Transgenic Rat Project – Publications

@article{Strong:2020tx,

title = {Chemogenetic selective manipulation of nucleus accumbens medium spiny neurons bidirectionally controls alcohol intake in male and female rats},

author = {C. E. Strong and D. P. Hagarty and A. Brea Guerrero and K. J. Schoepfer and S. M. Cajuste and M. Kabbaj},

url = {https://pubmed.ncbi.nlm.nih.gov/33154463/},

doi = {10.1038/s41598-020-76183-2},

isbn = {2045-2322},

year  = {2020},

date = {2020-11-05},

urldate = {2020-11-05},

journal = {Scientific Reports},

volume = {10},

number = {1},

pages = {19178},

abstract = {The nucleus accumbens (NAc), considered the hub of reward circuitry, is comprised of two medium spiny neuron (MSN) subtypes that are classified by their enrichment of dopamine 1 (D1) or 2 (D2) receptors. While reports indicate that alcohol increases excitatory neurotransmission exclusively on NAc D1-MSNs in male rats, it remains unknown how NAc MSNs control alcohol intake in either sex. Therefore, this study investigated how NAc MSNs mediate alcohol intake by using Drd1a-iCre and Drd2-iCre transgenic rats of both sexes. Intra-NAc infusions of Cre-inducible viral vectors containing stimulatory (hM3Dq) or inhibitory (hM4Di) designer receptors exclusively activated by designer drugs (DREADDs) were delivered after 4-weeks of alcohol intake, and clozapine-N-oxide (CNO) was administered to selectively manipulate NAc MSNs. Our results show that activation of NAc D1-MSNs increased alcohol intake 1-, 4-, and 24-h after the start of drinking while inhibition decreased it 1-h after the start of drinking, with no sex differences observed at any time point. Activation of NAc D2-MSNs had no impact on alcohol intake while inhibition increased alcohol intake in Drd2-iCre rats for 1-h in males and 4-h in females. These findings suggest opposing roles for how NAc D1- and D2-MSNs modulate alcohol intake in rats of both sexes.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{10.7554/eLife.59716,

title = {An approach for long-term, multi-probe Neuropixels recordings in unrestrained rats},

author = {Thomas Zhihao Luo and Adrian Gopnik Bondy and Diksha Gupta and Verity Alexander Elliott and Charles D Kopec and Carlos D Brody},

editor = {Lisa Giocomo and Laura L Colgin and Lisa Giocomo and Anne K Churchland},

url = {https://pubmed.ncbi.nlm.nih.gov/33089778/},

doi = {10.7554/eLife.59716},

issn = {2050-084X},

year  = {2020},

date = {2020-10-01},

journal = {eLife},

volume = {9},

pages = {e59716},

publisher = {eLife Sciences Publications, Ltd},

abstract = {The use of Neuropixels probes for chronic neural recordings is in its infancy and initial studies leave questions about long-term stability and probe reusability unaddressed. Here, we demonstrate a new approach for chronic Neuropixels recordings over a period of months in freely moving rats. Our approach allows multiple probes per rat and multiple cycles of probe reuse. We found that hundreds of units could be recorded for multiple months, but that yields depended systematically on anatomical position. Explanted probes displayed a small increase in noise compared to unimplanted probes, but this was insufficient to impair future single-unit recordings. We conclude that cost-effective, multi-region, and multi-probe Neuropixels recordings can be carried out with high yields over multiple months in rats or other similarly sized animals. Our methods and observations may facilitate the standardization of chronic recording from Neuropixels probes in freely moving animals.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Baez-Cordero:2020aa,

title = {Unbalanced Inhibitory/Excitatory Responses in the Substantia Nigra Pars Reticulata Underlie Cannabinoid-Related Slowness of Movements},

author = {Ana S Báez-Cordero and Ana K Pimentel-Farfan and Teresa Pe{~n}a-Rangel and Pavel E Rueda-Orozco},

