DARPP-32 genomic fragments drive Cre expression in postnatal striatum

Bogush, Alexey; McCarthy, Lois E.; Tian, Chai; Olm, Vicki; Gieringer, Tracy; Ivković, Sanja; Ehrlich, Michelle E.

doi:10.1002/gene.20118

Cited by 27 publications

(50 citation statements)

References 33 publications

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“…We speculated that the increased nocturnal activity of RIIβ KO mice was caused by RIIβ deficiency in the striatum (5). To examine this hypothesis we generated mice with selective RIIβ reexpression in striatal medium spiny neurons (MSNs) by using Darpp32-Cre (D32-Cre) mice (20). In D32-Cre/ RΙΙβ lox/− mice, RΙΙβ was exclusively reexpressed in the striatum and remained undetectable in any of the peripheral tissues examined, including WAT, BAT, pituitary, thyroid, and adrenal gland.…”

Section: Specific Riiβ Reexpression In Striatum Reverses the Hyperactmentioning

confidence: 99%

Deficiency of the RIIβ subunit of PKA affects locomotor activity and energy homeostasis in distinct neuronal populations

Zheng¹,

Yang²,

Sikorski³

et al. 2013

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

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Targeted disruption of RIIβ-protein kinase A (PKA) in mice leads to a lean phenotype, increased nocturnal locomotor activity, and activation of brown adipose tissue. Because RIIβ is abundantly expressed in both white and brown adipose tissue as well as the brain, the contribution of neuronal vs. peripheral PKA to these phenotypes was investigated. We used a Cre-Lox strategy to reexpress RIIβ in a tissue-specific manner in either adipocytes or neurons. Mice with adipocyte-specific RIIβ reexpression remained hyperactive and lean, but pan-neuronal RIIβ reexpression reversed both phenotypes. Selective RIIβ reexpression in all striatal medium spiny neurons with Darpp32-Cre corrected the hyperlocomotor phenotype, but the mice remained lean. Further analysis revealed that RIIβ reexpression in D2 dopamine receptor-expressing medium spiny neurons corrected the hyperlocomotor phenotype, which demonstrated that the lean phenotype in RIIβ-PKA-deficient mice does not develop because of increased locomotor activity. To identify the neurons responsible for the lean phenotype, we used specific Cre-driver mice to reexpress RIIβ in agouti-related peptide (AgRP)-, proopiomelanocortin (POMC)-, single-minded 1 (Sim1)-, or steroidogenic factor 1 (SF1)-expressing neurons in the hypothalamus, but observed no rescue of the lean phenotype. However, when RIIβ was reexpressed in multiple regions of the hypothalamus and striatum driven by Rip2-Cre, or specifically in GABAergic neurons driven by Vgat-ires-Cre, both the hyperactive and lean phenotypes were completely corrected. Bilateral injection of adeno-associated virus1 (AAV1)-Cre directly into the hypothalamus caused reexpression of RIIβ and partially reversed the lean phenotype. These data demonstrate that RIIβ-PKA deficiency in a subset of hypothalamic GABAergic neurons leads to the lean phenotype.mouse genetics | obesity | exercise | cAMP W e reported previously that mice lacking the protein kinase A (PKA) regulatory subunit RΙΙβ (RΙΙβ KO) exhibit a 50% reduction in white adipose tissue (WAT) and are resistant to dietinduced obesity (DIO) and diabetes (1, 2). These mice have an extended lifespan and show diminished age-related metabolic dysfunctions such as fatty liver and insulin resistance (3). Compared with their WT littermates, RIIβ KO mice have normal to slightly increased food intake, a twofold increase in nocturnal physical activity, and a basal metabolic rate (VO 2 consumption) that is higher than WT if calculated based on total body weight (4-6). RIIβ is highly expressed in the mouse CNS, brown adipose tissue (BAT), and WAT with limited expression elsewhere (1, 7-9). At the molecular level, RIIβ deficiency is accompanied by a compensatory increase in the PKA regulatory subunit RIα, which results in increased basal PKA activity and decreased total C subunit activity in brain, BAT, and WAT (1). These changes in PKA subunit composition may also alter PKA holoenzyme localization to specific signaling complexes because RIIβ has a much higher affinity for anchoring proteins (AKAPs)...

