Sympathetic Innervation of Lymph Nodes

Male SD rats were used to trace the sympathetic nerve fiber endings innervating the lymph nodes. After anesthetizing the rats with pentobarbital sodium (20 mg/kg, i.p.), a total volume of 1 μL 30% FR (fluorochrome, Inc., USA) or 1 μL of 15% biotinylated glucan amine (BDA) was injected into the superior cervical sympathetic ganglia. In the experimental group, 25 cases were injected (except injection failure). The control group (12 cases), in order to prove that FR‐labeled fibers are caused by axonal transport rather than simple free diffusion, we injected the same amount of 30% FR into the position of superior cervical ganglion after resection. After 14 days of survival, the rats were anesthetized with pentobarbital sodium (20 mg/kg i.p.) and perfused with 4% paraformaldehyde prepared with 0.1 M phosphate buffer. The 4–5 ipsilateral cervical lymph nodes were fixed with the same fixative overnight and then cryopreserved with 50% sucrose. The slices (25 μm) were cut transversely on a frozen microtome, mounted on gelatin coated slides, and observed under a laser confocal microscope.

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Wu K., Li R., Zhang Y., Liu Y., Wang M., Huang J., Zhu C., Zhang J., Yuan X, & Liu Q. (2023). The discovery of a new type of innervation in lymphoid organs. Physiological Reports, 11(4), e15604.

Publication 2023

Amine Axonal transport Buffer Cervical Diffusion Fiber Fixative Fluorochrome Frozen Gelatin Glucan Laser microscope Lymph nodes Male Microtome Nerve endings Paraformaldehyde Pentobarbital sodium Phosphate Rats Sucrose Superior cervical ganglion

Corresponding Organization : Soochow University

Other organizations : Shanghai Medical College of Fudan University, Second Affiliated Hospital of Soochow University, Nantong University

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Variable analysis

independent variables

Injection of 1 μL 30% FR (fluorochrome, Inc., USA) or 1 μL of 15% biotinylated glucan amine (BDA) into the superior cervical sympathetic ganglia

dependent variables

Tracing of sympathetic nerve fiber endings innervating the lymph nodes

control variables

Male SD rats
Anesthesia with pentobarbital sodium (20 mg/kg, i.p.)
Survival period of 14 days
Perfusion with 4% paraformaldehyde in 0.1 M phosphate buffer
Fixation and cryopreservation of lymph nodes
Sectioning of lymph nodes into 25 μm slices
Mounting of sections on gelatin coated slides
Observation under laser confocal microscope

positive control

Injection of the same amount of 30% FR into the position of superior cervical ganglion after resection

negative control

Control group (12 cases) to prove that FR‐labeled fibers are caused by axonal transport rather than simple free diffusion

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