Enrichment of Mouse Pituitary Thyrotropes from Hypothyroid Mice Using CD90-Based Fluorescence-Activated Cell Sorting

Area of Science:

• Endocrinology
• Cell Biology
• Molecular Biology

Background:

• Thyrotropes are rare anterior pituitary cells (~5%) crucial for thyroid hormone regulation.
• Studying thyrotrope physiology is challenging due to their low abundance.

Purpose of the Study:

• To develop an enhanced protocol for isolating and enriching mouse thyrotropes.
• To facilitate downstream molecular and physiological analyses of thyrotropes.

Main Methods:

• Induction of hypothyroidism to expand thyrotrope populations in adult mice.
• Enzymatic dissociation of anterior pituitary glands followed by fluorescence-activated cell sorting (FACS) using CD90.2.
• Quantification of Tshb mRNA expression for enrichment assessment.

Main Results:

• Isolation of approximately 1.5 × 10^6 CD90.2-positive cells from twelve mice.
• CD90.2-positive cells represented ~25% of pituitary cells with significantly higher Tshb expression.
• Isolated thyrotropes demonstrated viability and responsiveness to triiodothyronine in short-term culture.

Conclusions:

• The described protocol effectively enriches viable mouse thyrotropes.
• This method enables comprehensive molecular, transcriptomic, epigenomic, and physiological studies.
• The enhanced isolation technique overcomes previous limitations in thyrotrope research.