url = {http://www.jneurosci.org/content/40/30/5769.abstract},

doi = {10.1523/JNEUROSCI.0045-20.2020},

year  = {2020},

date = {2020-07-22},

journal = {The Journal of Neuroscience},

volume = {40},

number = {30},

pages = {5769},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{10.7554/eLife.57313,

title = {Different theta frameworks coexist in the rat hippocampus and are coordinated during memory-guided and novelty tasks},

author = {Victor J López-Madrona and Elena Pérez-Montoyo and Efrén Álvarez-Salvado and David Moratal and Oscar Herreras and Ernesto Pereda and Claudio R Mirasso and Santiago Canals},

editor = {Martin Vinck and Laura L Colgin and Antonio Fernandez-Ruiz},

url = {https://pubmed.ncbi.nlm.nih.gov/32687054/},

doi = {10.7554/eLife.57313},

issn = {2050-084X},

year  = {2020},

date = {2020-07-01},

journal = {eLife},

volume = {9},

pages = {e57313},

publisher = {eLife Sciences Publications, Ltd},

abstract = {Hippocampal firing is organized in theta sequences controlled by internal memory processes and by external sensory cues, but how these computations are coordinated is not fully understood. Although theta activity is commonly studied as a unique coherent oscillation, it is the result of complex interactions between different rhythm generators. Here, by separating hippocampal theta activity in three different current generators, we found epochs with variable theta frequency and phase coupling, suggesting flexible interactions between theta generators. We found that epochs of highly synchronized theta rhythmicity preferentially occurred during behavioral tasks requiring coordination between internal memory representations and incoming sensory information. In addition, we found that gamma oscillations were associated with specific theta generators and the strength of theta-gamma coupling predicted the synchronization between theta generators. We propose a mechanism for segregating or integrating hippocampal computations based on the flexible coordination of different theta frameworks to accommodate the cognitive needs.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Garcia-Keller8463,

title = {Relapse-Associated Transient Synaptic Potentiation Requires Integrin-Mediated Activation of Focal Adhesion Kinase and Cofilin in D1-Expressing Neurons},

author = {Constanza Garcia-Keller and Michael D Scofield and Daniela Neuhofer and Swathi Varanasi and Matthew T Reeves and Brandon Hughes and Ethan Anderson and Christopher T Richie and Carlos Mejias-Aponte and James Pickel and Bruce T Hope and Brandon K Harvey and Christopher W Cowan and Peter W Kalivas},

