Neuropeptidomic Analysis of a Genetically Defined Cell Type in Mouse Brain and Pituitary

Autor(es)

Fricker, Lloyd D. ^[1] ; Tashima, Alexandre K. ^[2] ; Fakira, Amanda K. ^{[3, 4]} ; Hochgeschwender, Ute ^{[5, 6]} ; Wetsel, William C. ^{[7, 8, 9]} ; Devi, Lakshmi A. ^[10]

Afiliação do(s) autor(es)

^[1] Albert Einstein Coll Med, Dept Mol Pharmacol, 1300 Morris Pk Ave F248, Bronx, NY 10461 - USA

^[2] Univ Fed Sao Paulo, Dept Biochem, Escola Paulista Med, BR-04023901 Sao Paulo, SP - Brazil

^[3] Icahn Sch Med Mt Sinai, Dept Pharmacol Sci, New York, NY 10029 - USA

^[4] Rowan Univ, Dept Biomed Sci, Cooper Med Sch, Camden, NJ - USA

^[5] Duke Univ, Med Ctr, Neurotransgen Lab, Dept Neurobiol, Durham, NC 27710 - USA

^[6] Cent Michigan Univ, Coll Med, Neurosci Program, Mt Pleasant, MI 48859 - USA

^[7] Duke Univ, Med Ctr, Dept Cell Biol, Durham, NC 27710 - USA

^[8] Duke Univ, Med Ctr, Dept Psychiat & Behav Sci, Durham, NC 27710 - USA

^[9] Duke Univ, Med Ctr, Dept Neurobiol, Durham, NC 27710 - USA

^[10] Icahn Sch Med Mt Sinai, Dept Pharmacol Sci Neurosci & Psychiat, New York, NY 10029 - USA

• Tipo de documento: Artigo Científico
• Fonte: Cell Chemical Biology; v. 28, n. 1, p. 105+, JAN 21 2021.
• Citações Web of Science: 0

Resumo

Neuropeptides and peptide hormones are important cell-cell signaling molecules that mediate many physiological processes. Unlike classic neurotransmitters, peptides undergo cell-type-specific post-translational modifications that affect their biological activity, To enable the identification of the peptide repertoire of a genetically defined cell type, we generated mice with a conditional disruption of the gene for carboxypeptidase E (Cpe), an essential neuropeptide-processing enzyme. The loss of Cpe leads to accumulation of neuropeptide precursors containing C-terminal basic residues, which serve as tags for affinity purification. The purified peptides are subsequently identified using quantitative peptidomics, thereby revealing the specific forms of neuropeptides in cells with the disrupted Cpe gene. To validate the method, we used mice expressing Cre recombinase under the proopiomelanocortin (Pomc) promoter and analyzed hypothalamic and pituitary extracts, detecting peptides derived from proopiomelanocortin (as expected) and also proSAAS in POMC neurons. This technique enables the analyses of specific forms of peptides in any Cre-expressing cell type. (AU)

• Processo FAPESP: 17/20106-9 - Peptidômica dos venenos de serpentes e aranhas brasileiras
• Beneficiário: Alexandre Keiji Tashima
• Modalidade de apoio: Auxílio à Pesquisa - Regular