Simultaneous mesoscopic and two-photon imaging of neuronal activity in cortical circuits.

Spontaneous and sensory-evoked activity propagates across varying spatial scales in the mammalian cortex, but technical challenges have limited conceptual links between the function of local neuronal circuits and brain-wide network dynamics. We present a method for simultaneous cellular-resolution two-photon calcium imaging of a local microcircuit and mesoscopic widefield calcium imaging of the entire cortical mantle in awake mice. Our multi-scale approach involves a microscope with an orthogonal axis design where the mesoscopic objective is oriented above the brain and the two-photon objective is oriented horizontally, with imaging performed through a microprism. We also introduce a viral transduction method for robust and widespread gene delivery in the mouse brain. These approaches allow us to identify the behavioral state-dependent functional connectivity of pyramidal neurons and vasoactive intestinal peptide-expressing interneurons with long-range cortical networks. Our imaging system provides a powerful strategy for investigating cortical architecture across a wide range of spatial scales

Targeted optogenetic stimulation and recording of neurons in vivo using cell-type-specific expression of Channelrhodopsin-2

A major long-term goal of systems neuroscience is to identify the different roles of neural subtypes in brain circuit function. The ability to causally manipulate selective cell types is critical to meeting this goal. This protocol describes techniques for optically stimulating specific populations of excitatory neurons and inhibitory interneurons in vivo in combination with electrophysiology. Cell type selectivity is obtained using Cre-dependent expression of the light-activated channel Channelrhodopsin-2. We also describe approaches for minimizing optical interference with simultaneous extracellular and intracellular recording. These optogenetic techniques provide a spatially and temporally precise means of studying neural activity in the intact brain and allow a detailed examination of the effect of evoked activity on the surrounding local neural network. Injection of viral vectors requires 30–45 min, and in vivo electrophysiology with optogenetic stimulation requires 1–4 h.National Institutes of Health (U.S.)National Science Foundation (U.S.)Simons FoundationNational Institutes of Health (U.S.). Pioneer AwardNational Eye Institue (K99 Award)Knut and Alice Wallenberg Foundation (Postdoctoral Fellowship)Brain & Behavior Research Foundation. Young Investigator AwardThomas F. Peterse

Therapeutic Use of Self and Fieldwork Experience: An Exploration of the Art and Science of Occupational Therapy

The clinical practice of occupational therapy has been described as a blend of both art and science. For occupational therapy students, Level II fieldwork experiences offer early opportunities to refine both client-centered attitudes and scientific aptitude in relationship-based caregiving. In this retrospective study, researchers examined the ability to predict final Fieldwork Performance Evaluation scores from the following non-cognitive (i.e., art) and cognitive (i.e., science) variables: ranked student responses to the Self-Assessment of Modes Questionnaire (v.II); undergraduate grade point average (GPA; cumulative and science), and Graduate Record Examination (GRE) scores (quantitative, verbal, and analytic). Using a series of simple linear regressions, researchers analyzed data from sixty-nine master’s-level occupational therapy students. For the first Level II fieldwork experience, empathizing and empathizing-revised modes appeared to be a significant predictor with moderate, positive correlation coefficients (p=.008, r=.329; p=.01, r=.296, respectively). For the second Level II fieldwork experience, collaborating and instructing modes appeared to be significant predictors (p=.036, r= -.255; p=.037, r=.254 respectively). GPA and GRE scores were not predictive of fieldwork success. The degree to which art and science shape expectations for relationship-based client interactions during fieldwork experiences requires further investigation. However, calling attention to occupational therapy students’ preferred communication modes highlight how client interactions may be shaped to fit the students’ natural tendencies rather than the needs of the client

Value of Case Studies for Translating Evidence in Occupational Therapy: A New 3-Dimensional Model

