Robert A. A. Campbell

Cellular-resolution population imaging reveals robust sparse coding in the Drosophila mushroom body.

Sensory stimuli are represented in the brain by the activity of populations of neurons. In most biological systems, studying population coding is challenging since only a tiny proportion of cells can be recorded simultaneously. Here we used two-photon imaging to record neural activity in the relatively simple Drosophila mushroom body (MB), an area involved in olfactory learning and memory. Using the highly sensitive calcium indicator GCaMP3, we simultaneously monitored the activity of >100 MB neurons in vivo (∼5% of the total population). The MB is thought to encode odors in sparse patterns of activity, but the code has yet to be explored either on a population level or with a wide variety of stimuli. We therefore imaged responses to odors chosen to evaluate the robustness of sparse representations. Different odors activated distinct patterns of MB neurons; however, we found no evidence for spatial organization of neurons by either response probability or odor tuning within the cell body layer. The degree of sparseness was consistent across a wide range of stimuli, from monomolecular odors to artificial blends and even complex natural smells. Sparseness was mainly invariant across concentrations, largely because of the influence of recent odor experience. Finally, in contrast to sensory processing in other systems, no response features distinguished natural stimuli from monomolecular odors. Our results indicate that the fundamental feature of odor processing in the MB is to create sparse stimulus representations in a format that facilitates arbitrary associations between odor and punishment or reward.

Physiological and behavioral studies of spatial coding in the auditory cortex.

Despite extensive subcortical processing, the auditory cortex is believed to be essential for normal sound localization. However, we still have a poor understanding of how auditory spatial information is encoded in the cortex and of the relative contribution of different cortical areas to spatial hearing. We investigated the behavioral consequences of inactivating ferret primary auditory cortex (A1) on auditory localization by implanting a sustained release polymer containing the GABA(A) agonist muscimol bilaterally over A1. Silencing A1 led to a reversible deficit in the localization of brief noise bursts in both the horizontal and vertical planes. In other ferrets, large bilateral lesions of the auditory cortex, which extended beyond A1, produced more severe and persistent localization deficits. To investigate the processing of spatial information by high-frequency A1 neurons, we measured their binaural-level functions and used individualized virtual acoustic space stimuli to record their spatial receptive fields (SRFs) in anesthetized ferrets. We observed the existence of a continuum of response properties, with most neurons preferring contralateral sound locations. In many cases, the SRFs could be explained by a simple linear interaction between the acoustical properties of the head and external ears and the binaural frequency tuning of the neurons. Azimuth response profiles recorded in awake ferrets were very similar and further analysis suggested that the slopes of these functions and location-dependent variations in spike timing are the main information-bearing parameters. Studies of sensory plasticity can also provide valuable insights into the role of different brain areas and the way in which information is represented within them. For example, stimulus-specific training allows accurate auditory localization by adult ferrets to be relearned after manipulating binaural cues by occluding one ear. Reversible inactivation of A1 resulted in slower and less complete adaptation than in normal controls, whereas selective lesions of the descending cortico collicular pathway prevented any improvement in performance. These results reveal a role for auditory cortex in training-induced plasticity of auditory localization, which could be mediated by descending cortical pathways.

Mice Lacking the ITIM-Containing Receptor G6b-B Exhibit Macrothrombocytopenia and Aberrant Platelet Function

An inhibitory receptor ensures that megakaryocytes produce proper numbers of functional platelets. Controlling Platelet Production Megakaryocytes reside in the bone marrow, where they produce platelets, cell fragments that form clots to prevent blood loss at sites of damage to the vasculature. Platelets and megakaryocytes share many activating receptors on their surface, but unlike platelets, megakaryocytes fail to become activated when exposed to components of the extracellular matrix. Mazharian et al. found that mice deficient in the immunoreceptor tyrosine–based inhibition motif–containing receptor G6b-B had fewer and larger platelets than did their wild-type counterparts. In addition, G6b-B–deficient mice exhibited increased bleeding in response to damage and had activated megakaryocytes, which resulted in the production of defective platelets. Together, these data suggest that G6b-B dampens activating signals in megakaryocytes to enable the generation of the appropriate number of functional platelets. Platelets are highly reactive cell fragments that adhere to exposed extracellular matrix (ECM) and prevent excessive blood loss by forming clots. Paradoxically, megakaryocytes, which produce platelets in the bone marrow, remain relatively refractory to the ECM-rich environment of the bone marrow despite having the same repertoire of receptors as platelets. These include the ITAM (immunoreceptor tyrosine–based activation motif)–containing collagen receptor complex, which consists of glycoprotein VI (GPVI) and the Fc receptor γ-chain, and the ITIM (immunoreceptor tyrosine–based inhibition motif)–containing receptor G6b-B. We showed that mice lacking G6b-B exhibited macrothrombocytopenia (reduced platelet numbers and the presence of enlarged platelets) and a susceptibility to bleeding as a result of aberrant platelet production and function. Platelet numbers were markedly reduced in G6b-B–deficient mice compared to those in wild-type mice because of increased platelet turnover. Furthermore, megakaryocytes in G6b-B–deficient mice showed enhanced metalloproteinase production, which led to increased shedding of cell-surface receptors, including GPVI and GPIbα. In addition, G6b-B–deficient megakaryocytes exhibited reduced integrin-mediated functions and defective formation of proplatelets, the long filamentous projections from which platelets bud off. Together, these findings establish G6b-B as a major inhibitory receptor regulating megakaryocyte activation, function, and platelet production.

