Single-unit Recordings Research Articles

A Modular Implementation to Handle and Benchmark Drift Correction for High-Density Extracellular Recordings.

High-density neural devices are now offering the possibility to record from neuronal populations in vivo at unprecedented scale. However, the mechanical drifts often observed in these recordings are currently a major issue for "spike sorting," an essential analysis step to identify the activity of single neurons from extracellular signals. Although several strategies have been proposed to compensate for such drifts, the lack of proper benchmarks makes it hard to assess the quality and effectiveness of motion correction. In this paper, we present a benchmark study to precisely and quantitatively evaluate the performance of several state-of-the-art motion correction algorithms introduced in the literature. Using simulated recordings with induced drifts, we dissect the origins of the errors performed while applying a motion correction algorithm as a preprocessing step in the spike sorting pipeline. We show how important it is to properly estimate the positions of the neurons from extracellular traces in order to correctly estimate the probe motion, compare several interpolation procedures, and highlight what are the current limits for motion correction approaches.

Dataset of human-single neuron activity during a Sternberg working memory task

We present a dataset of 1809 single neurons recorded from the human medial temporal lobe (amygdala and hippocampus) and medial frontal lobe (anterior cingulate cortex, pre-supplementary motor area, ventral medial prefrontal cortex) across 41 sessions from 21 patients that underwent seizure monitoring with depth electrodes. Subjects performed a screening task (907 neurons) to identify images for which highly selective cells were present. Subjects then performed a working memory task (902 neurons), in which they were sequentially presented with 1–3 images for which highly selective cells were present and, following a maintenance period, were asked if the probe was identical to one of the maintained images. This Neurodata Without Borders formatted dataset includes spike times, extracellular spike waveforms, stimuli presented, behavior, electrode locations, and subject demographics. As validation, we replicate previous findings on the selectivity of concept cells and their persistent activity during working memory maintenance. This large dataset of rare human single-neuron recordings and behavior enables the investigation of the neural mechanisms of working memory in humans.

Flexible, scalable, high channel count stereo-electrode for recording in the human brain

Over the past decade, stereotactically placed electrodes have become the gold standard for deep brain recording and stimulation for a wide variety of neurological and psychiatric diseases. Current electrodes, however, are limited in their spatial resolution and ability to record from small populations of neurons, let alone individual neurons. Here, we report on an innovative, customizable, monolithically integrated human-grade flexible depth electrode capable of recording from up to 128 channels and able to record at a depth of 10 cm in brain tissue. This thin, stylet-guided depth electrode is capable of recording local field potentials and single unit neuronal activity (action potentials), validated across species. This device represents an advance in manufacturing and design approaches which extends the capabilities of a mainstay technology in clinical neurology.

N-acetylcysteine attenuates accumbal core neuronal activity in response to morphine in the reinstatement of morphine CPP in morphine extinguished rats

Numerous studies have suggested that N-acetylcysteine (NAC), has the potential to suppress drug craving in people with substance use disorder and reduce drug-seeking behaviors in animals. The nucleus accumbens (NAc) plays a crucial role in the brain's reward system, with the nucleus accumbens core (NAcore) specifically implicated in compulsive drug seeking and relapse. In this study, we aimed to explore the impact of subchronic NAC administration during the extinction period and acute NAC administration on the electrical activity of NAcore neurons in response to a priming dose of morphine in rats subjected to extinction from morphine-induced place preference (CPP).We conducted single-unit recordings in anesthetized rats on the reinstatement day, following the establishment of morphine-induced conditioned place preference (7 mg/kg, s.c., 3 days), and subsequent drug-free extinction. In the subchronically NAC-treated groups, rats received daily injections of either NAC (50 mg/kg; i.p.) or saline during the extinction period. On the reinstatement day, we recorded the spontaneous activity of NAcore neurons for 15 min, administered a priming dose of morphine, and continued recording for an additional 45 min. While morphine excited most recorded neurons in saline-treated rats, it failed to alter firing rates in NAC-treated rats that had received NAC during the extinction period. For acutely NAC-treated animals, we recorded the baseline activity of NAcore neurons for 10 min before administering a single injection of either NAC (50 mg/kg; i.p.) or saline in rats with no treatment during the extinction. Following 30 min of recording and a priming dose of morphine (1 mg/kg, s.c.), the recording continued for an additional 30 min. The firing activity of NAcore neurons did not show significant changes after morphine or NAC injection. In conclusion, our findings emphasize that daily NAC administration during the extinction period significantly attenuates the morphine-induced increase in firing rates of NAcore neurons during the reinstatement of morphine CPP. However, acute NAC injection does not produce the same effect. These results suggest that modulating glutamate transmission through daily NAC during extinction may effectively inhibit the morphine place preference following the excitatory effects of morphine on NAcore neurons.

