Teaching Neuroscience: Reviving Neuroanatomy, Notes on the 2022 Society for Neuroscience Professional Development Workshop on Teaching.

Students often find neuroanatomy a daunting exercise of rote memorization in a dead language. This workshop was designed to enliven the teaching of neuroanatomy. We recast the topic by extending it to the cellular and sub-cellular levels, animating it by learning to build a brain, and infusing the topic with the lively arts. Due to COVID's interference with the usual schedule of Society for Neuroscience (SfN) events, the 2021 Professional Development Workshop on Teaching was held as a webinar on April 12, 2022 with a follow-up question and answer session on June 7. In this workshop, not only were innovative teaching methods presented, but also the very definition of neuroanatomy was pushed to the limits-even reaching into the molecular and subcellular level. The presenters provided means of engaging students that were no cost, low cost, or well within the reach of most academic institutions. Judging by the attendance, this webinar was quite successful in its goals. Our speakers presented exciting and varied approaches to teaching neuroanatomy. Kaitlyn Casimo presented how the vast resources of the Allen Institute could be employed. Marc Nahmani described how open data resources could be utilized in creating a Course-Based Undergraduate Research Experience (CURE) on neural microanatomy. Erika Fanselow presented novel ways to overcome one of students' big hurdles in grasping neuroanatomy: understanding 3-D relationships. Len White described a creative approach in teaching neuroanatomy by incorporating the humanities, particularly art and literature. This article presents synopses of the presentations, which are written by the four presenters. Additionally, prompted by questions from the viewers, we have constructed a table of our favorite resources. A video of the original presentations as well as links to the subsequent Q & A sessions is available at https://neuronline.sfn.org/training/teaching-neuroscience-reviving-neuroanatomy/.

Target-specific M1 inputs to infragranular S1 pyramidal neurons.

The functional role of input from the primary motor cortex (M1) to primary somatosensory cortex (S1) is unclear; one key to understanding this pathway may lie in elucidating the cell-type specific microcircuits that connect S1 and M1. Recently, we discovered that a subset of pyramidal neurons in the infragranular layers of S1 receive especially strong input from M1 (Kinnischtzke AK, Simons DJ, Fanselow EE. Cereb Cortex 24: 2237-2248, 2014), suggesting that M1 may affect specific classes of pyramidal neurons differently. Here, using combined optogenetic and retrograde labeling approaches in the mouse, we examined the strengths of M1 inputs to five classes of infragranular S1 neurons categorized by their projections to particular cortical and subcortical targets. We found that the magnitude of M1 synaptic input to S1 pyramidal neurons varies greatly depending on the projection target of the postsynaptic neuron. Of the populations examined, M1-projecting corticocortical neurons in L6 received the strongest M1 inputs, whereas ventral posterior medial nucleus-projecting corticothalamic neurons, also located in L6, received the weakest. Each population also possessed distinct intrinsic properties. The results suggest that M1 differentially engages specific classes of S1 projection neurons, thereby regulating the motor-related influence S1 exerts over subcortical structures.

Neocortical somatostatin neurons reversibly silence excitatory transmission via GABAb receptors.

BACKGROUND: Understanding the dynamic range for excitatory transmission is a critical component of building a functional circuit diagram for the mammalian brain. Excitatory synaptic transmission is typically studied under optimized conditions, when background activity in the network is low. The range of synaptic function in the presence of inhibitory and excitatory activity within the neocortical circuit is unknown. RESULTS: Paired-cell recordings from pyramidal neurons in acute brain slices of mouse somatosensory cortex show that excitatory synaptic transmission is markedly suppressed during spontaneous network activity: EPSP amplitudes are 2-fold smaller and failure rates are greater than 50%. This suppression is mediated by tonic activation of presynaptic GABAb receptors gated by the spontaneous activity of somatostatin-expressing (Sst) interneurons. Optogenetic suppression of Sst neuron firing was sufficient to enhance EPSP amplitude and reduce failure rates, effects that were fully reversible and occluded by GABAb antagonists. CONCLUSIONS: These data indicate that Sst interneurons can rapidly and reversibly silence excitatory synaptic connections through the regulation of presynaptic release. This is an unanticipated role for Sst interneurons, which have been assigned a role only in fast GABAa-mediated inhibition. Because Sst interneuron activity has been shown to be regulated by sensory and motor input, these results suggest a mechanism by which functional connectivity and synaptic plasticity could be gated in a state-dependent manner.

Motor cortex broadly engages excitatory and inhibitory neurons in somatosensory barrel cortex.

