Comprehensive Workflow for Analyzing Peptide, Steroid, and Protein Components in Pituitary Samples. (A) Overall sample processing workflow from tissue homogenate to specific samples for corresponding analyses. (B) Chromatogram and mass spectrometry results for CGA and LHB) peptides selected for targeted proteome analysis. (C) Chromatogram and mass spectrometry results for selected peptide and steroid analyses, with AVP, GNRH3, and testosterone serving as model compounds among all analytes. (D) 93 pituitary proteins (5.1%) that were identified with both peptidomics and trypsin-digested proteomics. Also shown are the numbers of other peptides (75; 4.1%) and proteins (1666; 90.8%) identified.

Pituitary proteome and peptidome comparison. Neurohormones identified by proteomics in a single zebrafish that regulate pituitary hormone release (top graph). Neuroendocrine proteins identified by peptidome analysis in a single zebrafish pituitary (bottom graph). For analysis and ease of comparison, we followed nomenclature and abbreviations conventions of HUGO (https://www.genenames.org/): ADM2, adrenomedullin 2; AGT, angiotensinogen; AVP, arginine vasopressin; CRH, corticotropin releasing hormone; FSTL1, follistatin like 1; GAL, galanin and GMAP prepropeptide; GH1, growth hormone 1; LHB, luteinizing hormone subunit beta; NMU, neuromedin U; NPB, neuropeptide B; OXT, oxytocin/neurophysin I prepropeptide; PGRMC1, progesterone receptor membrane component 1; PMCH, pro-melanin concentrating hormone; POMC, proopiomelanocortin; PRL, prolactin; SCG2, secretogranin II; SCG3, secretogranin III; SCG5, secretogranin V; SCP2, sterol carrier protein 2; SST, somatostatin; SMTL, somatolactin; TSHB, thyroid stimulating hormone subunit beta; VIP, vasoactive intestinal peptide.