Importance: Occupational therapy (OT) practitioners often address complex occupational performance needs of diverse consumers. Professional reasoning, theory, and trial and error interventions can be demonstrated in case studies. Case studies are usually relegated to the lowest level in evidence hierarchies. Yet they have a long tradition in health care literature, and may particularly document the holism, inclusivity, uniqueness, and complexity of OT practice. Objective: To determine how case studies make a distinct contribution to professional knowledge, using a new model for categorizing them. Evidence Review: 103 case studies published in AJOT from 1990-2023 were rated by the authors “1” or “2” on features derived from 3 axes: (1) frequency such a case would be encountered in current professional practice (rare = 2), (2) complexity of the causes of the client’s occupational challenges (complex = 2), and (3) complexity of the OT intervention (multidimensional = 2). Findings: 93.2% of case studies had some feature of novelty or complexity: 35.9% portrayed a rare/novel diagnosis, 63.1% displayed complex problem causes, and 83.5% involved multi-dimensional OT interventions. In 55.3% of the articles both the causes and the interventions were complex. Conclusions and Relevance: The predominance of novelty/complexity among the articles supports the claim that case studies mostly describe professional decisions and outcomes in those therapy situations that are less amenable to group experimental studies (infrequent/novel, complex causes, complex interventions). The value of the 3-axis model of case studies was supported. Case studies are a distinct source of evidence supporting inclusive, client-centered practice

Noninvasive optical inhibition with a red-shifted microbial rhodopsin

Optogenetic inhibition of the electrical activity of neurons enables the causal assessment of their contributions to brain functions. Red light penetrates deeper into tissue than other visible wavelengths. We present a red-shifted cruxhalorhodopsin, Jaws, derived from Haloarcula (Halobacterium) salinarum (strain Shark) and engineered to result in red light–induced photocurrents three times those of earlier silencers. Jaws exhibits robust inhibition of sensory-evoked neural activity in the cortex and results in strong light responses when used in retinas of retinitis pigmentosa model mice. We also demonstrate that Jaws can noninvasively mediate transcranial optical inhibition of neurons deep in the brains of awake mice. The noninvasive optogenetic inhibition opened up by Jaws enables a variety of important neuroscience experiments and offers a powerful general-use chloride pump for basic and applied neuroscience.McGovern Institute for Brain Research at MIT (Razin Fellowship)United States. Defense Advanced Research Projects Agency. Living Foundries Program (HR0011-12-C-0068)Harvard-MIT Joint Research Grants Program in Basic NeuroscienceHuman Frontier Science Program (Strasbourg, France)Institution of Engineering and Technology (A. F. Harvey Prize)McGovern Institute for Brain Research at MIT. Neurotechnology (MINT) ProgramNew York Stem Cell Foundation (Robertson Investigator Award)National Institutes of Health (U.S.) (New Innovator Award 1DP2OD002002)National Institute of General Medical Sciences (U.S.) (EUREKA Award 1R01NS075421)National Institutes of Health (U.S.) (Grant 1R01DA029639)National Institutes of Health (U.S.) (Grant 1RC1MH088182)National Institutes of Health (U.S.) (Grant 1R01NS067199)National Science Foundation (U.S.) (Career Award CBET 1053233)National Science Foundation (U.S.) (Grant EFRI0835878)National Science Foundation (U.S.) (Grant DMS0848804)Society for Neuroscience (Research Award for Innovation in Neuroscience)Wallace H. Coulter FoundationNational Institutes of Health (U.S.) (RO1 MH091220-01)Whitehall FoundationEsther A. & Joseph Klingenstein Fund, Inc.JPB FoundationPIIF FundingNational Institute of Mental Health (U.S.) (R01-MH102441-01)National Institutes of Health (U.S.) (DP2-OD-017366-01)Massachusetts Institute of Technology. Simons Center for the Social Brai

Rapid fluctuations in functional connectivity of cortical networks encode spontaneous behavior

