Adaptation of the Nauta Method for Use on Degenerating Fibers in Rat Cortex and Thalamus

Lesions produced in the cerebral cortex of rats were studied by Nauta's method for degeneration. The brains were perfused with physiological NaCl solution, followed by 10% neutral (CaCO₃) formalin. The brains were removed and stored in the formalin for 2 wk to 1 yr. Experimental modifications of the staining method showed that its sensitivity for fine degenerating fibers could he enhanced by the following changes: (a) omitting 0.05% potassium permanganate; (b) replacing the hydroquinone-oxalic acid mixture with 0.1% pyrogallol. Procedure: (1) frozen sections to water; (2) 0.5% phosphomolybdic acid, 45 min; (3) distilled water, 1 min; (4) 0.1% pyrogallol (aq.), 2 min; (5) distilled water, 3 washes of 1 min each; (6) 1.5% silver nitrate (aq.), 30 min; (7) distilled water, 1 min, (8) Laidlaw's ammoniated silver carbonate, 10110 sec; (9) Nauta's reducer, 1-2 min; (10) distilled water, 1 min; (11) 1.0% Na₂S₂O₃, 2 min; (12) distilled water 3 changes, 1 min each; (13) dehydrate, clear, and cover. This method gave equally good results on degenerating axons in both cortex and thalamus.