Human organotypic brain slice cultures: a detailed and improved protocol for preparation and long-term maintenance

The investigation of the human brain at cellular and microcircuit level remains challenging due to the fragile viability of neuronal tissue, inter- and intra-variability of the samples and limited availability of human brain material. Especially brain slices have proven to be an excellent source to investigate brain physiology and disease at cellular and small network level, overcoming the temporal limits of acute slices. Here we provide a revised, detailed protocol of the production and in-depth knowledge on long-term culturing of such human organotypic brain slice cultures for research purposes. We highlight the critical pitfalls of the culturing process of the human brain tissue and present exemplary results on viral expression, single-cell Patch-Clamp recordings, as well as multi-electrode array recordings as readouts for culture viability, enabling the use of organotypic brain slice cultures of these valuable tissue samples for basic neuroscience and disease modeling (Fig. 1).

Dopamine-integrated all-hydrogel multi-electrode arrays for neural activity recording.

Investigation of brain neural circuits is essential for deciphering the diagnostics and therapeutics of neurodegenerative diseases. The main concerns with traditional rigid metal electrodes include intrinsic mechanical mismatch between sensing electrodes and tissues, unavoidable foreign body responses, and inadequate spatiotemporal resolution, resulting in a deficiency of sensing performance. All-hydrogel neural electrodes with multi-electrode arrays (MEAs) suggest a viable way to modulate the trade-off between tissue-mechanical compliance and excellent spatiotemporal recording capacity, but still face the issues of insufficient conductivity and unstable interlayer bonding. Herein, we constructed a four-layer all-hydrogel neural electrode, by sandwiching a conductive hydrogel layer within two encapsulation hydrogel layers, with a shielding hydrogel layer located on top. We introduce a dual-strategy treatment to induce controllable phase separation in poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) hydrogel, which achieved ultra-high conductivity (up to 4176 S cm-1) comparable to that of metals and precise spatial resolution (∼15 μm) suitable for single neuron recording. In addition, the utilization of polyphenol chemistry mediated adaptive adhesion endowed this neural electrode with flexible and stable interlayer bonding among conductive-encapsulation-shielding layers and the tissue-electrode interface. Consequently, the all-hydrogel neural electrode exhibited a tenfold higher signal-to-noise ratio than a commercial silver electrode, realized the recording of weak neural activity signals within single and multiple neurons in epileptic rats, and applied man-made stimulation to the cerebral cortex of rats during seizures. This work provides a useful tool to understand the development, function and treatment of neurodegenerative diseases.

An Adaptive Leaky-Integrate and Firing Probability Model of an Electrically Stimulated Auditory Nerve Fiber.

Most neural models produce a spiking output and often represent the stochastic nature of the spike generation process via a stochastic output. Nonspiking neural models, on the other hand, predict the probability of a spike occurring in response to a stimulus. We propose a nonspiking model for an electrically stimulated auditory nerve fiber, which not only predicts the total probability of a spike occurring in response to a biphasic pulse but also the distribution of the spike time. Our adaptive leaky-integrate and firing probability (aLIFP) model can account for refractoriness, facilitation, accommodation, and long-term adaptation. All model parameters have been fitted to single cell recordings from electrically stimulated cat auditory nerve fibers. Afterward, the model was validated on recordings from auditory nerve fibers from cats and guinea pigs. The nonspiking nature of the model makes it fast and deterministic while still accounting for the stochastic nature of the spike generation process. Therefore, the relationship between the input to the model or model parameters and the model's output can be observed more directly than with stochastically spiking models.

Morphological disruption and visual tuning alterations in the primary visual cortex in glaucoma (DBA/2J) mice.

