Molecular basis of dendritic atrophy and activity in stress susceptibility

Francis, T. Chase; Chandra, Ramesh; Gaynor, A. Smith; Konkalmatt, Prasad; Metzbower, Sarah R.; Evans, Brianna; Engeln, Michel; Blanpied, Thomas A.; Lobo, Mary Kay

doi:10.1038/mp.2017.178

Cited by 81 publications

(114 citation statements)

References 50 publications

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“…The present study was designed to address this deficiency in the field.Our two cell types of choice are both principal GABAergic MSNs of the NAc, a forebrain region implicated in reward and motivation. Both MSN subtypes respond to dopamine, but do so through the activity of different dopamine receptors 17,18 , display different physiology in response to reward-related stimuli 19-21 , and generate different behavioral outcomes [22][23][24][25][26][27][28][29] . Whole cell-FACS 17 and TRAP 12 have been used to distinguish between D1-and D2-MSNs, but the two methods have not been compared directly and these prior studies focused on the entire striatal complex of which the NAc represents a small sub-region.…”

mentioning

confidence: 99%

Biology and Bias in Cell Type-Specific RNAseq of Nucleus Accumbens Medium Spiny Neurons

Kronman

Richter

Labonté

et al. 2018

Preprint

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Isolation of cell populations is untangling complex biological interactions, but studies comparing methodologies lack in vivo complexity and draw limited conclusions about the types of transcripts identified by each technique. Furthermore, few studies compare FACS-based techniques to ribosomal affinity purification, and none do so genome-wide. We addressed this gap by systematically comparing nuclear-FACS, whole cell-FACS, and RiboTag affinity purification in the context of D1 or D2 dopamine receptor-expressing medium spiny neuron (MSN) subtypes of the nucleus accumbens (NAc), a key brain reward region. We find that nuclear-FACS-seq generates a substantially longer list of differentially expressed genes between these cell types, and a significantly larger number of neuropsychiatric GWAS hits than the other two methods. RiboTag-seq has much lower coverage of the transcriptome than the other methods, but very efficiently distinguishes D1-and D2-MSNs. We also demonstrate differences between D1-and D2-MSNs with respect to RNA localization, suggesting fundamental cell type differences in mechanisms of transcriptional regulation and subcellular transport of RNAs. Together, these findings guide the field in selecting the RNAseq method that best suits the scientific questions under investigation.ignore the important question about which RNA species are isolated by the latter methods that capture active translation [12][13][14] . Those studies that include such a comparison have relatively simple biological endpoints like method repeatability and contamination, and lack the comprehensiveness of a whole-genome analysis 10,15,16 .The absence of a methodical comparison of whole cell-FACS, nuclear-FACS, and RiboTag affinity purification is becoming increasingly problematic as an increasing number of studies using these techniques are published and their results taken at face value. These techniques capture different cellular processes while simultaneously defining the same cellular identity. Only a head-to-head comparison for the same cell types can demonstrate if they predominantly capture differences or similarities, and identify the nature of those biases. The present study was designed to address this deficiency in the field.Our two cell types of choice are both principal GABAergic MSNs of the NAc, a forebrain region implicated in reward and motivation. Both MSN subtypes respond to dopamine, but do so through the activity of different dopamine receptors 17,18 , display different physiology in response to reward-related stimuli 19-21 , and generate different behavioral outcomes [22][23][24][25][26][27][28][29] . Whole cell-FACS 17 and TRAP 12 have been used to distinguish between D1-and D2-MSNs, but the two methods have not been compared directly and these prior studies focused on the entire striatal complex of which the NAc represents a small sub-region. Here, we use all three RNA isolation methods -whole cell-FACS, nuclear-FACS, and RiboTag -to provide a deeper characterization of these behaviorally relevant NAc cell typ...

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mentioning

confidence: 99%

Biology and Bias in Cell Type-Specific RNAseq of Nucleus Accumbens Medium Spiny Neurons

Kronman

Richter

Labonté

et al. 2018

Preprint

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“…Factors, such as route of cocaine administration, methods of extraction, and potential co-morbid mental disorders in human subjects, such as major depression or bipolar disorder (See Table S1 in 1 ). These differences may reflect neuropsychopathological differences in genotype and phenotype regulation patterns in the NAc and in MSN subtypes linked to broad forms of neuropathology [9][10][11][47][48][49] . Additionally, the analysis in bulk tissue could mask cell subtype specific effects that we have observed in rodent tissue.…”