url = {https://pubmed.ncbi.nlm.nih.gov/33051346/},

doi = {10.1523/JNEUROSCI.2666-19.2020},

issn = {0270-6474},

year  = {2020},

date = {2020-01-01},

urldate = {2020-01-01},

journal = {Journal of Neuroscience},

volume = {40},

number = {44},

pages = {8463--8477},

publisher = {Society for Neuroscience},

abstract = {Relapse to drug use can be initiated by drug-associated cues. The intensity of cue-induced drug seeking in rodent models correlates with the induction of transient synaptic potentiation (t-SP) at glutamatergic synapses in the nucleus accumbens core (NAcore). Matrix metalloproteinases (MMPs) are inducible endopeptidases that degrade extracellular matrix (ECM) proteins, and reveal tripeptide Arginine-Glycine-Aspartate (RGD) domains that bind and signal through integrins. Integrins are heterodimeric receptors composed of αβ subunits, and a primary signaling kinase is focal adhesion kinase (FAK). We previously showed that MMP activation is necessary for and potentiates cued reinstatement of cocaine seeking, and MMP-induced catalysis stimulates β3-integrins to induce t-SP. Here, we determined whether β3-integrin signaling through FAK and cofilin (actin depolymerization factor) is necessary to promote synaptic growth during t-SP. Using a small molecule inhibitor to prevent FAK activation, we blocked cued-induced cocaine reinstatement and increased spine head diameter (dh). Immunohistochemistry on NAcore labeled spines with ChR2-EYFP virus, showed increased immunoreactivity of phosphorylation of FAK (p-FAK) and p-cofilin in dendrites of reinstated animals compared with extinguished and yoked saline, and the p-FAK and cofilin depended on β3-integrin signaling. Next, male and female transgenic rats were used to selectively label D1 or D2 neurons with ChR2-mCherry. We found that p-FAK was increased during drug seeking in both D1 and D2-medium spiny neurons (MSNs), but increased p-cofilin was observed only in D1-MSNs. These data indicate that β3-integrin, FAK and cofilin constitute a signaling pathway downstream of MMP activation that is involved in promoting the transient synaptic enlargement in D1-MSNs induced during reinstated cocaine by drug-paired cues.SIGNIFICANCE STATEMENT Drug-associated cues precipitate relapse, which is correlated with transient synaptic enlargement in the accumbens core. We showed that cocaine cue-induced synaptic enlargement depends on matrix metalloprotease signaling in the extracellular matrix (ECM) through β3-integrin to activate focal adhesion kinase (FAK) and phosphorylate the actin binding protein cofilin. The nucleus accumbens core (NAcore) contains two predominate neuronal subtypes selectively expressing either D1-dopamine or D2-dopamine receptors. We used transgenic rats to study each cell type and found that cue-induced signaling through cofilin phosphorylation occurred only in D1-expressing neurons. Thus, cocaine-paired cues initiate cocaine reinstatement and synaptic enlargement through a signaling cascade selectively in D1-expressing neurons requiring ECM stimulation of β3-integrin-mediated phosphorylation of FAK (p-FAK) and cofilin.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Prasad880,

title = {Complementary Roles for Ventral Pallidum Cell Types and Their Projections in Relapse},

author = {Asheeta A Prasad and Caroline Xie and Chanchanok Chaichim and Jennifer H Nguyen and Hannah E McClusky and Simon Killcross and John M Power and Gavan P McNally},

url = {https://pubmed.ncbi.nlm.nih.gov/31818977/},

doi = {10.1523/JNEUROSCI.0262-19.2019},

issn = {0270-6474},

year  = {2020},

date = {2020-01-01},

journal = {Journal of Neuroscience},

volume = {40},

number = {4},

pages = {880--893},

publisher = {Society for Neuroscience},

abstract = {The ventral pallidum (VP) is a key node in the neural circuits controlling relapse to drug seeking. How this role relates to different VP cell types and their projections is poorly understood. Using male rats, we show how different forms of relapse to alcohol-seeking are assembled from VP cell types and their projections to lateral hypothalamus (LH) and ventral tegmental area (VTA). Using RNAScope in situ hybridization to characterize activity of different VP cell types during relapse to alcohol-seeking provoked by renewal (context-induced reinstatement), we found that VP Gad1 and parvalbumin (PV), but not vGlut2, neurons show relapse-associated changes in c-Fos expression. Next, we used retrograde tracing, chemogenetic, and electrophysiological approaches to study the roles of VPGad1 and VPPV neurons in relapse. We show that VPGad1 neurons contribute to contextual control over relapse (renewal), but not to relapse during reacquisition, via projections to LH, where they converge with ventral striatal inputs onto LHGad1 neurons. This convergence of striatopallidal inputs at the level of individual LHGad1 neurons may be critical to balancing propensity for relapse versus abstinence. In contrast, VPPV neurons contribute to relapse during both renewal and reacquisition via projections to VTA. These findings identify a double dissociation in the roles for different VP cell types and their projections in relapse. VPGad1 neurons control relapse during renewal via projections to LH. VPPV neurons control relapse during both renewal and reacquisition via projections to VTA. Targeting these different pathways may provide tailored interventions for different forms of relapse.SIGNIFICANCE STATEMENT Relapse to drug or reward seeking after a period of extinction or abstinence remains a key impediment to successful treatment. The ventral pallidum, located in the ventral basal ganglia, has long been recognized as an obligatory node in a textquoterightfinal common pathwaytextquoteright for relapse. Yet how this role relates to the considerable VP cellular and circuit heterogeneity is not well understood. We studied the cellular and circuit architecture for VP in relapse control. We show that different forms of relapse have complementary VP cellular and circuit architectures, raising the possibility that targeting these different neural architectures may provide tailored interventions for different forms of relapse.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Back:2019aac,