Virtual Adult Ears Reveal the Roles of Acoustical Factors and Experience in Auditory Space Map Development

Auditory neurons in the superior colliculus (SC) respond preferentially to sounds from restricted directions to form a map of auditory space. The development of this representation is shaped by sensory experience, but little is known about the relative contribution of peripheral and central factors to the emergence of adult responses. By recording from the SC of anesthetized ferrets at different age points, we show that the map matures gradually after birth; the spatial receptive fields (SRFs) become more sharply tuned and topographic order emerges by the end of the second postnatal month. Principal components analysis of the head-related transfer function revealed that the time course of map development is mirrored by the maturation of the spatial cues generated by the growing head and external ears. However, using virtual acoustic space stimuli, we show that these acoustical changes are not by themselves responsible for the emergence of SC map topography. Presenting stimuli to infant ferrets through virtual adult ears did not improve the order in the representation of sound azimuth in the SC. But by using linear discriminant analysis to compare different response properties across age, we found that the SRFs of infant neurons nevertheless became more adult-like when stimuli were delivered through virtual adult ears. Hence, although the emergence of auditory topography is likely to depend on refinements in neural circuitry, maturation of the structure of the SRFs (particularly their spatial extent) can be largely accounted for by changes in the acoustics associated with growth of the head and ears.

Brief sounds evoke prolonged responses in anesthetized ferret auditory cortex.

Neurons in the auditory cortex of anesthetized animals are generally considered to generate phasic responses to simple stimuli such as tones or noise bursts. In this paper, we show that under ketamine/medetomidine anesthesia, neurons in ferret auditory cortex usually exhibit complex sustained responses. We presented 100-ms broad-band noise bursts at a range of interaural level differences (ILDs) and average binaural levels (ABLs), and used extracellular electrodes to monitor evoked activity over 700 ms poststimulus onset. We estimated the degree of randomness (noise) in the response functions of individual neurons over poststimulus time; we found that neural activity was significantly modulated by sound for up to ∼500 ms following stimulus offset. Pooling data from all neurons, we found that spiking activity carries significant information about stimulus identity over this same time period. However, information about ILD decayed much more quickly over time compared with information about ABL. In addition, ILD and ABL are coded independently by the neural population even though this is not the case at individual neurons. Though most neurons responded more strongly to ILDs corresponding to the opposite side of space, as a population, they were equally informative about both contra- and ipsilateral stimuli.

The mushroom body

What is a mushroom body? The mushroom body is a prominent and striking structure in the brain of several invertebrates, mainly arthropods. It is found in insects, scorpions, spiders, and even segmented worms. With its long stalk crowned with a cap of cell bodies, a GFP-labeled mushroom body certainly lives up to its name (Figure 1). The mushroom body is composed of small neurons known as Kenyon cells, named after Frederick Kenyon, who first applied the Golgi staining technique to the insect brain. The honey bee brain, for instance, contains roughly 175,000 neurons per mushroom body while the brain of the smaller fruit fly Drosophila melanogaster only possesses about 2,500. Kenyon cells thus make up 20% and 2%, respectively, of the total number of neurons in each insects brain. Kenyon cell bodies sit atop the calyx, a tangled zone of synapses representing the site of sensory input. Projecting away from the calyx is the stalk comprised of Kenyon cell axons carrying information away to the output lobes.

OpenStage: a low-cost motorized microscope stage with sub-micron positioning accuracy.

Recent progress in intracellular calcium sensors and other fluorophores has promoted the widespread adoption of functional optical imaging in the life sciences. Home-built multiphoton microscopes are easy to build, highly customizable, and cost effective. For many imaging applications a 3-axis motorized stage is critical, but commercially available motorization hardware (motorized translators, controller boxes, etc) are often very expensive. Furthermore, the firmware on commercial motor controllers cannot easily be altered and is not usually designed with a microscope stage in mind. Here we describe an open-source motorization solution that is simple to construct, yet far cheaper and more customizable than commercial offerings. The cost of the controller and motorization hardware are under

Specificity of binaural perceptual learning for amplitude modulated tones: A comparison of two training methods

1000. Hardware costs are kept low by replacing linear actuators with high quality stepper motors. Electronics are assembled from commonly available hobby components, which are easy to work with. Here we describe assembly of the system and quantify the positioning accuracy of all three axes. We obtain positioning repeatability of the order of in X/Y and in Z. A hand-held control-pad allows the user to direct stage motion precisely over a wide range of speeds ( to ), rapidly store and return to different locations, and execute “jumps” of a fixed size. In addition, the system can be controlled from a PC serial port. Our “OpenStage” controller is sufficiently flexible that it could be used to drive other devices, such as micro-manipulators, with minimal modifications.

Auditory Neuroscience: A Time for Coincidence?

The specificity of auditory perceptual learning has been taken as an indicator of the likely locus within the brain at which underlying neuronal changes occur. This study examined interaural level difference (ILD) discrimination learning with sinusoidally amplitude modulated (SAM) tones and whether training-induced threshold improvements generalize from one side of auditory space to the other and to an untrained carrier frequency. A novel, dual-staircase adaptive method was adopted that was designed to prevent participants from identifying the nature of the adaptive track. ILD thresholds obtained with this method were compared with a constant-stimulus technique using otherwise identical stimuli. Adaptive thresholds derived from psychometric functions were found to be biased compared to those obtained from reversals. Although adaptive and constant-stimulus procedures appeared to yield different temporal patterns of learning, no global differences were found between them in terms of training outcomes. These data show that ILD discrimination learning with SAM tones does generalize to an untrained carrier frequency but does not generalize across the midline. This implies that the neural substrate for binaural plasticity is found at a relatively high level of the auditory pathway where information is combined across frequency and where each side of auditory space is represented separately.

Two-Photon Imaging of Population Activity with Genetically Encoded Calcium Indicators in Living Flies

Mammals and birds appear to encode timing differences between the ears, a major cue for auditory localization, in fundamentally different ways. It now appears that results from different species can be accommodated within a single general framework.