Fig. 5. Upregulated C6st1 mRNA in Schwann cells during an early phase of tissue reorganisation after nerve crush. (A) In situ hybridisation patterns of C6st1 mRNA revealed with DIG-labelled antisense probes on transverse cryosections of normal and post-crush sciatic nerves (at 1, 3, 7, 14 and 28 days as indicated). Arrows in the zoom views (insets) indicate significant C6st1 transcripts in cells circumferential to axons (1 dpc), in irregularly shaped cells profusely distributed about Schwann tubes undergoing reorganisation (3 dpc and 7 dpc). By 14 days post crush, signals were detectable only in the small-diameter Schwann tubes. Signals approach the normal pattern by 28 dpc. Occasional C6st1 transcripts are detectable around blood vessels (arrowheads in normal and 28-day images). The DIG-labelled sense probe yielded negligible hybridisation signal against the background as exemplified with a section of a 14 dpc sciatic nerve. (B) RT-PCR analysis of C6st1 mRNA expression for the profile of change in post-crush sciatic nerves (days 1-28 as indicated) against the expression of GAPDH as an internal reference. Gel bands are shown directly above the corresponding histogram of C6ST: GAPDH intensity ratio. Results are the mean ± s.e.m. of three independent sets of analyses. Bars, 200 µm.