title = {Neuron-Specific Genome Modification in the Adult Rat Brain Using CRISPR-Cas9 Transgenic Rats.},

author = {Susanne Back and Julie Necarsulmer and Leslie R Whitaker and Lamarque M Coke and Pyry Koivula and Emily J Heathward and Lowella V Fortuno and Yajun Zhang and Grace C Yeh and Heather A Baldwin and Morgan D Spencer and Carlos A Mejias-Aponte and James Pickel and Alexander F Hoffman and Charles E Spivak and Carl R Lupica and Suzanne M Underhill and Susan G Amara and Andrii Domanskyi and Jenni E Anttila and Mikko Airavaara and Bruce T Hope and Kent F Hamra and Christopher T Richie and Brandon K Harvey},

url = {https://www.ncbi.nlm.nih.gov/pubmed/30792150},

doi = {10.1016/j.neuron.2019.01.035},

issn = {1097-4199 (Electronic); 0896-6273 (Linking)},

year  = {2019},

date = {2019-04-03},

urldate = {2019-04-03},

journal = {Neuron},

volume = {102},

number = {1},

pages = {105--119},

address = {Molecular Mechanisms of Cellular Stress and Inflammation Section, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD 21224, USA.},

abstract = {Historically, the rat has been the preferred animal model for behavioral studies. Limitations in genome modification have, however, caused a lag in their use compared to the bevy of available transgenic mice. Here, we have developed several transgenic tools, including viral vectors and transgenic rats, for targeted genome modification in specific adult rat neurons using CRISPR-Cas9 technology. Starting from wild-type rats, knockout of tyrosine hydroxylase was achieved with adeno-associated viral (AAV) vectors expressing Cas9 or guide RNAs (gRNAs). We subsequently created an AAV vector for Cre-dependent gRNA expression as well as three new transgenic rat lines to specifically target CRISPR-Cas9 components to dopaminergic neurons. One rat represents the first knockin rat model made by germline gene targeting in spermatogonial stem cells. The rats described herein serve as a versatile platform for making cell-specific and sequence-specific genome modifications in the adult brain and potentially other Cre-expressing tissues of the rat.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Pardo-Garcia:2019aa,

title = {Ventral Pallidum Is the Primary Target for Accumbens D1 Projections Driving Cocaine Seeking.},

author = {Thibaut R Pardo-Garcia and Constanza Garcia-Keller and Tiffany Penaloza and Christopher T Richie and James Pickel and Bruce T Hope and Brandon K Harvey and Peter W Kalivas and Jasper A Heinsbroek},

doi = {10.1523/JNEUROSCI.2822-18.2018},

issn = {1529-2401 (Electronic); 0270-6474 (Linking)},

year  = {2019},

date = {2019-03-13},

urldate = {2019-03-13},

journal = {J Neurosci},

volume = {39},

number = {11},

pages = {2041--2051},

address = {Department of Neuroscience, University of Michigan, Ann Arbor, Michigan 48109.},