Glaucoma is a leading cause of irreversible blindness worldwide, and previous studies have shown that, in addition to affecting the eyes, it also causes abnormalities in the brain. However, it is not yet clear how the primary visual cortex (V1) is altered in glaucoma. This study used DBA/2J mice as a model for spontaneous secondary glaucoma. The aim of the study was to compare the electrophysiological and histomorphological characteristics of neurons in the V1 between 9-month-old DBA/2J mice and age-matched C57BL/6J mice. We conducted single-unit recordings in the V1 of light-anesthetized mice to measure the visually induced responses, including single-unit spiking and gamma band oscillations. The morphology of layer II/III neurons was determined by neuronal nuclear antigen staining and Nissl staining of brain tissue sections. Eighty-seven neurons from eight DBA/2J mice and eighty-one neurons from eight C57BL/6J mice were examined. Compared with the C57BL/6J group, V1 neurons in the DBA/2J group exhibited weaker visual tuning and impaired spatial summation. Moreover, fewer neurons were observed in the V1 of DBA/2J mice compared with C57BL/6J mice. These findings suggest that DBA/2J mice have fewer neurons in the V1 compared with C57BL/6J mice, and that these neurons have impaired visual tuning. Our findings provide a better understanding of the pathological changes that occur in V1 neuron function and morphology in the DBA/2J mouse model. This study might offer some innovative perspectives regarding the treatment of glaucoma.

Left Barrel Cortical Neurons Activity following Transplantation of Stem Cells into Right Lesioned-Barrel Cortex in Rats.

Stem cells (SCs) can improve the functional defects of brain injury. Rodents use their whiskers to get tactile information from their surroundings. The aim of this study was to investigate whether the transplantation of SCs into the lesioned barrel cortex can help neuronal function in the contralateral cortex. Sixteen male Wistar rats (200-230 g) were used in this experimental study. We induced a mechanical lesion in the right barrel cortex area of rats by removing this area by a 3 mm skin punch. Four groups containing one intact group of rats: group 1: control, and three lesion groups, group 2: lesion+un-differentiated dental pulp SCs (U-DPSCs), group 3: lesion+differentiated dental pulp SCs (D-DPSCs), and group 4: cell medium (vehicle) that were injected in the lesion area. Three weeks after transplantation of SCs or cell medium, the rats' responses of left barrel cortical neurons to controlled deflections of right whiskers were recorded by using the extracellular single-unit recordings technique. The results showed that the neural spontaneous activity and response magnitude of intact barrel cortex neurons in the lesion group decreased significantly (P<0.05) compared to the control group while ON and OFF responses were improved in the D-DPSCs (P<0.001) group compared to the vehicle group three weeks after transplantation. Transplantation of dental pulp mesenchymal SCs significantly improved the neural responses of the left barrel cortex that was depressed in the vehicle group.

Pulse-width modulated temporal interference (PWM-TI) brain stimulation

BackgroundElectrical stimulation involving temporal interference of two different kHz frequency sinusoidal electric fields (temporal interference (TI)) enables non-invasive deep brain stimulation, by creating an electric field that is amplitude modulated at the slow difference frequency (within the neural range), at the target brain region. ObjectiveHere, we investigate temporal interference neural stimulation using square, rather than sinusoidal, electric fields that create an electric field that is pulse-width, but not amplitude, modulated at the difference frequency (pulse-width modulated temporal interference, (PWM-TI)). Methods/ResultsWe show, using ex-vivo single-cell recordings and in-vivo calcium imaging, that PWM-TI effectively stimulates neural activity at the difference frequency at a similar efficiency to traditional TI. We then demonstrate, using computational modelling, that the PWM stimulation waveform induces amplitude-modulated membrane potential depolarization due to the membrane's intrinsic low-pass filtering property. ConclusionsPWM-TI can effectively drive neural activity at the difference frequency. The PWM-TI mechanism involves converting an envelope amplitude-fixed PWM field to an amplitude-modulated membrane potential via the low-pass filtering of the passive neural membrane. Unveiling the biophysics underpinning the neural response to complex electric fields may facilitate the development of new brain stimulation strategies with improved precision and efficiency.

Soft high-density neural probes enable stable single-neuron recordings.

Soft high-density neural probes enable stable single-neuron recordings.

