Transgenic expression of Cre recombinase from the tyrosine hydroxylase locus

Lindeberg, J̇onas; Usoskin, Dmitry; Bengtsson, Henrik; Gustafsson, Anna; Kylberg, Annika; Söderström, Stine; Ebendal, Ted

doi:10.1002/gene.20065

Cited by 197 publications

(219 citation statements)

References 32 publications

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“…2A). We genetically targeted optogenetic transgenes to LC neurons by stereotaxically injecting a Cre recombinase-dependent adeno-associated virus (AAV) into knock-in mice selectively expressing Cre in tyrosine hydroxylase neurons (TH::IRES-Cre mice) (26). AAV vectors contained either eNpHR3.0-eYFP or eYFP and were previously shown to specifically deliver transgenes to >98% of tyrosine hydroxylase-positive neurons in the LC (24).…”

Section: Resultsmentioning

confidence: 99%

Mechanism for Hypocretin-mediated sleep-to-wake transitions

Carter

Brill

Bonnavion³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

247

203

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Current models of sleep/wake regulation posit that Hypocretin (Hcrt)-expressing neurons in the lateral hypothalamus promote and stabilize wakefulness by projecting to subcortical arousal centers. However, the critical downstream effectors of Hcrt neurons are unknown. Here we use optogenetic, pharmacological, and computational tools to investigate the functional connectivity between Hcrt neurons and downstream noradrenergic neurons in the locus coeruleus (LC) during nonrapid eye movement (NREM) sleep. We found that photoinhibiting LC neurons during Hcrt stimulation blocked Hcrt-mediated sleep-to-wake transitions. In contrast, when LC neurons were optically stimulated to increase membrane excitability, concomitant photostimulation of Hcrt neurons significantly increased the probability of sleep-to-wake transitions compared with Hcrt stimulation alone. We also built a conductance-based computational model of Hcrt-LC circuitry that recapitulates our behavioral results using LC neurons as the main effectors of Hcrt signaling. These results establish the Hcrt-LC connection as a critical integrator-effector circuit that regulates NREM sleep/wake behavior during the inactive period. This coupling of distinct neuronal systems can be generalized to other hypothalamic integrator nuclei with downstream effector/output populations in the brain.ChR2 | norepinephrine | step function opsin T he neural basis of wakefulness and arousal is thought to depend on subcortical populations of "arousal-promoting" nuclei located in the hypothalamus and brainstem (1, 2). Neurons in the lateral hypothalamus that express hypocretins (Hcrts-also called "orexins"), a pair of neuropeptides produced from the same genetic precursor (3, 4), have been proposed to play a key role in stabilizing wake states by directly projecting throughout the brain to other arousal populations (1,5,6). Electrophysiological recordings of Hcrt neurons show that they are relatively silent during sleep compared with wakefulness, with phasic bursts of activity preceding transitions to wakefulness (7,8). Loss-of-function perturbation of the Hcrts or their receptors causes a narcolepsy phenotype (9-11). When centrally administered, the Hcrts increase the time spent awake and decrease nonrapid eye movement (NREM) and rapid eye movement (REM) sleep (12, 13).Although it is evident that Hcrts promote wakefulness and arousal, it is unknown which downstream structures are necessary and/or sufficient to mediate their effects. Hcrt neurons project diffusely throughout the brain (14), and it is possible that their effects on wakefulness are due to widespread, global postsynaptic targets. Alternatively, Hcrts may affect arousal primarily by projecting to key downstream arousal centers. An intriguing possibility is that Hcrt neurons affect wakefulness by projecting to the locus coeruleus (LC), a noradrenergic structure in the brainstem known to promote wakefulness and arousal (15, 16). Indeed, the LC receives the densest afferent projections from Hcrt neurons (14, 17). Application o...

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Section: Resultsmentioning

confidence: 99%

Mechanism for Hypocretin-mediated sleep-to-wake transitions

Carter

Brill

Bonnavion³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

247

203

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“…Two mouse strains were crossed to label dopaminergic neurons. Homozygous TH::IRES-Cre driver mice (Lindeberg et al, 2004) were crossed with heterozygous Z/EG Cre reporter mice (Novak et al, 2000). Genotypes were determined from mouse-tail DNA samples by PCR for GFP (forward primer: CGA CGT AAA CGG CCA CAA GT; reverse primer: TGT TGT AGT TGT ACT CCA GC).…”

Section: Methodsmentioning

confidence: 99%

Long-Term Imaging Reveals Dynamic Changes in the Neuronal Composition of the Glomerular Layer

