Daytime spikes in dopaminergic activity drive rapid mood-cycling in mice.

Disruptions in circadian rhythms and dopaminergic activity are involved in the pathophysiology of bipolar disorder, though their interaction remains unclear. Moreover, a lack of animal models that display spontaneous cycling between mood states has hindered our mechanistic understanding of mood switching. Here, we find that mice with a mutation in the circadian Clock gene (ClockΔ19) exhibit rapid mood-cycling, with a profound manic-like phenotype emerging during the day following a period of euthymia at night. Mood-cycling coincides with abnormal daytime spikes in ventral tegmental area (VTA) dopaminergic activity, tyrosine hydroxylase (TH) levels and dopamine synthesis. To determine the significance of daytime increases in VTA dopamine activity to manic behaviors, we developed a novel optogenetic stimulation paradigm that produces a sustained increase in dopamine neuronal activity and find that this induces a manic-like behavioral state. Time-dependent dampening of TH activity during the day reverses manic-related behaviors in ClockΔ19 mice. Finally, we show that CLOCK acts as a negative regulator of TH transcription, revealing a novel molecular mechanism underlying cyclic changes in mood-related behavior. Taken together, these studies have identified a mechanistic connection between circadian gene disruption and the precipitation of manic episodes in bipolar disorder.

An important role for cholecystokinin, a CLOCK target gene, in the development and treatment of manic-like behaviors.

Mice with a mutation in the Clock gene (ClockΔ19) have been identified as a model of mania; however, the mechanisms that underlie this phenotype, and the changes in the brain that are necessary for lithium's effectiveness on these mice remain unclear. Here, we find that cholecystokinin (Cck) is a direct transcriptional target of CLOCK and levels of Cck are reduced in the ventral tegmental area (VTA) of ClockΔ19 mice. Selective knockdown of Cck expression via RNA interference in the VTA of wild-type mice produces a manic-like phenotype. Moreover, chronic treatment with lithium restores Cck expression to near wild-type and this increase is necessary for the therapeutic actions of lithium. The decrease in Cck expression in the ClockΔ19 mice appears to be due to a lack of interaction with the histone methyltransferase, MLL1, resulting in decreased histone H3K4me3 and gene transcription, an effect reversed by lithium. Human postmortem tissue from bipolar subjects reveals a similar increase in Cck expression in the VTA with mood stabilizer treatment. These studies identify a key role for Cck in the development and treatment of mania, and describe some of the molecular mechanisms by which lithium may act as an effective antimanic agent.

CCAAT/enhancer binding protein alpha assembles essential cooperating factors in common subnuclear domains.

The transcription factor CCAAT/enhancer binding protein alpha (C/EBP alpha) is the DNA binding subunit of a multiprotein complex that regulates the pituitary-specific GH promoter. C/EBP alpha is absent from the GHFT1-5 pituitary progenitor cell line in which ectopic C/EBP alpha expression leads to activation of the otherwise dormant GH promoter. Transcriptional regulatory complexes are commonly envisaged as assembling from components that evenly diffuse throughout the nucleoplasm. We show that C/EBP alpha, expressed in GHFT1-5 cells as a fusion with color variants of the green fluorescent protein (GFP), concentrated specifically at peri-centromeric chromosomal domains. Although we found the CREB-binding protein (CBP) to activate C/EBP alpha-dependent transcription, CBP was absent from the pericentromeric chromatin. C/EBP alpha expression was accompanied by the translocation of endogenous and ectopically expressed CBP to pericentromeric chromatin. The intranuclear recruitment of CBP required the transcriptional activation domains of C/EBP alpha. C/EBP alpha also caused GFP-tagged TATA binding protein (TBP) to relocate to the Hoechst-stained domains. The altered intranuclear distribution of critical coregulatory factors defines complexes formed upon C/EBP alpha expression. It also identifies an organizational activity, which we label "intranuclear marshalling," that may regulate gene expression by determining the cooperative and antagonistic interactions available at specific nuclear sites.

Altered retinoid signaling in the heads of small eye mouse embryos.

The vitamin A derivative retinoic acid (RA) is necessary for eye development, though its role in signaling within eye tissues is poorly understood. We investigated this question in two transgenic mouse strains carrying a retinoic acid response element (RARE) fused to beta-galactosidase that identify regions of the embryo expressing activated retinoic acid receptors. Retinoid signaling appears in the retina and lens ectoderm of wild-type embryos prior to neural tube closure, when lens induction is under way. To determine if there are interactions between retinoid signaling and the transcription factor Pax-6, also essential for lens development, we examined RARE transgene expression in Small eye (Sey) mice, which carry a Pax-6 mutation. Retinoid signaling in the eye, nose, and forebrain of Sey embryos is decreased, with the most severe effects in the developing lens. In Sey mice the lens anlage cannot respond to exogenous RA after E9, though it is responsive earlier; the retina and other neural ectoderm can respond to RA at any stage. In Sey mice the ability of presumptive lens and retina to produce and/or sequester RA is also decreased, as assayed with a retinoid-reporter cell line. These results implicate retinoid signaling in lens formation and show that RA signaling in the developing eye is dependent upon Pax-6.