abstract = {Outputs from the nucleus accumbens (NAc) include projections to the ventral pallidum and the ventral tegmental area and subtantia nigra in the ventral mesencephalon. The medium spiny neurons (MSN) that give rise to these pathways are GABAergic and consist of two populations of equal number that are segregated by differentially expressed proteins, including D1- and D2-dopamine receptors. Afferents to the ventral pallidum arise from both D1- and D2-MSNs, whereas the ventral mesencephalon is selectively innervated by D1-MSN. To determine the extent of collateralization of D1-MSN to these axon terminal fields we used retrograde labeling in transgenic mice expressing tdTomato selectively in D1-MSN, and found that a large majority of D1-MSN in either the shell or core subcompartments of the accumbens collateralized to both output structures. Approximately 70% of D1-MSNs projecting to the ventral pallidum collateralized to the ventral mesencephalon, whereas >90% of mesencephalic D1-MSN afferents collateralized to the ventral pallidum. In contrast, <10% of dorsal striatal D1-MSNs collateralized to both the globus pallidus and ventral mesencephalon. D1-MSN activation is required for conditioned cues to induce cocaine seeking. To determine which D1-MSN projection mediates cued cocaine seeking, we selectively transfected D1-MSNs in transgenic rats with an inhibitory Gi-coupled DREADD. Activation of the transfected Gi-DREADD with clozapine-N-oxide administered into the ventral pallidum, but not into the ventral mesencephalon, blocked cue-induced cocaine seeking. These data show that, although accumbens D1-MSNs largely collateralize to both the ventral pallidum and ventral mesencephalon, only D1-MSN innervation of the ventral pallidum is necessary for cue-induced cocaine seeking.SIGNIFICANCE STATEMENT Activity in D1 dopamine receptor-expressing neurons in the NAc is required for rodents to respond to cocaine-conditioned cues and relapse to drug seeking behaviors. The D1-expressing neurons project to both the ventral pallidum and ventral mesencephalon, and we found that a majority of the neurons that innervate the ventral pallidum also collateralize to the ventral mesencephalon. However, despite innervating both structures, only D1 innervation of the ventral pallidum mediates cue-induced cocaine seeking.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{DEMAESTRI2019134,

title = {22 kHz and 55 kHz ultrasonic vocalizations differentially influence neural and behavioral outcomes: Implications for modeling anxiety via auditory stimuli in the rat},

author = {Camila Demaestri and Heather C Brenhouse and Jennifer A Honeycutt},

url = {https://pubmed.ncbi.nlm.nih.gov/30521931/},

doi = {https://doi.org/10.1016/j.bbr.2018.12.005},

issn = {0166-4328},

year  = {2019},

date = {2019-01-01},

journal = {Behavioural Brain Research},

volume = {360},

pages = {134 - 145},

abstract = {The communicative role of ultrasonic vocalizations (USVs) in rats is well established, with distinct USVs indicative of different affective states. USVs in the 22 kHz range are typically emitted by adult rats when in anxiety- or fear-provoking situations (e.g. predator odor, social defeat), while 55 kHz range USVs are typically emitted in appetitive situations (e.g., play, anticipation of reward). Previous work indicates that USVs (real-time and playback) can effectively communicate these affective states and influence changes in behavior and neural activity of the receiver. Changes in cFos activation following 22 kHz USVs have been seen in cortical and limbic regions involved in anxiety, including the basolateral amygdala (BLA). However, it is unclear how USV playback influences cFos activity within the bed nucleus of the stria terminalis (BNST), a region also thought to be critical in processing anxiety-related information, and the nucleus accumbens, a region associated with reward. The present work sought to characterize distinct behavioral, physiological, and neural responses in rats presented with aversive (22 kHz) compared to appetitive (55 kHz) USVs or silence. Our findings show that rats exposed to 22 kHz USVs: 1) engage in anxiety-like behaviors in the elevated zero maze, and 2) show distinct patterns of cFos activation within the BLA and BNST that contrast those seen in 55 kHz playback and silence. Specifically, 22 kHz USVs increased cFos density in the anterodorsal nuclei, while 55 kHz playback increased cFos in the oval nucleus of the BNST, without significant changes within the nucleus accumbens. These results provide important groundwork for leveraging ethologically-relevant stimuli in the rat to improve our understanding of anxiety-related responses in both typical and pathological populations.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Gibson:2018aa,