Time-resolved single-cell transcriptomics defines immune trajectories in glioblastoma

Deciphering the cell-state transitions underlying immune adaptation across time is fundamental for advancing biology. Empirical invivo genomic technologies that capture cellular dynamics are currently lacking. We present Zman-seq, a single-cell technology recording transcriptomic dynamics across time by introducing time stamps into circulating immune cells, tracking them in tissues for days. Applying Zman-seq resolved cell-state and molecular trajectories of the dysfunctional immune microenvironment in glioblastoma. Within 24hours of tumor infiltration, cytotoxic natural killer cells transitioned to a dysfunctional program regulated by TGFB1 signaling. Infiltrating monocytes differentiated into immunosuppressive macrophages, characterized by the upregulation of suppressive myeloid checkpoints Trem2, Il18bp, and Arg1, over 36 to 48hours. Treatment with an antagonistic anti-TREM2 antibody reshaped the tumor microenvironment by redirecting the monocyte trajectory toward pro-inflammatory macrophages. Zman-seq is a broadly applicable technology, enabling empirical measurements of differentiation trajectories, which can enhance the development of more efficacious immunotherapies.

Activity-dependent recruitment of inhibition and excitation in the awake mammalian cortex during electrical stimulation

Electrical stimulation is an effective tool for mapping and altering brain connectivity, with applications ranging from treating pharmacology-resistant neurological disorders to providing sensory feedback for neural prostheses. Paramount to the success of these applications is the ability to manipulate electrical currents to precisely control evoked neural activity patterns. However, little is known about stimulation-evoked responses in inhibitory neurons nor how stimulation-evoked activity patterns depend on ongoing neural activity. In this study, we used 2-photon imaging and cell-type specific labeling to measure single-cell responses of excitatory and inhibitory neurons to electrical stimuli in the visual cortex of awake mice. Our data revealed strong interactions between electrical stimulation and pre-stimulus activity of single neurons in awake animals and distinct recruitment and response patterns for excitatory and inhibitory neurons. This work demonstrates the importance of cell-type-specific labeling of neurons in future studies.

Stable coding of aversive associations in medial prefrontal populations.

The medial prefrontal cortex (mPFC) is at the core of numerous psychiatric conditions, including fear and anxiety-related disorders. Whereas an abundance of evidence suggests a crucial role of the mPFC in regulating fear behaviour, the precise role of the mPFC in this process is not yet entirely clear. While studies at the single-cell level have demonstrated the involvement of this area in various aspects of fear processing, such as the encoding of threat-related cues and fear expression, an increasingly prevalent idea in the systems neuroscience field is that populations of neurons are, in fact, the essential unit of computation in many integrative brain regions such as prefrontal areas. What mPFC neuronal populations represent when we face threats? To address this question, we performed electrophysiological single-unit population recordings in the dorsal mPFC while mice faced threat-predicting cues eliciting defensive behaviours, and performed pharmacological and optogenetic inactivations of this area and the amygdala. Our data indicated that the presence of threat-predicting cues induces a stable coding dynamics of internally driven representations in the dorsal mPFC, necessary to drive learned defensive behaviours. Moreover, these neural population representations primary reflect learned associations rather than specific defensive behaviours, and the construct of such representations relies on the functional integrity of the amygdala.

Analysis of cellular response to drugs with a microfluidic single-cell platform based on hyperspectral imaging

BackgroundCellular response to pharmacological action of drugs is significant for drug development. Traditional detection method for cellular response to drugs normally rely on cell proliferation assay and metabolomics examination. In principle, these analytical methods often required cell labeling, invasion analysis, and hours of co-culture with drugs, which are relatively complex and time-consuming. Moreover, these methods can only indicate the drug effectiveness on cell colony rather than single cells. Thus, to meet the requirements of personal precision medicine, the development of drug response analysis on the high resolution of single cell is demanded. ResultsTo provide precise result for drug response on single-cell level, a microfluidic platform coupled with the label-free hyperspectral imaging was developed. With the help of horizontal single-cell trapping sieves, hundreds of single cells were trapped independently in microfluidic channels for the purposes of real-time drug delivery and single-cell hyperspectral image recording. To significantly identify the cellular hyperspectral change after drug stimulation, the differenced single-cell spectrum was proposed. Compared with the deep learning classification method based on hyperspectral images, an optimal performance can be achieved by the classification strategy based on differenced spectra. And the cellular response to different reagents, for example, K+, Epidermal Growth Factor (EGF), and Gefitinib at different concentrations can be accurately characterized by the differenced single-cell spectra analysis. Significance and noveltyThe high-throughput, rapid analysis of cellular response to drugs at the single-cell level can be accurately performed by our platform. After systematically analyzing the materials and the structures of the single-cell microfluidic chip, the optimal single-cell trapping method was proposed to contribute to the further application of hyperspectral imaging on microfluidic single-cell analysis. And the hyperspectral characterization of single-cell with cancer drug stimulation proved the application potential of our method in personal cancer medication.