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Section: Specific Riiβ Reexpression In Striatum Reverses the Hyperactmentioning

confidence: 99%

Deficiency of the RIIβ subunit of PKA affects locomotor activity and energy homeostasis in distinct neuronal populations

Zheng¹,

Yang²,

Sikorski³

et al. 2013

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

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“…The authors concluded that MSN polyQ-htt expression is not sufficient to cause disease, but in fact, cell-autonomous abnormalities of the MSNs were demonstrated. 6) As previously noted, we used DARPP-32 genomic fragments (also known as D9) to direct expression to the MSNs and exclude other forebrain neurons [43]. These mice (D9-N171-98Q) develop MSN inclusions, progressive motor abnormalities, failure to gain weight after 12 months of age, striatal transcriptional abnormalities, decreased striatal mitochondrial complex II activity, and in aged animals, resistance to quinolinic acid-induced excitotoxicity [5,44,45], discussed in detail as follows.…”

Section: Evidence Supporting Intrinsic Vulnerability Of Msns In Hdmentioning

confidence: 99%

“…This latter study used Emx1-Cre to conditionally delete BDNF in the cortex, and recombination occurs as early as embryonic day 11, long before BDNF is down-regulated in HD. Regulation of transcription by BDNF requires signal transduction systems altered in HD (e.g., phosphatidylinositol-3-kinase and cdk5) [43,178]. Increased BDNF, either by intraventricular injection or prenatal transgenic overexpression, ameliorates the severity of the phenotype in R6 and YAC128 mice, whereas crossing with BDNF heterozygote-null mice exacerbates the phenotype [62,63,179].…”

Section: Bdnf Deficitmentioning

confidence: 99%

Huntington's Disease and the Striatal Medium Spiny Neuron: Cell-Autonomous and Non-Cell-Autonomous Mechanisms of Disease

Ehrlich

2012

Neurotherapeutics

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126

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Huntington's disease is an autosomal dominant disorder caused by a mutation in the gene encoding the protein huntingtin on chromosome 4. The mutation is an expanded CAG repeat in the first exon, encoding a polyglutamine tract. If the polyglutamine tract is >40, penetrance is 100% and death is inevitable. Despite the widespread expression of huntingtin, HD has long been considered primarily as a disease of the striatum. It is characterized by selective vulnerability with dysfunction followed by death of the medium size spiny neuron. Considerable effort is being expended to determine whether striatal damage is cell-autonomous, non-cell-autonomous, requiring cell-cell and region to region communication, or both. We review data supporting both mechanisms. We also attempt to organize the data into common mechanisms that may arise outside the medium, spiny neuron, but ultimately have their greatest impact in the striatum. characterized by selective, or perhaps better described as differential [2], vulnerability with degeneration and death of the medium spiny neuron (MSN). The Gamma-aminobutyric acid (GABA)ergic MSN represents 95% of striatal neurons. They are all output neurons, with extensive intra-striatal connections with other MSNs and striatal interneurons. For the last two decades, increased attention has been paid to the pathology in the cortex (particularly in layers V and VI [3]), which contains the corticostriatal projection neurons.Prior to identification of the HTT gene, the huntingtin protein, and the nature of its mutation, it was assumed that selective neuronal vulnerability in HD would be conferred by the expression pattern of the mutated protein (polyQ-htt). As it is now apparent, htt is expressed throughout the nervous system and periphery, without a preference for, or higher level in, MSNs or cortical projection neurons [4].In the absence of enriched expression of a mutated protein, neuronal subtype vulnerability was assumed to arise from unique protein interactions. For example, in the study of another polyQ disease, spinocerebellar ataxia 7, the interaction between ataxin-7 and the homeobox protein CRX in the retina was reported to specifically lead to pathology [5]. In a followup study, the specific role of CRX was not confirmed [6]. To complicate matters further, the same group recently demonstrated that the interactions between a mutant polyQ protein, Ataxin-1, and 14-3-3epsilon differentially affects disease state in cerebellum and brainstem, both of which are vulnerable regions [7]. The regional distribution of the described huntingtin interacting proteins by and large does not explain MSN vulnerability [8][9][10]. One exception is the htt interacting protein RASD2/Rhes (Ras homologue enriched in striatum), which is striatal-specific. It is a small G-protein that mediates sumoylation of polyQ-htt, and thereby its neurotoxicity.