title = {Distinct Accumbens Shell Output Pathways Promote versus Prevent Relapse to Alcohol Seeking},

author = {Gabrielle D Gibson and Asheeta A Prasad and Philip Jean-Richard-dit-Bressel and Joanna O Y Yau and Zayra E Millan and Yu Liu and Erin J Campbell and Jun Lim and Nathan J Marchant and John M Power and Simon Killcross and Andrew J Lawrence and Gavan P McNally},

url = {https://pubmed.ncbi.nlm.nih.gov/29656870/},

doi = {10.1016/j.neuron.2018.03.033},

isbn = {0896-6273},

year  = {2018},

date = {2018-05-02},

booktitle = {Neuron},

journal = {Neuron},

volume = {98},

number = {3},

pages = {512--520.e6},

publisher = {Elsevier},

abstract = {Contexts exert bi-directional control over relapse to drug seeking. Contexts associated with drug self-administration promote relapse, whereas contexts associated with the absence of self-administration protect against relapse. The nucleus accumbens shell (AcbSh) is a key brain region determining these roles of context. However, the specific cell types, and projections, by which AcbSh serves these dual roles are unknown. Here, we show that contextual control over relapse and abstinence is embedded within distinct output circuits of dopamine 1 receptor (Drd1) expressing AcbSh neurons. We report anatomical and functional segregation of Drd1 AcbSh output pathways during context-induced reinstatement and extinction of alcohol seeking. The AcbSh?ventral tegmental area (VTA) pathway promotes relapse via projections to VTA Gad1 neurons. The AcbSh?lateral hypothalamus (LH) pathway promotes extinction via projections to LH Gad1 neurons. Targeting these opposing AcbSh circuit contributions may reduce propensity to relapse to, and promote abstinence from, drug use.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Zallar:2018aa,

title = {Development and initial characterization of a novel ghrelin receptor CRISPR/Cas9 knockout wistar rat model.},

author = {L J Zallar and B J Tunstall and C T Richie and Y J Zhang and Z B You and E L Gardner and M Heilig and J Pickel and G F Koob and L F Vendruscolo and B K Harvey and L Leggio},

url = {https://www.ncbi.nlm.nih.gov/pubmed/29453460},

doi = {10.1038/s41366-018-0013-5},

issn = {1476-5497 (Electronic); 0307-0565 (Linking)},

year  = {2018},

date = {2018-01-30},

journal = {Int J Obes (Lond)},

address = {Section on Clinical Psychoneuroendocrinology and Neuropsychopharmacology, National Institute on Alcohol Abuse and Alcoholism Division of Intramural Clinical and Biological Research and National Institute on Drug Abuse Intramural Research Program, National Institutes of Health, Bethesda, MD, USA.},

abstract = {BACKGROUND/OBJECTIVES: Ghrelin, a stomach-derived hormone implicated in numerous behaviors including feeding, reward, stress, and addictive behaviors, acts by binding to the growth hormone secretagogue receptor (GHSR). Here, we present the development, verification, and initial characterization of a novel GHSR knockout (KO) Wistar rat model created with CRISPR genome editing. METHODS: Using CRISPR/Cas9, we developed a GHSR KO in a Wistar background. Loss of GHSR mRNA expression was histologically verified using RNAscope in wild-type (WT; n = 2) and KO (n = 2) rats. We tested the effects of intraperitoneal acyl-ghrelin administration on food consumption and plasma growth hormone (GH) concentrations in WT (n = 8) and KO (n = 8) rats. We also analyzed locomotion, food consumption, and body fat composition in these animals. Body weight was monitored from early development to adulthood. RESULTS: The RNAscope analysis revealed an abundance of GHSR mRNA expression in the hypothalamus, midbrain, and hippocampus in WTs, and no observed probe binding in KOs. Ghrelin administration increased plasma GH levels (p = 0.0067) and food consumption (p = 0.0448) in WT rats but not KOs. KO rats consumed less food overall at basal conditions and weighed significantly less compared with WTs throughout development (p = 0.0001). Compared with WTs, KOs presented higher concentrations of brown adipose tissue (BAT; p = 0.0322). CONCLUSIONS: We have verified GHSR deletion in our KO model using histological, physiological, neuroendocrinological, and behavioral measures. Our findings indicate that GHSR deletion in rats is not only associated with a lack of response to ghrelin, but also associated with decreases in daily food consumption and body growth, and increases in BAT. This GHSR KO Wistar rat model provides a novel tool for studying the role of the ghrelin system in obesity and in a wide range of medical and neuropsychiatric disorders.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Hamra:2017aa,

