The sockeye salmon neo-Y chromosome is a fusion between linkage groups orthologous to the coho Y chromosome and the long arm of rainbow trout chromosome 2.

Unlike other Pacific salmon, sockeye salmon (Oncorhynchus nerka) have an X(1)X(2)Y sex chromosome system, with females having a diploid chromosome number of 2n = 58 and males 2n = 57 in all populations examined. To determine the origin of the sockeye Y chromosome, we mapped microsatellite loci from the rainbow trout (O. mykiss; OMY) genetic map, including those found on the Y chromosomes of related species, in kokanee (i.e. non-anadromous sockeye) crosses. Results showed that 3 microsatellite loci from the long arm of rainbow trout chromosome 8 (OMY8q), linked to SEX (the sex-determining locus) in coho salmon (O. kisutch), are also closely linked to SEX in the kokanee crosses. We also found that 3 microsatellite loci from OMY2q are linked to those markers from OMY8q and SEX in kokanee, with both linkage groups fused to form the neo-Y. These results were confirmed by physical mapping of BAC clones containing microsatellite loci from OMY8q and OMY2q to kokanee chromosomes using fluorescence in situ hybridization. The fusion of OMY2q to the ancestral Y may have resolved sexual conflict and, in turn, may have played a large role in the divergence of sockeye from a shared ancestor with coho.

Identification of the sex chromosome pair in chum salmon (Oncorhynchus keta) and pink salmon (Oncorhynchus gorbuscha).

Fluorescence in situ hybridization (FISH) using a probe to the male-specific GH-Y (growth hormone pseudogene) was used to identify the Y chromosome in the karyotypes of chum salmon (Oncorhynchus keta) and pink salmon (Oncorhynchus gorbuscha). The sex chromosome pair is a small acrocentric chromosome pair in chum salmon and the smallest metacentric chromosome pair in pink salmon. Both of these chromosome pairs are morphologically different from the sex chromosome pairs in chinook salmon (Oncorhynchus tshawytscha) and coho salmon (Oncorhynchus kisutch). The 5S rRNA genes are on multiple chromosome pairs including the sex chromosome pair in chum salmon, but at the centromeres of two autosomal metacentric pairs in pink salmon. The sex chromosome pairs and the chromosomal locations of the 5S rDNA appear to be different in all five of the North American Pacific salmon species and rainbow trout. The implications of these results for evolution of sex chromosomes in salmonids are discussed.

Identification of the sex chromosome pair in coho salmon (Oncorhynchus kisutch): lack of conservation of the sex linkage group with chinook salmon (Oncorhynchus tshawytscha).

Fluorescence in situ hybridization (FISH) using a probe to the male-specific GH-Y (growth hormone pseudogene) was used to identify the Y chromosome in coho salmon (Oncorhynchus kisutch). The sex chromosome pair is morphologically similar to chinook salmon (Oncorhynchus tshawytscha) with the GH-Y localized to the small short arm of the largest subtelocentric chromosome pair. FISH experiments with probes containing sex-linked genes in rainbow trout (Oncorhynchus mykiss) (SCAR163) and chinook salmon (Omy7INRA) showed that the coho sex linkage group is different from chinook and rainbow trout and this was confirmed by segregation analysis for the Omy7INRA locus. The telomeric location of the SEX locus, the presence of shared male-specific markers in coho and chinook salmon, and the lack of conservation of sex-linkage groups suggest that transposition of a small male-specific region may have occurred repeatedly in salmonid fishes of the genus Oncorhynchus.

Developmental patterns of mST3GalV mRNA expression in the mouse: in situ hybridization using DIG-labeled RNA probes.

mST3GalV synthesizes ganglioside GM3, the precursor for simple and complex a- and b- series gangliosides, and the expression and regulation of mST3GalV (CMP-NeuAc: lactosylceramide alpha2,3-sialyltransferase) activity is central to the production of almost all gangliosides, a class of glycosphingolipids implicated in variety of cellular processes such as transmembrane signaling, synaptic transmission, specialized membrane domain formation and cell-cell interactions. To understand the developmental expression of mST3GalV in mice, we investigated the spatial and temporal expression of mST3GalV mRNA during the mouse embryogenesis [embryonic (E) days; E9, E11, E13, E15] by in situ hybridization with digoxigenin-labeled RNA probes. All tissues from E9 and E11 were positive for mST3GalV mRNA. On E13, mST3GalV mRNA was expressed in various neural and non-neural tissues. In contrast to these, on E15, the telencephalon and liver produced a strong expression of mST3Gal V which was a quite similar to that of E13. In this stage, mST3GalV mRNA was also expressed in some non-neural tissues. These data indicate that mST3GalV is differently expressed at developmental stages of embryo, and this may be importantly related with regulation of organogenesis in mice.

