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Wnt signalling in oestrogen-induced lactotroph proliferation
[Thesis]. Manchester, UK: The University of Manchester; 2011.
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Abstract
The University of ManchesterAdam GilesDoctor of Philosophy – PhDWnt signalling in oestrogen-induced lactotroph hyperplasia2011The anterior pituitary gland is the major hormonal regulator in the body. The gland contains five secretory cell types whose emergence during development is defined by a tightly regulated array of transcription factors. In adult life, the gland is plastic with the relative proportions of cells varying depending on physiological context. Tumours of the pituitary gland account for 15% of all intracranial tumours in man and are caused by the selective proliferation of one of the secretory cell types. The majority of these (60%) are prolactinomas which consist of very slowly proliferating lactotroph cells, which produce the hormone prolactin. Pituitary tumours are almost never malignant and do not express common genetic markers for cancer, suggesting endogenous proliferative stimuli in the pituitary are the cause of tumour development.Oestrogen causes lactotroph hyperplasia during pregnancy and increases prolactin secretion. Our group previously showed that Wnt-4 mRNA was upregulated during oestrogen-induced lactotroph hyperplasia in Fischer 344 rats. Wnt molecules are key regulatory proteins controlling differentiation, proliferation and migration in development and adult life. Wnt-4 is involved in the emergence of lactotrophs during development, and has been implicated in pituitary tumour formation. Wnt molecules signal through three pathways. The well studied canonical pathway has been implicated in numerous cancers and centres around gene transcription initiated by translocation of β-Catenin into the nucleus. There are two non-canonical pathways: the Wnt-planar cell polarity (PCP) pathway and the Wnt-calcium pathway which are both poorly understood.In this thesis, the expression of Wnt-4 was studied in the pituitary, and effects of downstream signalling pathways in response to oestrogen were assessed. Wnt-4 was expressed in all secretory cell types of the pituitary, as well as the marginal zone (MZ), a region of the pituitary that may harbour stem cells. Oestrogen upregulated Wnt-4 protein in the somatolactotroph GH3 cell line, though this could not be replicated in primary tissue. A number of approaches (western blotting, immunofluorescence, reporter gene assays and mutant β-Catenin overexpression) were utilised to show that the canonical pathway was not activated in the pituitary. Wnt-4 had a clear inhibitory effect on calcium oscillations in GH3 cells, showing for the first time a non-canonical effect in the pituitary, though the downstream effects are currently unknown. Attempts made to study the activation of the PCP pathway were inconclusive. Efforts focused on the distribution of key structural and regulatory proteins in the anterior pituitary and the MZ. The MZ was characterised by a single layer of cells at the border of the anterior and intermediate lobes of the pituitary, with high expression of E-Cadherin and Sox 9, though no change in distribution was observed with oestrogen treatment. In the anterior lobe, oestrogen treatment decreased N and E-Cadherin expression, which could be an indicator of PCP pathway activation during oestrogen induced-lactotroph hyperplasia. Overall, data suggest that Wnt-4 does not directly cause oestrogen-induced lactotroph proliferation, but is likely to play a role in regulating tissue plasticity in the adult gland, as well as in the pathogenesis of pituitary tumours.