Intact-Brain Analyses Reveal Distinct Information Carried by SNc Dopamine Subcircuits

“Seminal early studies into the information encoded by midbrain DA neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al.”

“We expressed GCaMP6f (Chen et al., 2013) in DMS- or DLS-projecting SNc DA neurons by injecting CAV-cre into the DMS or the DLS, followed by injection of a cre-dependent GCaMP6f construct into the SNc.”

“…neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al., 1996, 2004; Steinberg et al., 2013; Sutton, 1988).”

“…that the DA neurons, which increase their firing in response to these stimuli, may signal salience rather than value (Brischoux et al., 2009; Bromberg-Martin et al., 2010; Matsumoto and Hikosaka, 2009), though controversy on this point remains (Cohen et al., 2012; Fiorillo et al., 2013,…”

“…neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al., 1996, 2004; Steinberg et al., 2013; Sutton, 1988).”

“We began by demonstrating that novel circuit-tracing and -recording methodologies can be effectively combined with whole-brain CLARITY/COLM imaging (Chung et al., 2013; Tomer et al., 2014) to provide a uniquely informative overview of richly defined cell types and their global connectivity motifs…”

“This information notably enriches the ascending spiral model proposed by Haber et al. (2000), which could not make definitive statements about striatal inputs onto DAergic versus non-DAergic midbrain cells.”

“…monitored activity from projection-defined SNc DA neurons during behavior using fiber photometry, a method for collecting population intracellular [Ca2+] fluorescent signals from a genetically encoded Ca2+ indicator such as GCaMP through a single chronic fiber optic implant (Gunaydin et al., 2014).”

“Another study used recordings in awake monkeys to demonstrate a correlation between the depth at which a DA neuron was recorded along the electrode (indicating a more ventromedial location in the midbrain) and the likelihood that it would decrease rather than increase its firing in response to an aversive cue (Matsumo…”

“…target-defined subpopulations are located in different subregions of the VTA, their excitatory synapses are differentially modulated by rewarding and aversive stimuli, and they receive distinct inputs, which can elicit opposite behaviors when selectively recruited (Lammel et al., 2008, 2011, 2012).”

“For example, a recent viral circuit tracing study showed that VTA and SNc DA neurons receive different proportions of inputs from key brain regions (Watabe-Uchida et al., 2012).”

“…functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate in separable circuits.”

“…utilizing a GFP-expressing rabies virus (RVdGGFP) that both lacks the glycoprotein needed to spread transsynaptically and is pseudotyped with EnvA (so that it can infect only neurons expressing TVA, an avian receptor protein normally absent in mammalian cells [Wickersham et al., 2007]).”

“…functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate in separable circuits.”

“Afferents to projection-defined SNc DA neurons also differ; in particular, we here identify new principles of afferent-projection complexity in SNc subcircuits (see Beier et al., 2015 for additional illuminating information regarding VTA subcircuits).”

“…potassium channel antagonist 4-AP (100 mM) to isolate monosynaptic inputs by preventing disynaptic disinhibitory responses through the GABAergic cells of the SNr (with TTX) while enabling ChR2 to drive neurotransmitter release in the absence of action potentials (with 4-AP) (Petreanu et al., 2009).”

“Seminal early studies into the information encoded by midbrain DA neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al.”

“…target-defined subpopulations are located in different subregions of the VTA, their excitatory synapses are differentially modulated by rewarding and aversive stimuli, and they receive distinct inputs, which can elicit opposite behaviors when selectively recruited (Lammel et al., 2008, 2011, 2012).”

“…neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al., 1996, 2004; Steinberg et al., 2013; Sutton, 1988).”

“…neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al., 1996, 2004; Steinberg et al., 2013; Sutton, 1988).”

“…CLARITYmonomer solution containing 1% acrylamide, 0.0125% bis-acrylamide, and 4% PFA and then polymerized at 37 C for 6–7 hr. Brains were passively cleared in SDS Borate Buffer (pH 8.5) at 37 C for 4–5 weeks, equilibrated in Focusclear for imaging and imaged using COLM methods (Tomer et al., 2014).”

“Seminal early studies into the information encoded by midbrain DA neurons suggested that a key function of DA is to transmit reward prediction error signals (Mirenowicz and Schultz, 1996; Schultz et al., 1997; Waelti et al., 2001), a hypothesis concordant with temporal difference learning models (Montague et al.”

“…functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate in separable circuits.”

“However, not all midbrain DA neurons appear to encode similar information in their activity patterns (Bromberg-Martin et al., 2010; Lammel et al., 2014; Roeper, 2013).”

“…leading to the hypothesis that the DA neurons, which increase their firing in response to these stimuli, may signal salience rather than value (Brischoux et al., 2009; Bromberg-Martin et al., 2010; Matsumoto and Hikosaka, 2009), though controversy on this point remains (Cohen et al., 2012;…”

“…which increase their firing in response to these stimuli, may signal salience rather than value (Brischoux et al., 2009; Bromberg-Martin et al., 2010; Matsumoto and Hikosaka, 2009), though controversy on this point remains (Cohen et al., 2012; Fiorillo et al., 2013, 2013; Ungless et al., 2004).”

“…target-defined subpopulations are located in different subregions of the VTA, their excitatory synapses are differentially modulated by rewarding and aversive stimuli, and they receive distinct inputs, which can elicit opposite behaviors when selectively recruited (Lammel et al., 2008, 2011, 2012).”

“Further development of high-throughput processing of these wholebrain datasets will be needed (Chung and Deisseroth, 2013; Kim et al., 2013, 2015) to fully capitalize on this opportunity for the rapid advancement of understanding structure-function relationships in the brain.”

