MilliporeSigma
  • Improved regeneration after femoral nerve injury in mice lacking functional T- and B-lymphocytes.

Improved regeneration after femoral nerve injury in mice lacking functional T- and B-lymphocytes.

Experimental neurology (2014-06-27)
Ali Mehanna, Emanuela Szpotowicz, Melitta Schachner, Igor Jakovcevski
ABSTRACT

The immune system plays important functional roles in regeneration after injury to the mammalian central and peripheral nervous systems. After damage to the peripheral nerve several types of immune cells, invade the nerve within hours after the injury. To gain insights into the contribution of T- and B-lymphocytes to recovery from injury we used the mouse femoral nerve injury paradigm. RAG2-/- mice lacking mature T- and B-lymphocytes due to deletion of the recombination activating gene 2 were subjected to resection and surgical reconstruction of the femoral nerve, with the wild-type mice of the same inbred genetic background serving as controls. According to single frame motion analyses, RAG2-/- mice showed better motor recovery in comparison to control mice at four and eight weeks after injury. Retrograde tracing of regrown/sprouted axons of spinal motoneurons showed increased numbers of correctly projecting motoneurons in the lumbar spinal cord of RAG2-/- mice compared with controls. Whereas there was no difference in the motoneuron soma size between genotypes, RAG2-/- mice displayed fewer cholinergic and inhibitory synaptic terminals around somata of spinal motoneurons both prior to and after injury, compared with wild-type mice. Extent of myelination of regrown axons in the motor branch of the femoral nerve measured as g-ratio was more extensive in RAG2-/- than in control mice eight weeks after injury. We conclude that activated T- and B-lymphocytes restrict motor recovery after femoral nerve injury, associated with the increased survival of motoneurons and improved remyelination.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Osmium tetroxide solution, 2.5 wt. % in tert-butanol
Sigma-Aldrich
Osmium tetroxide, ReagentPlus®, 99.8%
Sigma-Aldrich
Osmium tetroxide solution, 4 wt. % in H2O
Sigma-Aldrich
2-Methylbutane, ReagentPlus®, ≥99%
Sigma-Aldrich
2-Methylbutane, puriss. p.a., ≥99.5% (GC)
Sigma-Aldrich
Osmium tetroxide, ACS reagent, ≥98.0%
Sigma-Aldrich
2-Methylbutane, anhydrous, ≥99%
Supelco
2-Methylbutane, analytical standard
Sigma-Aldrich
Os EnCat® 40, extent of labeling: 0.3 mmol/g Os loading
Sigma-Aldrich
2-Methylbutane, suitable for HPLC, ≥99.5%
Diazepam for system suitability, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
2-Methylbutane, ReagentPlus®, ≥99%
Sigma-Aldrich
Osmium tetroxide solution, suitable for electron microscopy, 2% in H2O
Sigma-Aldrich
Osmium tetroxide solution, suitable for electron microscopy, 4% in H2O
Sigma-Aldrich
Osmium tetroxide, Sealed ampule.
Diazepam, European Pharmacopoeia (EP) Reference Standard
Supelco
Diazepam solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Anti-SLC32A1 (N-terminal) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Sodium azide, BioUltra, ≥99.5% (T)
Sigma-Aldrich
Sodium azide, purum p.a., ≥99.0% (T)