Circuits in the brain possess a remarkable ability to orchestrate activities on different timescales, but how distinct circuits interact to sculpt diverse rhythms remains unresolved. The olfactory bulb is a classic example where slow, theta, and fast, gamma, rhythms coexist. Furthermore inhibitory interneurons generally implicated in rhythm generation are segregated into distinct layers, neatly separating local from global motifs. Here, combining intracellular recordings in vivo with circuit-specific optogenetic interference we dissect the contribution of inhibition to rhythmic activity in the mouse olfactory bulb. We found that the two inhibitory circuits control rhythms on distinct timescales: local, glomerular networks coordinate theta activity, regulating baseline and odor-evoked inhibition; granule cells orchestrate gamma synchrony and spike timing. Surprisingly, they did not contribute to baseline rhythms, or sniff-coupled odor-evoked inhibition despite their perceived dominance. Thus, activities on theta and gamma time scales are controlled by separate, dissociable inhibitory networks in the olfactory bulb.
Multiphoton imaging is widely used for recording activity simultaneously from many neurons in superficial cortical layers in vivo. Here we combine regenerative amplification multiphoton microscopy (RAMM) with genetically encoded calcium indicators to extend multiphoton imaging of neuronal population activity into layer 5 of adult mouse somatosensory cortex. We show that this approach can be used to record and quantify spontaneous and sensory-evoked activity in populations of layer 5 neuronal somata located as much as 800µm below the pia. In addition, we show that RAMM can be used to simultaneously image activity from large (~80) populations of apical dendrites and follow these dendrites down to their somata of origin.3
To identify the underlying reason for the controversial performance of tetracycline (Tet)-controlled regulated gene expression in mammalian neurons, we investigated each of the three components that comprise the Tet inducible systems, namely tetracyclines as inducers, tetracycline-transactivator (tTA) and reverse tTA (rtTA), and tTA-responsive promoters (Ptets). We have discovered that stably integrated Ptet becomes functionally silenced in the majority of neurons when it is inactive during development. Ptet silencing can be avoided when it is either not integrated in the genome or stably-integrated with basal activity. Moreover, long-term, high transactivator levels in neurons can often overcome integration-induced Ptet gene silencing, possibly by inducing promoter accessibility.
The raphe nuclei provide serotonergic innervation widely in the brain, thought to mediate a variety of neuromodulatory effects. The mammalian olfactory bulb (OB) is a prominent recipient of serotonergic fibers, particularly in the glomerular layer (GL), where they are thought to gate incoming signals from the olfactory nerve. The dorsal raphe nucleus (DRN) and the median raphe nucleus (MRN) are known to densely innervate the OB. The majority of such projections are thought to terminate in the GL, but this has not been explicitly tested. We sought to investigate this using recombinant adeno-associated viruses (rAAV)-mediated expression of green fluorescent protein (GFP)-synaptophysin targeted specifically to neurons of the DRN or the MRN. With DRN injections, labeled fibers were found mostly in the granule cell layer (GCL), not the GL. Conversely, dense labeling in the GL was observed with MRN injections, suggesting that the source of GL innervation is the MRN, not the DRN, as previously thought. The two raphe nuclei thus give dual innervation within the OB, with distinct innervation patterns. J. Comp. Neurol. 523:805–813, 2015. © 2015 Wiley Periodicals, Inc.
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