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“…We demonstrated that the promoter and intragenic regions of DARPP-32, collectively termed D9 (genomic elements from DARPP-32 encompassing 9 kb), selectively direct transgene expression to MSNs (Bogush et al, 2005;Brown et al, 2008), whereas the 2.1 kb of 5Ј UTR sequence alone does not direct expression to any region of the CNS (Blau et al, 1995). In this study, we sought to characterize the function of a region in intron IV of the Ppp1r1b gene, also known as H10.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we sought to characterize the function of a region in intron IV of the Ppp1r1b gene, also known as H10. Our interest in this particular sequence arose from its high conservation among species and its location in a region of the DARPP-32 gene that is included in D9 (Bogush et al, 2005). After the identification of multiunit complexes between H10 and striatal nuclear proteins, the protein-binding site was determined.…”

Section: Introductionmentioning

confidence: 99%

Egr-1 Induces DARPP-32 Expression in Striatal Medium Spiny Neurons via a Conserved Intragenic Element

et al. 2012

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DARPP-32 (dopamine and adenosine 3Ј, 5Ј-cyclic monophosphate cAMP-regulated phosphoprotein, 32 kDa) is a striatal-enriched protein that mediates signaling by dopamine and other first messengers in the medium spiny neurons. The transcriptional mechanisms that regulate striatal DARPP-32 expression remain enigmatic and are a subject of much interest in the efforts to induce a striatal phenotype in stem cells. We report the identification and characterization of a conserved region, also known as H10, in intron IV of the gene that codes for DARPP-32 (Ppp1r1b). This DNA sequence forms multiunit complexes with nuclear proteins from adult and embryonic striata of mice and rats. Purification of proteins from these complexes identified early growth response-1 (Egr-1). The interaction between Egr-1 and H10 was confirmed in vitro and in vivo by super-shift and chromatin immunoprecipitation assays, respectively. Importantly, brain-derived neurotrophic factor (BDNF), a known inducer of DARPP-32 and Egr-1 expression, enhanced Egr-1 binding to H10 in vitro. Moreover, overexpression of Egr-1 in primary striatal neurons induced the expression of DARPP-32, whereas a dominantnegative Egr-1 blocked DARPP-32 induction by BDNF. Together, this study identifies Egr-1 as a transcriptional activator of the Ppp1r1b gene and provides insight into the molecular mechanisms that regulate medium spiny neuron maturation.

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DARPP-32 genomic fragments drive Cre expression in postnatal striatum

Cited by 27 publications

References 33 publications

Deficiency of the RIIβ subunit of PKA affects locomotor activity and energy homeostasis in distinct neuronal populations

Deficiency of the RIIβ subunit of PKA affects locomotor activity and energy homeostasis in distinct neuronal populations

Huntington's Disease and the Striatal Medium Spiny Neuron: Cell-Autonomous and Non-Cell-Autonomous Mechanisms of Disease

Egr-1 Induces DARPP-32 Expression in Striatal Medium Spiny Neurons via a Conserved Intragenic Element

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