Related Experiment Videos
Rat prolactin synthesis by lactating mammary epithelial cells
M Lkhider1, S Delpal, F Le Provost
1Université Chouaib Doukkali, Faculté des Sciences, El Jadida, Morocco.
FEBS Letters
|January 20, 1997
Summary
Lactating mammary tissue synthesizes prolactin (PRL). Researchers confirmed this by tracking radioactive PRL forms and observing protein synthesis in mammary cells, demonstrating local PRL production.
Area of Science:
- Endocrinology
- Cell Biology
- Biochemistry
Background:
- Previous suggestions indicated mammary cells might produce prolactin (PRL).
- This study aimed to experimentally verify the endogenous synthesis of PRL within mammary tissue.
Purpose of the Study:
- To investigate and confirm the hypothesis that mammary cells can synthesize prolactin (PRL).
- To elucidate the cellular mechanisms and sites of PRL production in lactating mammary tissue.
Main Methods:
- Incubation of mammary tissue with [35S]methionine-cysteine.
- Analysis using SDS-PAGE, immunoblotting, autoradiography, and two-dimensional electrophoresis.
- Immunogold electron microscopy to visualize PRL synthesis and secretion pathways with and without cycloheximide.
Main Results:
- Radioactive forms of immunoreactive prolactin (PRL) with molecular weights of 14, 23, 25, 32, and 36 kDa were detected.
- Two-dimensional electrophoresis identified radioactive and immunoreactive PRL spots around 25 kDa and at high molecular weights.
- Cycloheximide treatment significantly reduced labeling in organelles associated with synthesis and secretion, confirming active protein production.
Conclusions:
- Lactating mammary tissue possesses the capability to synthesize prolactin (PRL).
- The findings support the endogenous production of PRL within the mammary gland itself.