Subthalamic nucleus neuronal activity entropy in the effective stimulation area in patients with Parkinson’s disease

One of the most successful and promising treatments for Parkinson’s disease today is deep brain stimulation (DBS). Accurate localization of the stimulation area is critical for the outcome of surgical DBS electrode implantation in the subthalamic nucleus (STN). To achieve this, microelectrode recording is used during surgery to precisely locate the STN borders [1], while test stimulations with a macroelectrode are widely employed to enhance DBS electrode placement accuracy. Thus, through testing multiple trajectories, the most clinically effective one can be selected. However, a comprehensive and reliable description of the specific single neuron activity associated with successful DBS electrode implantation is currently lacking. A study was conducted using microelectrode recordings (MER) of single neuron activity in the STN of 21 Parkinson’s disease patients (UPDRS III off/on=46.9/12.8) to identify the neuronal activity features associated with the most favorable clinical outcome of test stimulation. A comparative analysis of 29 activity parameters, such as firing rate, oscillatory activity intensity in different frequency ranges (Oscores), coefficient of variation, burst, pause index, among others [2], was conducted for 618 identified neurons. In addition, the approach of computing the entropy of the change in interspike intervals (ISI) of the pulse sequence [3] and the process of identifying patterns of neural activity using hierarchical clustering [4] were implemented. A comparative analysis of single neuron activity indicated notable (p 0.05) variations between disregarded and selected paths for the ultimate implantation of DBS electrode, solely relying on entropy parameters. Trajectories that delivered optimal test stimulation outcomes demonstrated reduced entropy of interspike intervals. Only the patterns of neuronal activity characterized by extended periods of steady firing and brief pauses, referred to as tonic and pause activity, respectively, were found to contribute to the difference in entropy between trajectories. In contrast, there were no statistically significant differences detected in entropy values associated with neurons demonstrating burst activity. Furthermore, we observed a positive and significant correlation between entropy values and the degree of improvement in disease presentation before and after administration of medication. A comparative analysis was conducted on the activity of single neurons along different trajectories with varied test stimulation responses. The results showed a weak effectiveness of linear parameters in determining the optimal electrode insertion path for improved clinical outcomes. However, the use of non-linear activity parameters, specifically entropy, in single neurons effectively differentiated trajectories with significant versus absent/insignificant test stimulation results. Furthermore, the significance of entropy in establishing the basal ganglia functions and describing information transfer processes in movement control is emphasized by interpreting this parameter as a measure of uncertainty or unpredictability of the information system in relation to the findings of other studies [5].

Соотношение возбудительных и тормозных синаптических процессов в антиноцицептивных ядрах PAG и RMG в их реципрокных взаимоотношениях в интактном препарате с воздействием гидрокортизона

Microelectrophysiological studies were carried out on 12 intact Albino rats (250±30 g) with hydrocortisone injection, compared with the norm. A software mathematical analysis of a single spike activity of 282 neurons was carried out. The spike activity of single PAG and RMG neurons was recorded extracellularly in separate series of experiments, with successive involvement of each of them as stimulated and recorded. When assessing the relative degree severity of depressor and excitatory post-stimulus effects, using the example of diagrams of the average spike frequency in PAG at HFS RMG under hydrocortisone exposure in normal conditions, both in the depressor and excitatory sequences, in comparison with those without hydrocortisone, no significant changes were detected, but there was a greater severity of these values in the norm without hydrocortisone, in particular, preceding excitatory effects, which indicates not only the harmlessness of the indicated dose of hydrocortisone, but also a certain improvement in the normal status. In RMG neurons on HFS PAG, both in the norm without and under hydrocortisone exposure, a protective increase in tetanic depression was revealed in both sequences, without excess of poststimulus excitability. In general, for both PAG and RMG, a greater severity of pre- and post-stimulus frequency, preceding and accompanying excitatory effects in the norm without hydrocortisone, and a greater balance of depressor and excitatory post-stimulus manifestations of activity under hydrocortisone exposure than in the norm were revealed, which indicates the safety and even the benefit of that dose of hydrocortisone.

