Characterization of embryonic hypothalamic neurospheres and the role of sex steroids in the developing hypothalamus
Abstract
There is emerging evidence indicating sex steroids may function as neurotrophic factors during brain development. Our objective was to determine whether sex steroids influence murine hypothalamic development during peak neurogenesis. We hypothesized during development hypothalamic neural stem and progenitor cells (NSPCs) respond to hormones to influence proliferation and/or self-renewal. Initially, we characterized the neurosphere assay as an in vitro model system to study hypothalamic NSPCs. We isolated hypothalamic NSPCs and assessed various culturing parameters such as media, density, growth factors, and extrinsic signals. We then utilized this assay to test whether exposure to androgens and estrogens alters their growth. Pharmacological treatment of DHT enhanced the sphere-forming capacity of primary neurospheres. Self-renewal capacity was also increased with physiologically-relevant doses of DHT and E2 exposed only as secondary cultures. Overall, we provide a working protocol for studying embryonic hypothalamic neurospheres and evidence that hypothalamic NSPC behaviour can be modulated by sex steroids.
Description
Keywords
Neuroscience
Citation
Thornton, H. (2017). Characterization of embryonic hypothalamic neurospheres and the role of sex steroids in the developing hypothalamus (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/25274