“…CLARITY-optimized light-sheet microscopy (COLM) (Chung et al., 2013; Tomer et al., 2014), along with a variation of rabies-based circuit mapping (Schwarz et al., 2015); the lattermethod (TRIO) operates similarly to previously published rabies-based circuit mapping technologies, utilizing a…”

“d Intact, inclusive approaches to classifying neuronal cell types d Differential brain-wide circuit incorporation of SNc dopamine neuron subpopulations d Opposite valence encoding of shock by projection target- defined SNc neurons d Independently controlled information streams from the SNc to the DMS and DLS Lerner et…”

“…functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate in separable circuits.”

“These regions of the dorsal striatum have been functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate i…”

“Differences in Ih currents are strongly correlated with action potential rebound delays and can influence pacemaking activity (Neuhoff et al., 2002), particularly in the case of calbindin-negative SNc DA neurons (which constitute the majority of the SNc DA neuron population).”

“However, not all midbrain DA neurons appear to encode similar information in their activity patterns (Bromberg-Martin et al., 2010; Lammel et al., 2014; Roeper, 2013).”

“Although the specificity of the TH-GFP line has been questioned (Lammel et al., 2015), we found that GFP expression is specific for TH+ neurons in the SNc (Figure S3).”

“(legend continued on next page) 636 Cell 162, 635–647, July 30, 2015 ª2015 Elsevier Inc. to identify such connections (Chuhma et al., 2011; Xia et al., 2011) or found them to be very weak (Bocklisch et al., 2013).”

“Further development of high-throughput processing of these wholebrain datasets will be needed (Chung and Deisseroth, 2013; Kim et al., 2013, 2015) to fully capitalize on this opportunity for the rapid advancement of understanding structure-function relationships in the brain.”

“We injected CAV-cre into either the DMS or DLS and then injected AAVs expressing cre-dependent rabies glycoprotein (G) and TC, a high efficiency version of the TVA receptor fused to mCherry (Miyamichi et al., 2013), into the SNc.”

“(legend continued on next page) 636 Cell 162, 635–647, July 30, 2015 ª2015 Elsevier Inc. to identify such connections (Chuhma et al., 2011; Xia et al., 2011) or found them to be very weak (Bocklisch et al., 2013).”

“In fact, SNc DA neurons could in principle even be similar to VTA DA neurons in terms of the valence of information represented because mice will self-stimulate for SNc DA neuron activity just as for VTA DA neuron activity (Ilango et al., 2014; Rossi et al., 2013).”

“In TRIO, cre is delivered from the retrograde CAV-cre virus (Hnasko et al., 2006; Soudais et al., 2001), which transduces axon terminals and thereby defines ‘‘starter cells’’ (fromwhich rabies tracingwill occur) by virtue of their projection target.”

“Indeed, combined with other recent lines of evidence (Lammel et al., 2008, 2011; Margolis et al., 2008), we have found that projection-defined subpopulations of DA neurons display different intrinsic properties.”

“These regions of the dorsal striatum have been functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas might also participate i…”

“Similarly, for mice in which DA neuron TH expression was restricted to neurons projecting to either the DLS or the ventromedial striatum (VMS), DLS-projecting neurons were located more laterally than VMS-projecting neurons (Darvas and Palmiter, 2009; 2010; see also Schiemann et al., 2012).”

“In TRIO, cre is delivered from the retrograde CAV-cre virus (Hnasko et al., 2006; Soudais et al., 2001), which transduces axon terminals and thereby defines ‘‘starter cells’’ (fromwhich rabies tracingwill occur) by virtue of their projection target.”

“In fact, SNc DA neurons could in principle even be similar to VTA DA neurons in terms of the valence of information represented because mice will self-stimulate for SNc DA neuron activity just as for VTA DA neuron activity (Ilango et al., 2014; Rossi et al., 2013).”

“These regions of the dorsal striatum have been functionally distinguished by previous studies (Castañé et al., 2010; Faure et al., 2005; Featherstone and McDonald, 2004; Yin and Knowlton, 2004; Yin et al., 2004, 2005a, 2009, 2005b), leading us to hypothesize that SNc projections to these areas…”

“…which increase their firing in response to these stimuli, may signal salience rather than value (Brischoux et al., 2009; Bromberg-Martin et al., 2010; Matsumoto and Hikosaka, 2009), though controversy on this point remains (Cohen et al., 2012; Fiorillo et al., 2013, 2013; Ungless et al., 2004).”

“Similarly, for mice in which DA neuron TH expression was restricted to neurons projecting to either the DLS or the ventromedial striatum (VMS), DLS-projecting neurons were located more laterally than VMS-projecting neurons (Darvas and Palmiter, 2009; 2010; see also Schiemann et al., 2012).”

“Further development of high-throughput processing of these wholebrain datasets will be needed (Chung and Deisseroth, 2013; Kim et al., 2013, 2015) to fully capitalize on this opportunity for the rapid advancement of understanding structure-function relationships in the brain.”

“Similarly, for mice in which DA neuron TH expression was restricted to neurons projecting to either the DLS or the ventromedial striatum (VMS), DLS-projecting neurons were located more laterally than VMS-projecting neurons (Darvas and Palmiter, 2009; 2010; see also Schiemann et al., 2012).”

“A similar input-output architecture also applies to SNc-DA circuitry (Lerner et al., 2015 [in this issue of Cell]).”

“In vivo imaging with fiber photometry Fiber photometry was performed as in (28, 63, 64).”

“Those techniques are now starting to enable the streamlined analysis of axonal projections or cell positions in intact mouse brains in a manual or semi-automated way (Lerner et al., 2015; Menegas et al., 2015; Schwarz et al., 2015).”