Estrogen-dependent development and transcriptome regulation of the lateral septal kisspeptin system
Although hypothalamic kisspeptin neurons have established roles in puberty and reproduction, kisspeptin neurons located in the lateral septum and other extrahypothalamic areas have received less attention. Our goal was to investigate the estrogen-regulated transcripts of KPLS neurons using surgically ovariectomized adult KP-Cre/ZsGreen mice. On post-ovariectomy day 9, the mice were implanted with a single silastic capsule containing either E2 (OVX+E2 group; N=12) or a sunflower oil vehicle (OVX+Veh group; N=9) for a four-day treatment. The brains were fixed using transcardiac perfusion with 0.5% formaldehyde followed by 20% sucrose. The LS was cut into sections using a cryostat, and Laser Capture Microdissection isolated ZsGreen-tagged KP neurons. The OVX+Veh (N=3) and OVX+E2 (N=4) libraries were subjected to single-end sequencing using the Illumina NextSeq500/550 High Output Kit (v2.5; 75 cycles) and an Illumina NextSeq500 instrument.