Neuropeptidomic Analysis of a Genetically Defined Cell Type in Mouse Brain and Pituitary.
Cell Chem Biol
; 28(1): 105-112.e4, 2021 01 21.
Article
in En
| MEDLINE
| ID: mdl-33217339
ABSTRACT
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Pituitary Gland
/
Neuropeptides
/
Brain
/
Carboxypeptidase H
Limits:
Animals
Language:
En
Journal:
Cell Chem Biol
Year:
2021
Document type:
Article