Abstract
The pituitary is a complex endocrine tissue composed of a number of unique cell types distinguished by the expression and secretion of specific hormones, which in turn control critical components of overall physiology. The basic function of these cells is understood; however, the molecular events involved in their hormonal regulation are not yet fully defined. While previously established cell lines have provided much insight into these regulatory mechanisms, the availability of representative cell lines from each cell lineage is limited, and currently none are derived from adult pituitary. We have therefore used retroviral transfer of SV40 T-antigen to mass immortalize primary pituitary cell culture from an adult mouse. We have generated 19 mixed cell cultures that contain cells from pituitary cell lineages, as determined by RT-PCR analysis and immunocytochemistry for specific hormones. Some lines expressed markers associated with multipotent adult progenitor cells or transit-amplifying cells, including SOX2, nestin, S100, and SOX9. The progenitor lines were exposed to an adenylate cyclase activator, forskolin, over 7 days and were induced to differentiate to a more mature gonadotrope cell, expressing significant levels of α-subunit, LHβ, and FSHβ mRNAs. Additionally, clonal populations of differentiated gonadotropes were exposed to 30 nM gonadotropin-releasing hormone and responded appropriately with a significant increase in α-subunit and LHβ transcription. Further, exposure of the lines to a pulse paradigm of GnRH, in combination with 17β-estradiol and dexamethasone, significantly increased GnRH receptor mRNA levels. This array of adult-derived pituitary cell models will be valuable for both studies of progenitor cell characteristics and modulation, and the molecular analysis of individual pituitary cell lineages.
Highlights
The pituitary gland is known as the master gland because of its central role in governing endocrine activities, together with the hypothalamus
The pituitary is a multifaceted organ that consists of a number of unique cell types controlling many aspects of endocrine physiology [1]
The analysis of the individual cell types and their cell biology is of interest, but the available cell models are limited
Summary
The pituitary gland is known as the master gland because of its central role in governing endocrine activities, together with the hypothalamus. The hypothalamus releases trophic factors that act on specific cell types in the pituitary, stimulating or inhibiting the secretion of pituitary hormones. These pituitary hormones act on peripheral target organs to regulate growth and development, reproduction, metabolism and stress adaptation. Each of the cell lines was derived independently and difficult to compare, due to species differences, tumour-versus SV40 T-antigen transgenically-derived cells, and variation in the temporal isolation of each line These cell lines may not be wholly representative of adult pituitary cells, as many of these lines originated from pituitary tumors at an early developmental stage. Limited cell models from the posterior and intermediate lobes have presently been generated [1]
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