ABSTRACT
Mutations in transcription factors often exhibit pleiotropic effects related to their complex expression patterns and multiple regulatory targets. One such mutation in the zinc finger homeobox 3 (ZFHX3) transcription factor, short circuit (Sci, Zfhx3Sci/+ ), is associated with significant circadian deficits in mice. However, given evidence of its retinal expression, we set out to establish the effects of the mutation on retinal function using molecular, cellular, behavioral and electrophysiological measures. Immunohistochemistry confirms the expression of ZFHX3 in multiple retinal cell types, including GABAergic amacrine cells and retinal ganglion cells including intrinsically photosensitive retinal ganglion cells (ipRGCs). Zfhx3Sci/+ mutants display reduced light responsiveness in locomotor activity and circadian entrainment, relatively normal electroretinogram and optomotor responses but exhibit an unexpected pupillary reflex phenotype with markedly increased sensitivity. Furthermore, multiple electrode array recordings of Zfhx3Sci/+ retina show an increased sensitivity of ipRGC light responses.
Subject(s)
Circadian Rhythm/physiology , Homeodomain Proteins/metabolism , Retina/metabolism , Amacrine Cells/metabolism , Animals , Light , Locomotion/physiology , Male , Mice , Mice, Inbred C57BL , Photic Stimulation/methods , Retinal Ganglion Cells/metabolism , Vision, Ocular/physiologySubject(s)
Karyopherins/genetics , Nephrotic Syndrome/genetics , Nuclear Pore Complex Proteins/genetics , Animals , Female , Humans , MaleSubject(s)
Blood Pressure , Endothelium, Vascular/physiology , Homeostasis , Myelin Proteins/physiology , Sphingolipids/metabolism , Animals , Humans , MaleSubject(s)
Body Mass Index , Kidney Diseases/epidemiology , Kidney/physiology , Obesity/complications , HumansABSTRACT
We identified a dominant missense mutation in the SCN transcription factor Zfhx3, termed short circuit (Zfhx3(Sci)), which accelerates circadian locomotor rhythms in mice. ZFHX3 regulates transcription via direct interaction with predicted AT motifs in target genes. The mutant protein has a decreased ability to activate consensus AT motifs in vitro. Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed. Moreover, mutant ZFHX3 had a decreased ability to activate AT motifs in the promoters of these neuropeptide genes. Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices. In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.
