Neural circuits underlying fertility (360G-Wellcome-212242_A_18_Z)

This programme aims to establish in detail the characteristics and in vivo significance of the arcuate (ARNKISS) and rostral periventricular area of the third ventricle (RP3VKISS) kisspeptin neurons in driving the pulsatile and surge patterns of luteinising hormone (LH), respectively. Genetic cFOS-dependent activation and rabies trans-synaptic strategies will enable permanent GFP/mCherry tagging of RP3VKISS neurons activated at the time of the surge, or projecting directly to GnRH neurons, for subsequent electrophysiological and RNAseq analyses. GCaMP-based fiber photometry and GRIN lens miniscopes will be used to evaluate RP3VKISS neuron and population activity. This will be combined with microfluidics delivering pharmacological agents or adeno-associated viruses bearing CRISPR components to interrogate the key factors regulating the activity of RP3VKISS neurons in vivo. Studies aimed at understanding how the ARNKISS neurons synchronise to generate LH pulses will use the same strategies in addition to expansion microscopy and brain slice dual calcium and electrophysiological recordings. Novel strategies for slowing LH pulses in an animal model of polycystic ovary syndrome will be explored. It is expected that the in-depth understanding of these two kisspeptin populations will provide opportunities for developing new therapeutics aimed at the beneficial regulation of fertility in the clinic.