Radioimmunoassay for Lys8, Asn9, neurotensin 8-13: tissue and subcellular distribution of immunoreactivity in chickens.
A sensitive and specific radioimmunoassay (RIA) for Lys8, Asn9, neurotensin 8-13 (LANT-6) has been developed which utilizes 125I-labeled LANT-6 and rabbit antisera raised towards conjugates of synthetic LANT-6 and bovine thyroglobulin. The antiserum described (TG-22) allows the detection of ca 100 fmol of LANT-6 and crossreacts less than 0.01% with chicken or bovine NT. Dose-response relationships for the native (chicken) and synthetic peptides were indistinguishable. Using this assay the distribution of immunoreactive LANT-6 (iLANT-6) through various tissues of the chicken was studied and compared with that of chicken NT (iNT) determined by RIA. Both iNT and ILANT-6 were found primarily in the brain and gastrointestinal tract, however, their regional distributions were found to differ. Subcellular distribution studies in homogenates of chicken brain indicated that both iNT and iLANT-6 were associated with synaptosome-like and vesicle-like particles. In homogenates of small intestine, pancreas and colon iNT and iLANT-6 appeared to be within osmotically sensitive, sedimentable particles. Analyses using high pressure liquid chromatography established that chicken iLANT-6 co-eluted with the synthetic peptide and that similar substances were present in extracts of rat brain and intestine. These results are consistent with "messenger' roles for these peptides.
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007457:Iodine Radioisotopes; D009842:Oligopeptides / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods; D013347:Subcellular Fractions / Q000032:analysis*
D007457:Iodine Radioisotopes; D009842:Oligopeptides; C037243:neurotensin-related hexapeptide