SCO-spondin and RF-GlyI are two designations for cDNAs strongly expressed in the bovine subcommissural organ (SCO), characterized, respectively, in 1996 and 1998 by two different research groups. Because both cDNAs were partial sequences and exhibited close similarities in their nucleotide and deduced amino acid sequences, it was thought that they might be part of the same encoding sequence. To find out, we performed 3'RACE using a SCO-spondin-specific upstream primer. From the RT-PCR product generated and by nested PCR techniques, we amplified both SCO-spondin and RF-GlyI specific products with the expected length. Also, probes generated from both PCR products hybridized to the same major 14 kb transcript in Northern blot analyses, clearly showing that SCO-spondin and RF-GlyI cDNAs do belong to the same encoding sequence. In addition, we amplified, cloned, and sequenced a PCR product of 3 kb spanning both the known SCO-spondin and RF-GlyI sequences. The deduced amino acid sequence contains nine thrombospondin type 1 repeats that alternate with sequences sharing similarities with the D-domain of von Willebrand factor. Taken together, these findings show that SCO-spondin and RF-GlyI are two designations of the same gene encoding proteins secreted by the bovine SCO and forming Reissner's fiber. In addition, compared to the sequence provided by Nualart et al. (1998), we extended the reading frame and identified new conserved domains in the 3' end of SCO-spondin. The putative function of SCO-spondin on axonal pathfinding is discussed regarding the presence of a great number of thrombospondin type 1 repeats.
Copyright 2000 Wiley-Liss, Inc.