We quantified tyrosine hydroxylase (TH) expressing TIDA neurons in ageing rats (2 year Bioassay), in order to explore the possibility  that delayed senescence of these neurons may be associated with the development of uterine tumours, as a result of dopaminergic inhibition of prolactin secretion via the hypothalamopituitary axis.

(1) Dual staining for TH RNA (ISH) and protein (IHC)  Formalin Fixed Paraffin Embedded tissue. Subsequent quantification using Image Analysis.

Figure 1: TH RNA by ISH (red) and TH protein IHC (green) dual staining – a more accurate way of measuring dopaminergic neurons

(2)  Microarray and pathways analysis of laser dissected FFPE tissues known to be regulated by the hypothalmo-pituitary axis, to explore signalling pathways (e.g. Estrogen Receptor signalling), that might be associated with uterine carcinogenesis.

The results supported a mode of action consistent with non human relevance (presented at SOT poster session in 2013 and 2014).

(B) – Evaluation of CAR activation and cell proliferation – key events in rat and mouse liver carcinogenesis

In vivo assays for CAR activation and cell proliferation in mouse liver using fluorescence immunostaining

Figure 2: CAR immunostaining  (red) and TOPRO nuclear counterstain  (blue) in phenobarbital treated mouse liver

Figure 3: Anti-BrDU fluorescence immunostaining quantified by automated semi-automated image analysis 

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