Anatomical studies have shown that primary somatosensory (S1) and primary motor (M1) cortices are reciprocally connected. The M1 to S1 projection is thought to represent a modulatory signal that conveys motor-related information to S1. Here, we investigated M1 synaptic inputs to S1 by injecting an AAV virus containing channelrhodopsin-2 and a fluorescent tag into M1. Consistent with previous results, we found labeling of M1 axons within S1 that was most robust in the deep layers and in L1. Labeling was sparse in L4 and was concentrated in the interbarrel septa, largely avoiding barrel centers. In S1, we recorded in vitro from regular-spiking excitatory neurons and fast-spiking and somatostatin-expressing inhibitory interneurons. All 3 cell types had a high probability of receiving direct excitatory M1 input. Both excitatory and inhibitory cells within L4 were the least likely to receive such input from M1. Disynaptic inhibition was observed frequently, indicating that M1 recruits substantial inhibition within S1. Additionally, a subpopulation of L6 regular-spiking excitatory neurons received exceptionally strong M1 input. Overall, our results suggest that activation of M1 evokes within S1 a bombardment of excitatory and inhibitory synaptic activity that could contribute in a layer-specific manner to state-dependent changes in S1.

Central mechanisms of cranial nerve stimulation for epilepsy.

Stimulation of peripheral cranial nerves has been shown to exert anticonvulsant effects in animal models as well as in human patients. Specifically, stimulation of both the trigeminal and vagus nerves has been shown in multiple clinical trials to be anticonvulsant, and stimulation of these nerves at therapeutic levels does not cause pain or negatively affect brain function. However, the neuronal mechanisms by which such stimulation exerts therapeutic effects are not well understood. In this review, the possible locations of action for trigeminal nerve stimulation (TNS) and vagus nerve stimulation (VNS) are explored. Additionally, the multiple time scales on which TNS and VNS function are discussed.

Postnatal maturation of somatostatin-expressing inhibitory cells in the somatosensory cortex of GIN mice.

Postnatal inhibitory neuron development affects mammalian brain function, and failure of this maturation process may underlie pathological conditions such as epilepsy, schizophrenia, and depression. Furthermore, understanding how physiological properties of inhibitory neurons change throughout development is critical to understanding the role(s) these cells play in cortical processing. One subset of inhibitory neurons that may be affected during postnatal development is somatostatin-expressing (SOM) cells. A subset of these cells is labeled with green-fluorescent protein (GFP) in a line of mice known as the GFP-positive inhibitory neurons (GIN) line. Here, we studied how intrinsic electrophysiological properties of these cells changed in the somatosensory cortex of GIN mice between postnatal ages P11 and P32+. GIN cells were targeted for whole-cell current-clamp recordings and ranges of positive and negative current steps were presented to each cell. The results showed that as the neocortical circuitry matured during this critical time period multiple intrinsic and firing properties of GIN inhibitory neurons, as well as those of excitatory (regular-spiking [RS]) cells, were altered. Furthermore, these changes were such that the output of GIN cells, but not RS cells, increased over this developmental period. We quantified changes in excitability by examining the input-output relationship of both GIN and RS cells. We found that the firing frequency of GIN cells increased with age, while the rheobase current remained constant across development. This created a multiplicative increase in the input-output relationship of the GIN cells, leading to increases in gain with age. The input-output relationship of the RS cells, on the other hand, showed primarily a subtractive shift with age, but no substantial change in gain. These results suggest that as the neocortex matures, inhibition coming from GIN cells may become more influential in the circuit and play a greater role in the modulation of neocortical activity.

LTS and FS inhibitory interneurons, short-term synaptic plasticity, and cortical circuit dynamics.

Somatostatin-expressing, low threshold-spiking (LTS) cells and fast-spiking (FS) cells are two common subtypes of inhibitory neocortical interneuron. Excitatory synapses from regular-spiking (RS) pyramidal neurons to LTS cells strongly facilitate when activated repetitively, whereas RS-to-FS synapses depress. This suggests that LTS neurons may be especially relevant at high rate regimes and protect cortical circuits against over-excitation and seizures. However, the inhibitory synapses from LTS cells usually depress, which may reduce their effectiveness at high rates. We ask: by which mechanisms and at what firing rates do LTS neurons control the activity of cortical circuits responding to thalamic input, and how is control by LTS neurons different from that of FS neurons? We study rate models of circuits that include RS cells and LTS and FS inhibitory cells with short-term synaptic plasticity. LTS neurons shift the RS firing-rate vs. current curve to the right at high rates and reduce its slope at low rates; the LTS effect is delayed and prolonged. FS neurons always shift the curve to the right and affect RS firing transiently. In an RS-LTS-FS network, FS neurons reach a quiescent state if they receive weak input, LTS neurons are quiescent if RS neurons receive weak input, and both FS and RS populations are active if they both receive large inputs. In general, FS neurons tend to follow the spiking of RS neurons much more closely than LTS neurons. A novel type of facilitation-induced slow oscillations is observed above the LTS firing threshold with a frequency determined by the time scale of recovery from facilitation. To conclude, contrary to earlier proposals, LTS neurons affect the transient and steady state responses of cortical circuits over a range of firing rates, not only during the high rate regime; LTS neurons protect against over-activation about as well as FS neurons.

