The Homeodomain Transcription Factors Vax1 and Six6 Are Required for SCN Development and Function

The brain's primary circadian pacemaker, the suprachiasmatic nucleus (SCN), is required to translate day-length and circadian rhythms into neuronal, hormonal, and behavioral rhythms. Here, we identify the homeodomain transcription factor ventral anterior homeobox 1 (Vax1) as required for SCN development, vasoactive intestinal peptide expression, and SCN output. Previous work has shown that VAX1 is required for gonadotropin-releasing hormone (GnRH/LHRH) neuron development, a neuronal population controlling reproductive status. Surprisingly, the ectopic expression of a Gnrh-Cre allele (Gnrh^cre) in the SCN confirmed the requirement of both VAX1 (Vax1^flox/flox:Gnrh^cre, Vax1^Gnrh-cre) and sine oculis homeobox protein 6 (Six6^flox/flox:Gnrh^cre, Six6^Gnrh-cre) in SCN function in adulthood. To dissociate the role of Vax1 and Six6 in GnRH neuron and SCN function, we used another Gnrh-cre allele that targets GnRH neurons, but not the SCN (Lhrh^cre). Both Six6^Lhrh-cre and Vax1^Lhrh-cre were infertile, and in contrast to Vax1^Gnrh-cre and Six6^Gnrh-cre mice, Six6^Lhrh-cre and Vax1^Lhrh-cre had normal circadian behavior. Unexpectedly, ~ 1/4 of the Six6^Gnrh-cre mice were unable to entrain to light, showing that ectopic expression of Gnrh^cre impaired function of the retino-hypothalamic tract that relays light information to the brain. This study identifies VAX1, and confirms SIX6, as transcription factors required for SCN development and function and demonstrates the importance of understanding how ectopic CRE expression can impact the results.