Journal: PLoS ONE
Article Title: The expression of prosaposin and its receptors, GRP37 and GPR37L1, are increased in the developing dorsal root ganglion
Figure Lengend Snippet: (a–f) Immunofluorescence light micrographs of the rat DRG from 3 days to 8 weeks after birth, stained with anti-PSAP (red), anti-sortilin (Solt, green), and DAPI. Weak sortilin-IR was first observed at postnatal day 7 in satellite cells (arrows in b). After postnatal week 2, the number of sortilin-positive cells increased, peaking at postnatal week 4 (arrows in d) and decreasing after postnatal week 8 (f). (g–o) Higher magnification immunofluorescence light micrographs of the rat DRG at 8 weeks after birth stained with antibodies against PSAP (red), sortilin (green), and DAPI (blue). (g, j) Both the outer and inner cell surface of satellite cells were clearly outlined by sortilin, while only the inner surface of satellite cells was outlined with fine granular PSAP (white arrowheads in i, l, and o), and not the outer cell surface (black arrowheads in i, l). (m) Rare case of a thin satellite cell (S) between two neurons (N), indicating clear colocalization of sortilin (n) and PSAP (o). (p–x) The relationships of PSAP and its receptors (GPR37, GPR37L1) with sortilin was studied by double immunofluorescence of the DRG at postnatal week 4. Sortilin-IR was localized in satellite cells, especially near the boundary to nerve cells (q, t, and w), while PSAP-IR (r) and GPR37-IR (u) were dispersed in the cytoplasm of satellite cells. Only GPR37L1 clearly colocalized with sortilin (w, x). The numbers on the bars indicate the scale in μm.
Article Snippet: Following antigen retrieval, sections were incubated in blocking solution containing 5% NSS, 5% BSA, and 0.25% carrageenan in PBS for 2 h. The sections were processed for immunohistochemistry with anti-PSAP (IM-1), anti-GPR37 or anti-GPR37L, and anti-sortilin (Abcam, Tokyo, Japan) antibodies at a concentration of 1 μg/ml overnight at 4°C.
Techniques: Immunofluorescence, Staining