Figure 4 summarizes innervation status and Table 1 provides group means for all transplant experiments.
These mice and YFP16 wildtypes were used for all transplant experiments involving the SOD1 mutation.
All transplants were performed under general anesthesia (ketamine 24 mg, xylazine 1.3 mg per ml)) For all transplant experiments, the right medial gastrocnemius (MG) muscle was carefully exposed and the tendons of origin and insertion and the MG nerve were severed.
Controls for the transplant experiments included calvarial defects implanted with or without Matrigel™.
For example, transplant experiments of Arnica montana, have revealed that slug herbivory limits the lower elevational range of this subalpine plant (Bruelheide & Scheidel, 1999).
Heterozygous animals were crossed to the C57BL/6-Tg CAG-EGFP 1Osb/J C57BL/6-Tg CAG-EGFP 1Osb/J C57BL/6-Tg CAG-EGFP 1Osb/Jn line), and subsequently backcrossed to produce Lepr db/db animals carrying the eGFP transgene for BM transplant experiments.
For the transplant experiments, Lympholyte M (Cedarlane Laboratories, Ontario, Canada) was used in some cases, according to the manufacturer's instructions, to remove dead cells and red blood cells, and 30 million unlabeled 8.3NOD Scid splenocytes were injected iv per mouse.
For both types of transplant experiment, there was an equivalence of innervation status between transplant and contralateral control muscle pairs.
After 48 hours, LSK cells were selected with puromycin (Sigma,) 2 µg/ml for additional 48 hours and used for coculture or transplant experiments.
For bone marrow transplant experiments, fetal liver from PKBα+/+ and PKBα−/− E15.5 embryos (CD45.2) were dissected and disrupted to single cell suspension by passages through a G25-syringe.
