Experimental injections resulted in sparse axonal and neuronal labeling evident mostly with the larger injections of HRP.