Section: Resultsmentioning

confidence: 99%

Cocaine-induced neuron subtype mitochondrial dynamics through Egr3 transcriptional regulation

Cole

Chandra

Harris

et al. 2020

Preprint

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AbstractMitochondrial function is required for brain energy homeostasis and neuroadaptation. Recent studies demonstrated that cocaine affects mitochondrial dynamics within the nucleus accumbens (NAc) and mitochondria are differentially regulated by cocaine in dopamine receptor-1 (D1) containing medium spiny neurons (MSNs) vs dopamine receptor-2 (D2)-MSNs. Here, it is demonstrated that cocaine enhances binding of the transcription factor, early growth response factor 3 (Egr3), to nuclear genes involved in mitochondrial function and dynamics. Further, cocaine exposure regulates mRNA of these mitochondria-associated nuclear genes in both contingent or noncontingent cocaine administration and in both rodent models and human postmortem tissue. Interestingly, several mitochondrial genes showed distinct profiles of expression in D1-MSNs vs D2-MSNs, with cocaine exposure generally increasing mitochondrial-associated nuclear gene expression in D1-MSNs vs suppression in D2-MSNs. Subsequent experiments demonstrated that Egr3 overexpression in D1-MSNs enhances the expression of several mitochondrial-associated nuclear genes. By contrast, blunting Egr3 expression blocks cocaine enhancement of the mitochondrial-associated transcriptional coactivator, peroxisome proliferator-activated receptor gamma coactivator (PGC1α), and the mitochondrial fission molecule, dynamin related protein 1 (Drp1). Finally, reducing Egr3 expression attenuates the cocaine-induced enhancement of small-sized mitochondria, demonstrating that Egr3 regulates mitochondrial morphological adaptations. Collectively, these studies demonstrate cocaine exposure impacts Egr3 transcriptional regulation of mitochondria-related nuclear gene transcripts and mitochondrial dynamics, with implications for mechanisms underlying neuronal function and plasticity occurring with cocaine exposure.

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“…Additionally, reduced excitatory input can induce changes in intrinsic excitability at the postsynaptic level to maintain homeostasis in circuit activity 70 . Our group has recently described this mechanism for stress-induced dendritic atrophy in ventral striatal D1-MSNs 36,38,71 . Further, our previous studies demonstrate reduced inhibition of ventral striatal D1-MSNs in D1-Cre-flTrkB mice 72 .…”

Section: Discussionmentioning

confidence: 99%

Individual differences in stereotypy and neuron subtype translatome with TrkB deletion

Engeln

Song

Chandra

et al. 2019

Preprint

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AbstractMotor stereotypies occurring in early-onset neuropsychiatric diseases are associated with dysregulated basal ganglia direct-pathway activity. Disruptions in network connectivity through impaired neuronal structure have been implicated in both rodents and humans. However, the neurobiological mechanisms leading to direct-pathway neuron disconnectivity in stereotypy remain poorly understood. We have a mouse line with Tropomyosin receptor kinase B (TrkB) receptor deletion from D1-expressing cells (D1-Cre-flTrkB) in which a subset of animals shows repetitive rotations and head tics with juvenile onset. Here we demonstrate these behaviors may be associated with abnormal direct-pathway activity by reducing rotations using chemogenetic inhibition of dorsal striatum D1-medium spiny neurons (D1-MSNs) in both juvenile and young adult mice. Taking advantage of phenotypical differences in animals with similar genotype, we then interrogated the D1-MSN specific translatome associated with repetitive behavior by using RNA-sequencing of ribosome-associated mRNA. Detailed translatome analysis followed by multiplexed gene expression assessment revealed profound alterations in neuronal projection and synaptic structure related genes in stereotypy mice. Examination of neuronal morphology demonstrated dendritic atrophy and dendritic spine loss in dorsal striatum D1-MSNs from mice with repetitive behavior. Together, our results uncover phenotype-specific molecular alterations in D1-MSNs that relate to morphological adaptations in mice displaying stereotypy behavior.

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Molecular basis of dendritic atrophy and activity in stress susceptibility

Cited by 81 publications

References 50 publications

Biology and Bias in Cell Type-Specific RNAseq of Nucleus Accumbens Medium Spiny Neurons

Biology and Bias in Cell Type-Specific RNAseq of Nucleus Accumbens Medium Spiny Neurons

Cocaine-induced neuron subtype mitochondrial dynamics through Egr3 transcriptional regulation

Individual differences in stereotypy and neuron subtype translatome with TrkB deletion

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