SUMMARYThe myosuppressin (MS) gene was cloned from a central nervous system (CNS) cDNA library of the hematophagous insect Rhodnius prolixus and is predicted to contain two introns and three exons. The mRNA transcribed from the myosuppressin gene encodes an 88 amino acid prepropeptide, which results in a mature decapeptide after post-translational modification. When compared with the myosuppressins isolated from other insects, the R. prolixus myosuppressin has a unique amino acid sequence (pQDIDHVFMRFamide), with isoleucine (I) in position 3 and methionine (M) in position 8. Reverse transcriptase (RT)-PCR shows that Rhopr-MS is expressed in the CNS and posterior midgut in R. prolixus and immunohistochemistry suggests that an RFamide-like peptide is present in endocrine-like cells in the midgut. Physiological assays using Rhopr-MS indicate that, despite the unusual M at position 8, it still retains myoinhibitory activity, inhibiting the frequency and reducing the amplitude of contractions in the anterior midgut and hindgut, and decreasing heart rate.

Severe appetite and weight loss define the eating disorder anorexia nervosa, and can also accompany the progression of some neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS). Although acute loss of hypothalamic neurons that produce appetite-stimulating neuropeptide Y (Npy) and agouti-related peptide (Agrp) in adult mice or in mice homozygous for the anorexia (anx) mutation causes aphagia, our understanding of the factors that help maintain appetite regulatory circuitry is limited. Here we identify a mutation (C19T) that converts an arginine to a tryptophan (R7W) in the TYRO3 protein tyrosine kinase 3 (Tyro3) gene, which resides within the anx critical interval, as contributing to the severity of anx phenotypes. Our observation that, like Tyro3−/− mice, anx/anx mice exhibit abnormal secondary platelet aggregation suggested that the C19T Tyro3 variant might have functional consequences. Tyro3 is expressed in the hypothalamus and other brain regions affected by the anx mutation, and its mRNA localization appeared abnormal in anx/anx brains by postnatal day 19 (P19). The presence of wild-type Tyro3 transgenes, but not an R7W-Tyro3 transgene, doubled the weight and lifespans of anx/anx mice and near-normal numbers of hypothalamic Npy-expressing neurons were present in Tyro3-transgenic anx/anx mice at P19. Although no differences in R7W-Tyro3 signal sequence function or protein localization were discernible in vitro, distribution of R7W-Tyro3 protein differed from that of Tyro3 protein in the cerebellum of transgenic wild-type mice. Thus, R7W-Tyro3 protein localization deficits are only detectable in vivo. Further analyses revealed that the C19T Tyro3 mutation is present in a few other mouse strains, and hence is not the causative anx mutation, but rather an anx modifier. Our work shows that Tyro3 has prosurvival roles in the appetite regulatory circuitry and could also provide useful insights towards the development of interventions targeting detrimental weight loss.