Anterograde Single Labeling System

Tracing techniques in anterograde directions can be used to map the input and output connections at the individual cell levels and imaging and network analysis approach to build mesoscale connectomes for brains. For anterograde tracing, the uptake mechanisms involve the cell soma and/or its dendrites. The tracer material is transported along with the axonal microtubular system to the cell's synaptic terminals.

Anterograde Tracers

Anterograde tracers are used for tracing the anatomical targets of a particular class of projection neurons. Such a technique utilized the radioactively labeled peptide or protein that contributed to the co-development of autoradiographic techniques. The earliest anterograde tracers are radiolabeled amino acids which are injected into neurons and incorporated into the soma, further to transport to axons and terminal processes identified by autoradiography.

Fluorescent dyes became available for anterograde tracing and can be carried actively within the nerve fibers, but the transported substance moves from the cell body towards the axon terminals. The list has since lengthened with introducing new classes of anterograde tracers, including plant lectins, notably Phaseolus Vulgaris leucoagglutinin. More recently, dextran amines have become the most powerful tools for anterograde tracing because they are relatively simple to use. A great variety of detection methods have been developed for them.

Anterograde Tracing Using Single Labeling System

• Anterograde Tracing of Regenerating Axons with Single PHA-L

PHA-L is one of the most widely used anterograde tracers. PHA-L is detected using IHC, revealing detailed cell morphology, including axon terminal branches to the level of terminal boutons. PHA-L requires longer post-injection survival times to achieve transport compared to other tracers. PHA-L can be injected into the spinal cord followed by transplantation of a Schwann cell (SC)-seeded guidance channel into the lesion gap and infusion of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) into the distal spinal cord. Few days after injection, PHA-L labeled axons were visualized through and beyond the SC graft.

Fig.1 BDA anterogradely labeled axons in the CST and RST of the spinal cord. (Wang, 2012)

• Anterograde Tracing of the Corticospinal Tract and Rubrospinal Tract with Single BDA

The use of BDA as a versatile tracer is a common technique for neural pathway tracing in the central nervous system (CNS). BDA can be transported rapidly from the injection site anterogradely to yield excellent neural labeling. BDA can be injected into the motor cortex or red nucleus of normal adult rats and anterogradely transported in the CST or RST of the spinal cord in the two weeks after the injection. BDA labeled axons can be visualized by immunohistochemistry staining or immunofluorescent staining in both cross and longitudinal sections.

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Reference

1. Wang, X.; et al. Anterograde axonal tract tracing. In Animal Models of Acute Neurological Injuries II. Humana Press. 2012, pp. 461-486

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