Abstract
Estradiol (E2) induces neurite outgrowth, neuritic spine development and synaptogenesis in ER containing areas of the rat brain. Such changes may modulate E2 and gender effects on cognition, behavior, and reproductive physiology. Unfortunately, most studies to date have been performed in vivo, and hence elucidation of the cellular effects of E2 on neurons has been difficult. For this reason, we have developed an in vitro model using PC12 rat pheochromocyloma cells, which become postmitotic and and extend neurites in response to Nerve Growth Factor (NGF). Wild-type PC12 (CTL1) cells were stably transfected with the vector pCMV-ERα-neo (JNCI 84:580, 1992) in an attempt to overexpress ER (SER8); or with the control vector pCMV-neo (NE09). Exposure of NEO9 cells to 1-100 ng/ml NGF caused them to extend neurites in a dose-dependent fashion similar to CTL1 cells, but without neuritic spines or intercellular connections. Coadministration of 10−10–10−9 M E2 to NEO9 cells yielded no additional effects on neurite frequency, nor induction of neuritic spines nor intercellular connectivity. Exposure of CTL1 cells to NGF led to similar increases in neurite frequency, but coadministration of E2 led to additive increases in neurite frequency, and also to dose-dependent increases in neuritic spine frequency and intercellular connections. SER8 cells demonstrated similar but augmented effects and at lower doses of E2. Preliminary EM studies of intercellular connections in CTL1 and SER8 cells demonstrate synaptic clefts, posl-synaptic densities, synaptic vesicles, and polyribosomal clusters suggestive of synaptogenesis.
These results support the in vivo observations of estrogen effects on neurite outgrowth, spine development, and synaptogenesis, and extend them in vitro. Thus, estrogen appears to activate an inherent neural morphologic program which affects cellular differentiation. Moreover, these cell lines form the basis of an in vitro model for elucidating the mechanisms by which estrogen induces its effects on neuronal differentiation, and will be utilized in future studies of sex hormones and neura organization and activation.
Article PDF
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Luslig, R., Baas, P. ESTROGEN EFFECTS IN NEURAL CELLS IN VITRO: MORPHOLOGIC CHARACTERIZATION OF ESTROGEN RECEPTOR (ER)-TRANSFECTED PC12 CELLS. Pediatr Res 33 (Suppl 5), S32 (1993). https://doi.org/10.1203/00006450-199305001-00173
Issue Date:
DOI: https://doi.org/10.1203/00006450-199305001-00173