The activation of the gonadotropin-releasing hormone (GnRH) neurons from a state of relative quiescence is critical for initiating puberty in mammals. Kisspeptin and its G-protein coupled receptor Gpr54 are essential for puberty, with disruption to either resulting in failed puberty in humans and mice. Robust data from several species indicate that Kiss1 mRNA and/or kisspeptin peptide expression within the hypothalamus increases during pubertal development. Kisspeptin fiber innervation of GnRH neurons and kisspeptin release within the hypothalamus also increase during pubertal development, indicating that there is increased kisspeptinergic drive to GnRH neurons during pubertal development. It is becoming increasingly apparent that gonadal steroids play important roles in the regulation of kisspeptin expression during pubertal development, and in particular, estradiol signaling through estrogen receptor alpha appears to be necessary for these changes to occur. This review focuses on the role that estradiol plays in the regulation of kisspeptin expression during pubertal development.