Publication date: Available online 27 March 2018
Source:Journal of Neuroscience Methods
Author(s): Edith Segura-Anaya, Rommel Flores-Miranda, Alejandro Martínez-Gómez, Myrna A.R. Dent
BackgroundThe Golgi silver method has been widely used in neuroscience for the study of normal and pathological morphology of neurons. The method has been steadily improved and Bielschowsky's silver staining method (BSSM) is widely used in various pathological conditions, like Alzheimer's disease.New methodIn this work, teased sciatic nerves were silver impregnated using BSSM. We also developed simultaneous staining by silver impregnation and single- or double-immunofluorescence of the same section in teased nerve preparations. We immunostained against non-myelinating Schwann cells and different myelinating Schwann cell domains.ResultsBSSM teased nerves show a strong staining of axons (black) and a gold-brown staining of non-myelin Schwann cells and compact myelin. We were also able to stain by immunofluorescence these BSSM teased nerves with specific molecular markers against non-myelinating Schwann cells and different myelinating Schwann cell domains such as the Schmidt-Lanterman incisures, paranodal regions or compact myelin.Comparison with existing methods and ConclusionsIn peripheral nerves, several silver impregnation methods have been used to stain nerves in paraffin sections, but not in teased nerves to enable the assessment of isolated nerve fibers. In conclusion, BSSM gives accurate information of nerve morphology and combining the procedure with immunofluorescence it would be very useful to study the molecular nerve domain organization of the nerve fibers, and to study the molecular pathology of axon degeneration, or myelin disorders, or of any peripheral neuropathy, also to study demyelination diseases in the central nervous system.
Graphical abstract
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