Natural selection promotes divergence of transferrin among salmonid species.

Transferrin is an iron-binding protein that plays an important role in iron metabolism and resistance to bacterial infection in a variety of organisms. A comparison of transferrin coding sequences from four salmonid species shows that the rate of evolution at nonsynonymous sites is significantly higher than the rate at synonymous sites, suggesting that positive natural selection for new alleles has played an important role in the evolution of transferrin in some salmon species. We hypothesize that the selective agent driving rapid divergence is interactions between host transferrin and the iron-scavenging proteins of pathogenic bacteria.

Low levels of intraspecific variation in the mitochondrial DNA of chum salmon (Oncorhynchus keta).

A total of 798 individuals from 42 different populations of chum salmon (Oncorhynchus keta) were examined for mtDNA variation. Populations were sampled across the geographic range of the species, from mainland Japan around the Pacific Rim to the state of Washington in the United States. The entire D-loop region (approximately 1 kb) was sequenced for 16 individuals from representative populations. Subregions (approximately 200 nucleotides each) of the D-loop reported to be rapidly evolving in salmon were sequenced for another 29 individuals. Only 4 nucleotide variants were detected, and they occurred in only 4 individuals. Four coding regions of the mtDNA genome were also examined using restriction fragment analysis of products amplified via the polymerase chain reaction. Only one, the region coding for NADH dehydrogenase subunits 5 and 6, showed any variation at this level. The restriction enzyme AseI revealed a polymorphism where the frequency of haplotypes was correlated geographically. We surveyed all individuals for this polymorphism and documented a cline in frequency of the haplotypes around the Pacific Rim. There was a significant frequency difference between Japan and 3 other major geographic regions (Russia, Alaska/Yukon, and British Columbia/Washington) for the presence of the 2 haplotypes. This marker may prove useful in the identification of continent-of-origin for individual chum salmon caught in the open ocean.

Nucleotide sequence and tissue distribution of three insulin-like growth factor I prohormones in salmon.

Tissue distribution and potential alternative splicing of insulin-like growth factor I (IGF-I) messenger RNA were studied using reverse transcriptase-polymerase chain reaction (RT-PCR) on RNA from several tissues at various stages of the life cycle of coho salmon (Oncorhynchus kisutch). DNA sequence analysis of RT-PCR products revealed three IGF-I mRNA transcripts, designated Ea-1, Ea-2, and Ea-3, which code for three distinct prohormones, IGF-IA-1, IGF-IA-2, and IGF-IA-3, respectively. The E-domain of proIGF-IA-1 is 35 amino acids long and shares 77% sequence identity with the E-domain of human proIGF-IA, which is also 35 amino acids long. The proIGF-IA-2 and proIGF-IA-3 E-domains are homologous to the proIGF-IA-1 E-domain but contain 27 and 39 amino acid inserts, respectively, between Lys86 and Glu87. In the human IGF-I gene Lys86 is coded by exon 4 and Glu87 is coded by exon 6. This suggests that Ea-2 and Ea-3 transcripts may be the result of alternative splicing during pre-mRNA processing. All three transcripts were readily detectable using a solution hybridization/RNase protection assay. Furthermore, RT-PCR and DNA sequencing analysis indicate the presence of three IGF-I prohormones in another member of the Salmonidae family, the Atlantic salmon (Salmo salar). An analysis of IGF-I and -II E-domains from several vertebrates suggests that certain chemical and physical properties of the molecule are well conserved despite wide variations in primary structure. Ea-1, Ea-2, and Ea-3 transcripts were found in whole embryos, and liver, muscle, and brain of juvenile and adult salmon. At least one IGF-I transcript was found in heart, kidney, testes, ovary, adipose tissue, and spleen of juvenile salmon. These results indicate that IGF-I is expressed during embryonic development of fish, and that most tissues are capable of IGF-I mRNA production. These data also indicate that pre-mRNA transcripts can be alternatively spliced to yield at least three prohormones.