Experimental work across species has demonstrated that spontaneously generated behaviors are robustly coupled to variations in neural activity within the cerebral cortex. Functional magnetic resonance imaging data suggest that temporal correlations in cortical networks vary across distinct behavioral states, providing for the dynamic reorganization of patterned activity. However, these data generally lack the temporal resolution to establish links between cortical signals and the continuously varying fluctuations in spontaneous behavior observed in awake animals. Here, we used wide-field mesoscopic calcium imaging to monitor cortical dynamics in awake mice and developed an approach to quantify rapidly time-varying functional connectivity. We show that spontaneous behaviors are represented by fast changes in both the magnitude and correlational structure of cortical network activity. Combining mesoscopic imaging with simultaneous cellular-resolution two-photon microscopy demonstrated that correlations among neighboring neurons and between local and large-scale networks also encode behavior. Finally, the dynamic functional connectivity of mesoscale signals revealed subnetworks not predicted by traditional anatomical atlas-based parcellation of the cortex. These results provide new insights into how behavioral information is represented across the neocortex and demonstrate an analytical framework for investigating time-varying functional connectivity in neural networks

Driving fast-spiking cells induces gamma rhythm and controls sensory responses,”

Cortical gamma oscillations (20280 Hz) predict increases in focused attention, and failure in gamma regulation is a hallmark of neurological and psychiatric disease. Current theory predicts that gamma oscillations are generated by synchronous activity of fast-spiking inhibitory interneurons, with the resulting rhythmic inhibition producing neural ensemble synchrony by generating a narrow window for effective excitation. We causally tested these hypotheses in barrel cortex in vivo by targeting optogenetic manipulation selectively to fast-spiking interneurons. Here we show that light-driven activation of fast-spiking interneurons at varied frequencies (82200 Hz) selectively amplifies gamma oscillations. In contrast, pyramidal neuron activation amplifies only lower frequency oscillations, a cell-type-specific double dissociation. We found that the timing of a sensory input relative to a gamma cycle determined the amplitude and precision of evoked responses. Our data directly support the fast-spiking-gamma hypothesis and provide the first causal evidence that distinct network activity states can be induced in vivo by cell-type-specific activation. Brain states characterized by rhythmic electrophysiological activity have been studied intensively for more than 80 years Cell-type-specific expression of channelrhodopsin-2 To test directly the hypothesis that FS interneuron activity in an in vivo cortical circuit is sufficient to induce gamma oscillations, we used the light-sensitive bacteriorhodopsin Chlamydomonas reinhardtii channelrhodopsin-2 (ChR2), a cation channel activated by ,470 nm blue ligh

Rapid Effects of Hearing Song on Catecholaminergic Activity in the Songbird Auditory Pathway

Catecholaminergic (CA) neurons innervate sensory areas and affect the processing of sensory signals. For example, in birds, CA fibers innervate the auditory pathway at each level, including the midbrain, thalamus, and forebrain. We have shown previously that in female European starlings, CA activity in the auditory forebrain can be enhanced by exposure to attractive male song for one week. It is not known, however, whether hearing song can initiate that activity more rapidly. Here, we exposed estrogen-primed, female white-throated sparrows to conspecific male song and looked for evidence of rapid synthesis of catecholamines in auditory areas. In one hemisphere of the brain, we used immunohistochemistry to detect the phosphorylation of tyrosine hydroxylase (TH), a rate-limiting enzyme in the CA synthetic pathway. We found that immunoreactivity for TH phosphorylated at serine 40 increased dramatically in the auditory forebrain, but not the auditory thalamus and midbrain, after 15 min of song exposure. In the other hemisphere, we used high pressure liquid chromatography to measure catecholamines and their metabolites. We found that two dopamine metabolites, dihydroxyphenylacetic acid and homovanillic acid, increased in the auditory forebrain but not the auditory midbrain after 30 min of exposure to conspecific song. Our results are consistent with the hypothesis that exposure to a behaviorally relevant auditory stimulus rapidly induces CA activity, which may play a role in auditory responses