title = {Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production.},

author = {Kent F Hamra and Christopher T Richie and Brandon K Harvey},

url = {https://www.ncbi.nlm.nih.gov/pubmed/28608322},

doi = {10.1007/s11248-017-0025-2},

issn = {1573-9368 (Electronic); 0962-8819 (Linking)},

year  = {2017},

date = {2017-08-01},

urldate = {2017-08-01},

journal = {Transgenic Res},

volume = {26},

number = {4},

pages = {477--489},

address = {Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA. kent.hamra@utsouthwestern.edu.},

abstract = {Long Evans rat strains are applied as research models in a broad spectrum of biomedical fields (>15,800 citations, NCBI PubMed). Here, we report an approach to genetically modify the Long Evans rat germline in donor spermatogonial stem cells. Long Evans rat spermatogonial lines were derived from freshly isolated laminin-binding spermatogonia. Laminin-binding spermatogonia were cultured over multiple passages on fibroblast feeder layers in serum-free culture medium containing GDNF and FGF2. Long Evans rat spermatogonial lines were genetically modified by transposon transduction to express a germline, tdTomato reporter gene. Donor rat spermatogonial lines robustly regenerated spermatogenesis after transplantation into testes of busulfan-treated, allogenic, Long Evans rats. Donor-derived spermatogenesis largely restored testis size in the chemically sterilized, recipient Long Evans rats. Recipient Long Evans rats stably transmitted the tdTomato germline marker to subsequent generations. Overall, Long Evans rat spermatogonial lines provided effective donor germline vectors for genetically modifying Long Evans rats.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Sharpe:2017aab,

title = {Lateral Hypothalamic GABAergic Neurons Encode Reward Predictions that Are Relayed to the Ventral Tegmental Area to Regulate Learning.},

author = {Melissa J Sharpe and Nathan J Marchant and Leslie R Whitaker and Christopher T Richie and Yajun J Zhang and Erin J Campbell and Pyry P Koivula and Julie C Necarsulmer and Carlos Mejias-Aponte and Marisela Morales and James Pickel and Jeffrey C Smith and Yael Niv and Yavin Shaham and Brandon K Harvey and Geoffrey Schoenbaum},

url = {https://www.ncbi.nlm.nih.gov/pubmed/28690111},

doi = {10.1016/j.cub.2017.06.024},

issn = {1879-0445 (Electronic); 0960-9822 (Linking)},

year  = {2017},

date = {2017-07-06},

urldate = {2017-07-06},

journal = {Curr Biol},

volume = {27},

number = {14},

pages = {2089--2100},

address = {National Institute on Drug Abuse, IRP, 251 Bayview Boulevard, Baltimore, MD 21228, USA; Princeton Neuroscience Institute, Princeton University, Washington Road, Princeton, NJ 08544, USA. Electronic address: melissa.sharpe@nih.gov.},