Trigeminal nerve stimulation: seminal animal and human studies for epilepsy and depression.

The unique ability to stimulate bilaterally, extracranially, and non-invasively may represent a significant advantage to invasive neuromodulation therapies. In humans thus far the technique has been applied noninvasively, and is termed external trigeminal nerve stimulation (eTNSTM).

The roles of somatostatin-expressing (GIN) and fast-spiking inhibitory interneurons in UP-DOWN states of mouse neocortex.

The neocortex contains multiple types of inhibitory neurons whose properties suggest they may play different roles within the cortical circuit. By recording from three cell types during two distinct network states (UP and DOWN states) in vitro, we were able to quantify differences in firing characteristics between these cells during different network regimes. We recorded from regular-spiking (RS) excitatory cells and two types of inhibitory neurons, the fast-spiking (FS) neurons and GFP- (and somatostatin-) expressing inhibitory neurons (GIN), in layer 2/3 of slices from mouse somatosensory neocortex. Comparisons of firing characteristics between these cells during UP- and DOWN-states showed several patterns. First, of these cell types, only GIN cells fired persistently during DOWN-states, whereas all three cell types fired readily during UP-states. Second, the onset of firing and distribution of action potentials throughout UP-states differed by cell type, showing that FS cell UP-state firing occurred preferentially near the beginning of the UP-state, whereas the firing of RS cells was slower to develop at the start of the UP-state, and GIN cell firing was sustained throughout the duration of the UP-state. Finally, membrane potential and spike correlations between heterogeneous cell types were more pronounced during UP-states and, in the case of RS synapses onto GIN cells, varied throughout the UP-state. These results suggest that there is a division of labor between FS and GIN cells as the UP-state progresses and suggest that GIN cells could be important in the termination of UP-states.

Selective, state-dependent activation of somatostatin-expressing inhibitory interneurons in mouse neocortex.

The specific functions of subtypes of cortical inhibitory neurons are not well understood. This is due in part to a dearth of information about the behaviors of interneurons under conditions when the surrounding circuit is in an active state. We investigated the firing behavior of a subset of inhibitory interneurons, identified using mice that express green fluorescent protein (GFP) in a subset of somatostatin-expressing inhibitory cells ("GFP-expressing inhibitory neuron" [GIN] cells). The somata of the GIN cells were in layer 2/3 of somatosensory cortex and had dense, layer 1-projecting axons that are characteristic of Martinotti neurons. Interestingly, GIN cells fired similarly during a variety of diverse activating conditions: when bathed in fluids with low-divalent cation concentrations, when stimulated with brief trains of local synaptic inputs, when exposed to group I metabotropic glutamate receptor agonists, or when exposed to muscarinic cholinergic receptor agonists. During these manipulations, GIN cells fired rhythmically and persistently in the theta-frequency range (3-10 Hz). Synchronous firing was often observed and its strength was directly proportional to the magnitude of electrical coupling between GIN cells. These effects were cell type specific: the four manipulations that persistently activated GIN cells rarely caused spiking of regular-spiking (RS) pyramidal cells or fast-spiking (FS) inhibitory interneurons. Our results suggest that supragranular GIN interneurons form an electrically coupled network that exerts a coherent 3- to 10-Hz inhibitory influence on its targets. Because GIN cells are more readily activated than RS and FS cells, it is possible that they act as "first responders" when cortical excitatory activity increases.

Navigating a sensorimotor loop.

Touch is an active process, but how do the body's somatic sensors influence its movement? In this issue of Neuron, Nguyen and Kleinfeld show that afferent activity from the whiskers on a rat's face trigger rapid and prolonged excitation of the motor neurons that drive movements of the same whiskers. Positive feedback through this sensorimotor loop may serve to optimize the interaction between sensors and stimuli.

Thalamocortical [correction of Thalamcortical] optimization of tactile processing according to behavioral state.

We propose a conceptual model that describes the operation of the main thalamocortical loop of the rat somatosensory system. According to this model, the asynchronous convergence of ascending and descending projections dynamically alters the physiological properties of thalamic neurons in the ventral posterior medial (VPM) nucleus as rats shift between three behavioral states. Two of these states are characterized by distinct modes of rhythmic whisker movements. We posit that these simultaneous shifts in exploratory behavioral strategy and in the physiological properties of VPM neurons allow rats to either (i) optimize the detection of stimuli that are novel or difficult to sense or (ii) process complex patterns of multi-whisker stimulation.

Dynamic shifting in thalamocortical processing during different behavioural states.

Recent experiments in our laboratory have indicated that as rats shift the behavioural strategy employed to explore their surrounding environment, there is a parallel change in the physiological properties of the neuronal ensembles that define the main thalamocortical loop of the trigeminal somatosensory system. Based on experimental evidence from several laboratories, we propose that this concurrent shift in behavioural strategy and thalamocortical physiological properties provides rats with an efficient way to optimize either the detection or analysis of complex tactile stimuli.