Adam

Mizrahi²

2011

J. Neurosci.

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The mammalian olfactory bulb (OB) contains a rich and highly heterogeneous network of local interneurons (INs). These INs undergo continuous turnover in the adult OB in a process known as "adult neurogenesis." Although the overall magnitude of adult neurogenesis has been estimated, the detailed dynamics of the different subpopulations remains largely unknown. Here we present a novel preparation that enables long-term in vivo time-lapse imaging in the mouse OB through a chronic cranial window in a virtually unlimited number of sessions. Using this preparation, we followed the turnover of a specific neuronal population in the OB, the dopaminergic (DA) neurons, for as long as 9 months. By following the same population over long periods of time, we found clear addition and loss of DA neurons in the glomerular layer. Both cell addition and loss increased over time. The numbers of new DA cells were consistently and significantly higher than lost DA cells, suggesting a net increase in the size of this particular population with age. Over a 9 month period of adult life, the net addition of DA neurons reached ϳ13%. Our data argue that the fine composition of the bulbar IN network changes throughout adulthood rather than simply being replenished.

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“…1 A, available at www.jneurosci.org as supplemental material). To spatially restrict ablation of Psmc1, Psmc1 fl/fl mice were crossed with Cre deletor mouse strains, expressing Cre recombinase under the control of either the CaMKII␣ promoter (Psmc1 fl/fl ;CaMKII␣-Cre) or from the TH locus (Psmc1 fl/fl ;TH Cre ) (Lindeberg et al, 2002(Lindeberg et al, , 2004. The CaMKII␣ promoter directs expression to neurons predominantly in forebrain regions (Burgin et al, 1990;Mayford et al, 1996;Tsien et al, 1996).…”

Section: Generation Of Neuron-specific 26s Proteasome-depleted Micementioning

confidence: 99%

“…Therefore, we generated a reproducible conditional knock-out mouse using the Cre/loxP method for an essential subunit of the 19S RP, PSMC1 (Rpt2/S4). Two Cre deletor mouse strains were used to spatially restrict inactivation of Psmc1 to predominantly the forebrain or substantia nigra, expressing Cre recombinase either under the control of the calcium calmodulin-dependent protein kinase II␣ (CaMKII␣) promoter or from the tyrosine hydroxylase (TH ) locus (Lindeberg et al, 2002(Lindeberg et al, , 2004, respectively. We show here that loss of PSMC1 leads to 26S proteasome depletion, which causes neurodegeneration and the formation of intraneuronal Lewylike inclusions resembling human pale bodies (PBs) in neurons of the nigrostriatal pathway and forebrain.…”

Section: Introductionmentioning

confidence: 99%

Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies

Bedford

Hay²,

Devoy³

et al. 2008

J. Neurosci.

291

254

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Ubiquitin-positive intraneuronal inclusions are a consistent feature of the major human neurodegenerative diseases, suggesting that dysfunction of the ubiquitin proteasome system is central to disease etiology. Research using inhibitors of the 20S proteasome to model Parkinson's disease is controversial. We report for the first time that specifically 26S proteasomal dysfunction is sufficient to trigger neurodegenerative disease. Here, we describe novel conditional genetic mouse models using the Cre/loxP system to spatially restrict inactivation of Psmc1 (Rpt2/S4) to neurons of either the substantia nigra or forebrain (e.g., cortex, hippocampus, and striatum). PSMC1 is an essential subunit of the 26S proteasome and Psmc1 conditional knock-out mice display 26S proteasome depletion in targeted neurons, in which the 20S proteasome is not affected. Impairment of specifically ubiquitin-mediated protein degradation caused intraneuronal Lewy-like inclusions and extensive neurodegeneration in the nigrostriatal pathway and forebrain regions. Ubiquitin and ␣-synuclein neuropathology was evident, similar to human Lewy bodies, but interestingly, inclusion bodies contained mitochondria. We support this observation by demonstrating mitochondria in an early form of Lewy body (pale body) from Parkinson's disease patients. The results directly confirm that 26S dysfunction in neurons is involved in the pathology of neurodegenerative disease. The model demonstrates that 26S proteasomes are necessary for normal neuronal homeostasis and that 20S proteasome activity is insufficient for neuronal survival. Finally, we are providing the first reproducible genetic platform for identifying new therapeutic targets to slow or prevent neurodegeneration.

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Transgenic expression of Cre recombinase from the tyrosine hydroxylase locus

Cited by 197 publications

References 32 publications

Mechanism for Hypocretin-mediated sleep-to-wake transitions

Mechanism for Hypocretin-mediated sleep-to-wake transitions

Long-Term Imaging Reveals Dynamic Changes in the Neuronal Composition of the Glomerular Layer

Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies

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