abstract = {Eating is a learned process. Our desires for specific foods arise through experience. Both electrical stimulation and optogenetic studies have shown that increased activity in the lateral hypothalamus (LH) promotes feeding. Current dogma is that these effects reflect a role for LH neurons in the control of the core motivation to feed, and their activity comes under control of forebrain regions to elicit learned food-motivated behaviors. However, these effects could also reflect the storage of associative information about the cues leading to food in LH itself. Here, we present data from several studies that are consistent with a role for LH in learning. In the first experiment, we use a novel GAD-Cre rat to show that optogenetic inhibition of LH gamma-aminobutyric acid (GABA) neurons restricted to cue presentation disrupts the rats' ability to learn that a cue predicts food without affecting subsequent food consumption. In the second experiment, we show that this manipulation also disrupts the ability of a cue to promote food seeking after learning. Finally, we show that inhibition of the terminals of the LH GABA neurons in ventral-tegmental area (VTA) facilitates learning about reward-paired cues. These results suggest that the LH GABA neurons are critical for storing and later disseminating information about reward-predictive cues.},

note = {*First paper describing LE-Tg(GAD1-iCre)3Ottc rat.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Richie:2017aa,

title = {Near-infrared fluorescent protein iRFP713 as a reporter protein for optogenetic vectors, a transgenic Cre-reporter rat, and other neuronal studies.},

author = {Christopher T Richie and Leslie R Whitaker and Keith W Whitaker and Julie Necarsulmer and Heather A Baldwin and Yajun Zhang and Lowella Fortuno and Josh J Hinkle and Pyry Koivula and Mark J Henderson and Wenzhi Sun and Kai Wang and Jeffrey C Smith and Jim Pickel and Na Ji and Bruce T Hope and Brandon K Harvey},

url = {https://www.ncbi.nlm.nih.gov/pubmed/28380331},

doi = {10.1016/j.jneumeth.2017.03.020},

issn = {1872-678X (Electronic); 0165-0270 (Linking)},

year  = {2017},

date = {2017-04-02},

urldate = {2017-04-02},

journal = {J Neurosci Methods},

volume = {284},

pages = {1--14},

address = {Intramural Research Program, National Institute on Drug Abuse, Baltimore, MD 21224, United States.},

abstract = {BACKGROUND: The use of genetically-encoded fluorescent reporters is essential for the identification and observation of cells that express transgenic modulatory proteins. Near-infrared (NIR) fluorescent proteins have superior light penetration through biological tissue, but are not yet widely adopted. NEW METHOD: Using the near-infrared fluorescent protein, iRFP713, improves the imaging resolution in thick tissue sections or the intact brain due to the reduced light-scattering at the longer, NIR wavelengths used to image the protein. Additionally, iRFP713 can be used to identify transgenic cells without photobleaching other fluorescent reporters or affecting opsin function. We have generated a set of adeno-associated vectors in which iRFP713 has been fused to optogenetic channels, and can be expressed constitutively or Cre-dependently. RESULTS: iRFP713 is detectable when expressed in neurons both in vitro and in vivo without exogenously supplied chromophore biliverdin. Neuronally-expressed iRFP713 has similar properties to GFP-like fluorescent proteins, including the ability to be translationally fused to channelrhodopsin or halorhodopsin, however, it shows superior photostability compared to EYFP. Furthermore, electrophysiological recordings from iRFP713-labeled cells compared to cells labeled with mCherry suggest that iRFP713 cells are healthier and therefore more stable and reliable in an ex vivo preparation. Lastly, we have generated a transgenic rat that expresses iRFP713 in a Cre-dependent manner. CONCLUSIONS: Overall, we have demonstrated that iRFP713 can be used as a reporter in neurons without the use of exogenous biliverdin, with minimal impact on viability and function thereby making it feasible to extend the capabilities for imaging genetically-tagged neurons in slices and in vivo.},

note = {*First paper describing LE-Tg (DIO-iRFP)3Ottc rat.

*First paper describing LE-Tg (DIO-iRFP)9Ottc rat.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}