GSBS Student Publications

Title

Induction of the neurotensin (NT) gene in PC12 cells gives rise to NT precursor (approximately 88%), NT(3-13)-like peptide (approximately 10%), and NT (approximately 2%)

GSBS Program

Biochemistry & Molecular Pharmacology

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Physiology; Department of Molecular Genetics and Microbiology

Date

9-1-1993

Document Type

Article

Medical Subject Headings

Animals; Culture Media; Gene Expression Regulation; Membrane Potentials; Neurotensin; PC12 Cells; Peptide Fragments; Potassium; Precipitin Tests; Protein Precursors

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Neurotensin (NT) is coexpressed with catecholamines in sympathetic neurons and adrenal chromaffin cells. A pheochromocytoma PC12 cell line can also be induced to express the NT gene and produce immunoreactive NT. In the present study, NT mRNA was quantified under various hormonal conditions and NT precursor synthesis rates were determined by pulse labeling and immunoprecipitation. In addition, NT precursor and NT-related products were measured using RIA and were characterized using HPLC and Sephadex chromatography. Neurotensin mRNA, NT precursor synthesis, and NT precursor/product levels were correlated. Surprisingly, NT appeared to be a minor product, both in cells and media: NT precursor (approximately 88%), NT(3-13)-like peptide (approximately 10%), and NT (approximately 2%). Neurotensin added to cultures was not converted to NT(3-13). Treatment of cells with 60 mM KCl or various secretagogues induced Ca(2+)-dependent release of NT precursor, NT(3-13), and NT in proportion to their cellular contents. These results suggest a) that NT precursor processing in induced PC12 cells was much slower than NT precursor synthesis, b) that NT(3-13) was a major product and NT a minor one, and c) that NT precursor and its products were stored within secretory vesicles.

Rights and Permissions

Citation: Peptides. 1993 Sep-Oct;14(5):991-9.

Related Resources

Link to article in PubMed

Journal Title

Peptides

PubMed ID

8284275