Article Figures & Data
Figures
- Fig. 1.
Southern blot. Five micrograms of DNA extracted from female (a) and male (b) tissue were digested with EcoRI, electrophoresed in agarose gel (1%), blotted on a nylon membrane, and hybridized with the 145SC5 probe, which detected a strong signal of 1.5 kb in the male DNA. The 2.2 kb band detected by hybridization with a probe for the prion protein gene demonstrates similar amounts of DNA on each lane.
- Fig. 2.
a, b, Numerous macrophages are present within the transplanted male sciatic nerve, as detected by immunocytochemistry for the Mac-3 (DPT 5) (a) or F4/80 (DPT 20) (b) antigens.c–h, Detection of apoptotic macrophages within the transplanted male donor nerve. c, Morphologically, cells with the typical hallmarks of apoptosis (arrows) such as chromatin condensation and margination and apoptotic bodies are present (H&E). d, Immunocytochemistry for Mac-3 shows an apoptotic macrophage (arrow) with a condensed and fragmented nucleus. e, Semithin section of sciatic nerve 10 d after transection. Myelin debris-containing macrophages with dark, condensed nuclei (arrows) typical for apoptosis are present. f, Detection of DNA fragmentation with thein situ tailing technique. Degenerating cells (arrows) are present within the transplanted nerve.g, Double staining for IST and F4/80 identifies a macrophage with DNA fragmentation (arrow).h, Immunocytochemistry for CM-1. CM-1-positive (arrow) and CM-1-negative (arrowhead) cells with the typical morphology of apoptosis. Scale bars, 25 μm.
- Fig. 3.
Quantitative data of invading (a, b) and apoptotic (c, d) macrophages in transplant (a, c) and recipient nerves (b, d). The open square represents cell numbers in the degenerating control nerve. *Statistically significant (p < 0.05) versus the total number of endoneurial macrophages at DPT 2 and 5 and DPT 20 as well as the relative number of apoptotic macrophages at the same time points. **Statistically significant (p < 0.0005) versus the number of endoneurial macrophages at all other time points.
- Fig. 4.
Double immunofluorescence for CM-1 (green) and Mac-3 (red). The double-labeled macrophage is clearly detected.
- Fig. 5.
Detection of Y chromosome-positive cells. a, H&E stain showing the highly cellular transplanted nerve at the left margin and several fascicles of the host nerve at the right margin. b, The same nerve as shown ina stained for the Y-chromosome probe 145SC5. There are numerous positive cells within the transplanted nerve and the surrounding perineurial tissue. c, High magnification of the grafted nerve showing numerous large, round Y chromosome-positive cells with macrophage morphology. d, Y chromosome-positive cells are also present in the surrounding perineurial tissue of the transplanted nerve. e, Few Y chromosome-positive cells are detected in the female host nerve.f–h, Y chromosome-positive cells (f) closely attached to an endothelial cell at the luminal surface (arrow) of a perineurial blood vessel, (g) in a para-aortal lymph node and (h) in the spleen. Scale bars:a, b, 100 μm; c–h, 25 μm.
Tables
DPT Graft nerves Recipient nerves Perineurial tissue Regional lymph nodes1-a Paraaortal lymph nodes Spleen 2 (n = 3) +++/+++/+++ +/−/− +++/+++/+++ ++/−/− +++/+++/+ +++/+++/++ 5 (n = 3) +++/+++/+++ +/−/− +++/+++/+++ −/−/− +/+/+ +++/−/+ 10 (n = 3) +++/+++/+++ +/−/− +++/+++/+++ −/−/− ++/++/+ +++/++/− 20 (n = 3) +++/+++/+++ −/−/− +++/+++/+++ +++/−/− +/+++/++ +++/−/+ 40 (n = 2) +++/+++ −/− +++/+++ +/− +/+ +++/− -
DPT, Days after transplantation; +++, more than 10 145SC5 positive cells, ++, between 5 and 10 positive cells, +, between 1 and 4 positive cells.
-
↵F1-a Regional lymph nodes were only seen in three animals with the presence of 145SC5-positive cells in each of them.
-
In this issue
