Two precursors to Chinook salmon (Onco-rhynchus tshawytscha) melanin-concentrating hormone, an important factor in teleosts involved in the control of skin pigmentation and stress responsiveness, have been identified from DNA sequence analysis. Both precursors encode proteins of 132 amino acids and they share 107/132 amino acid identities. The biologically active 17-residue peptide is located at the C terminus of both precursors and can be liberated by proteolytic cleavage following two adjacent arginine residues. Additional putative proteolytic processing sites are located within the two precursors. Northern analysis demonstrated an intense hybridization signal of 750 nucleotides in the hypothalamus.Immunocytochemical studies as well as in situ hybridization analyses identify intensely staining cell bodies in the hypothalamus in the area of the lateral tuberal nucleus.Regulation of skin pigmentation and stress response in amphibians and teleosts has been the source of considerable research over the past few decades. Two types of control over melanophore activity have been demonstrated: neuronal control via monoamines and endocrine control (1-9). Significant effort has been directed at detecting a factor(s) that exerts an inhibitory effect on melanocyte-stimulating hormone (MSH) (1-3, 7, 9-13). Kawauchi and collaborators (12) successfully isolated and characterized such a factor from salmon pituitary in the form of a 17-amino acid peptide named melanin-concentrating hormone (MCH). Synthetic versions of MCH (11,(14)(15)(16)(17) have been shown to be potent in causing the melanophores of teleosts to concentrate their pigment granules (blanching), as well as inhibiting the release of MSH (2,11,12,18,19 Immunocytochemistry and in Situ Colocalization. Tissue sections were prepared as described above. Vanadyl ribonucleoside complex (2 mM) was added to all antibodies to inhibit RNase activity. After incubation with diaminobenzidine, sections were washed in distilled water, dehydrated to 100% ethanol, and air-dried. All sections were prehybridized as described and hybridized with either the MCH RNA probe or oligonucleotides at the appropriate temperature for 24 hr.