Holistic bursting cells store long-term memory in auditory cortex

The sensory neocortex has been suggested to be a substrate for long-term memory storage, yet which exact single cells could be specific candidates underlying such long-term memory storage remained neither known nor visible for over a century. Here, using a combination of day-by-day two-photon Ca2+ imaging and targeted single-cell loose-patch recording in an auditory associative learning paradigm with composite sounds in male mice, we reveal sparsely distributed neurons in layer 2/3 of auditory cortex emerged step-wise from quiescence into bursting mode, which then invariably expressed holistic information of the learned composite sounds, referred to as holistic bursting (HB) cells. Notably, it was not shuffled populations but the same sparse HB cells that embodied the behavioral relevance of the learned composite sounds, pinpointing HB cells as physiologically-defined single-cell candidates of an engram underlying long-term memory storage in auditory cortex.

A Nanozyme-Based Electrode for High-Performance Neural Recording.

Implanted neural electrodes have been widely used to treat brain diseases that require high sensitivity and biocompatibility at the tissue-electrode interface. However, currently used clinical electrodes cannot meet both these requirements simultaneously, which hinders the effective recording of electronic signals. Herein, nanozyme-based neural electrodes incorporating bioinspired atomically precise clusters are developed as a general strategy with a heterogeneous design for multiscale and ultrasensitive neural recording via quantum transport and biocatalytic processes. Owing to the dual high-speed electronic and ionic currents at the electrode-tissue interface, the impedance of nanozyme electrodes is 26 times lower than that of state-of-the-art metal electrodes, and the acquisition sensitivity for the local field potential is ≈10 times higher than that of clinical PtIr electrodes, enabling a signal-to-noise ratio (SNR) of up to 14.7dB for single-neuron recordings in rats. The electrodes provide more than 100-fold higher antioxidant and multi-enzyme-like activities, which effectively decrease 67% of the neuronal injury area by inhibiting glial proliferation and allowing sensitive and stable neural recording. Moreover, nanozyme electrodes can considerably improve the SNR of seizures in acute epileptic rats and are expected to achieve precise localization of seizure foci in clinical settings.

Prelimbic and infralimbic responsivity to amygdala input is modified by gonadal hormones in parallel to low anxiety-like behavior in ovariectomized rats

Gonadal hormones may influence sexual activity by reducing anxiety. The basolateral amygdala (BLA) and prelimbic (PL) and infralimbic (IL) cortical regions comprise a loop that is related to fear, anxiety, and social behavior. In female ovariectomized rats, actions of estradiol, progesterone, and sequential estradiol and progesterone administration were explored in the open field test (OFT) and plus maze test (PMT) to evaluate signs of anxiety-like behavior. The three hormonal treatments reduced indicators of anxiety in the PMT but did not influence behavior in the OFT. In the same behaviorally tested rats under urethane anesthesia, single-unit extracellular recordings were obtained from the PL and IL during electrical stimulation of the BLA. The analysis of 250 ms peristimulus histograms showed that BLA stimulation produced two kinds of response. A small group of neurons increased their firing rate after BLA stimulation. Most neurons exhibited a reduction of spiking. Neurons that increased their firing rate after BLA stimulation did not show any difference with the hormonal treatments. In neurons that were inhibited by BLA stimulation, estradiol reduced the neuronal firing rate in the PL and IL, and progesterone alone and the sequential administration of estradiol followed by progesterone administration 24 h later (priming) increased the firing rate during the 240 ms before BLA stimulation. Analyses of responsivity of the PL and IL during electrical stimulation of the BLA indicated that estradiol, progesterone, and estradiol followed by progesterone administration 24 h later (priming) reduced inhibitory actions of the BLA on the PL but not IL. In the BLA-IL connection, progesterone exacerbated the inhibitory response. These findings indicate that anxiolytic actions of estradiol, progesterone, and estradiol followed by progesterone administration 24 h later (priming) correspond to lower BLA-PL responsivity. Actions of progesterone on BLA-IL responsivity appear to contribute to sexual activity by interacting with other forebrain structures that are also related to sexual receptivity.