Faculty Bibliography

Agrin, an extracellular matrix protein belonging to the heterogeneous family of heparan sulfate proteoglycans (HSPGs), is expressed by cells of the hematopoietic system but its role in leukocyte biology is not yet clear. Here we demonstrate that agrin has a crucial, nonredundant role in myeloid cell development and functions. We have identified lineage-specific alterations that affect maturation, survival and properties of agrin-deficient monocytic cells, and occur at stages later than stem cell precursors. Our data indicate that the cell-autonomous signals delivered by agrin are sensed by macrophages through the alpha-DC (DG) receptor and lead to the activation of signaling pathways resulting in rearrangements of the actin cytoskeleton during the phagocytic synapse formation and phosphorylation of extracellular signal-regulated kinases (Erk 1/2). Altogether, these data identify agrin as a novel player of innate immunity.

Connexin-43 (Cx43), a gap junction protein involved in control of cell proliferation, differentiation and migration, has been suggested to have a role in hematopoiesis. Cx43 is highly expressed in osteoblasts and osteogenic progenitors (OB/P). To elucidate the biologic function of Cx43 in the hematopoietic microenvironment (HM) and its influence in hematopoietic stem cell (HSC) activity, we studied the hematopoietic function in an in vivo model of constitutive deficiency of Cx43 in OB/P. The deficiency of Cx43 in OB/P cells does not impair the steady state hematopoiesis, but disrupts the directional trafficking of HSC/progenitors (Ps) between the bone marrow (BM) and peripheral blood (PB). OB/P Cx43 is a crucial positive regulator of transstromal migration and homing of both HSCs and progenitors in an irradiated microenvironment. However, OB/P Cx43 deficiency in nonmyeloablated animals does not result in a homing defect but induces increased endosteal lodging and decreased mobilization of HSC/Ps associated with proliferation and expansion of Cxcl12-secreting mesenchymal/osteolineage cells in the BM HM in vivo. Cx43 controls the cellular content of the BM osteogenic microenvironment and is required for homing of HSC/Ps in myeloablated animals.

The Laboratory of Translational Auditory Research (LTAR/NYUSM) is part of the Department of Otolaryngology at the New York University School of Medicine and has close ties to the New York University Cochlear Implant Center. LTAR investigators have expertise in multiple related disciplines including speech and hearing science, audiology, engineering, and physiology. The lines of research in the laboratory deal mostly with speech perception by hearing impaired listeners, and particularly those who use cochlear implants (CIs) or hearing aids (HAs). Although the laboratory's research interests are diverse, there are common threads that permeate and tie all of its work. In particular, a strong interest in translational research underlies even the most basic studies carried out in the laboratory. Another important element is the development of engineering and computational tools, which range from mathematical models of speech perception to software and hardware that bypass clinical speech processors and stimulate cochlear implants directly, to novel ways of analyzing clinical outcomes data. If the appropriate tool to conduct an important experiment does not exist, we may work to develop it, either in house or in collaboration with academic or industrial partners. Another notable characteristic of the laboratory is its interdisciplinary nature where, for example, an audiologist and an engineer might work closely to develop an approach that would not have been feasible if each had worked singly on the project. Similarly, investigators with expertise in hearing aids and cochlear implants might join forces to study how human listeners integrate information provided by a CI and a HA. The following pages provide a flavor of the diversity and the commonalities of our research interests.

Neuronal oscillations allow for temporal segmentation of neuronal spikes. Interdependent oscillators can integrate multiple layers of information. We examined phase-phase coupling of theta and gamma oscillators in the CA1 region of rat hippocampus during maze exploration and rapid eye movement sleep. Hippocampal theta waves were asymmetric, and estimation of the spatial position of the animal was improved by identifying the waveform-based phase of spiking, compared to traditional methods used for phase estimation. Using the waveform-based theta phase, three distinct gamma bands were identified: slow gamma(S) (gamma(S); 30-50 Hz), midfrequency gamma(M) (gamma(M); 50-90 Hz), and fast gamma(F) (gamma(F); 90-150 Hz or epsilon band). The amplitude of each sub-band was modulated by the theta phase. In addition, we found reliable phase-phase coupling between theta and both gamma(S) and gamma(M) but not gamma(F) oscillators. We suggest that cross-frequency phase coupling can support multiple time-scale control of neuronal spikes within and across structures.

The CA3 and CA1 pyramidal neurons are the major principal cell types of the hippocampus proper. The strongly recurrent collateral system of CA3 cells and the largely parallel-organized CA1 neurons suggest that these regions perform distinct computations. However, a comprehensive comparison between CA1 and CA3 pyramidal cells in terms of firing properties, network dynamics, and behavioral correlations is sparse in the intact animal. We performed large-scale recordings in the dorsal hippocampus of rats to quantify the similarities and differences between CA1 (n > 3,600) and CA3 (n > 2,200) pyramidal cells during sleep and exploration in multiple environments. CA1 and CA3 neurons differed significantly in firing rates, spike burst propensity, spike entrainment by the theta rhythm, and other aspects of spiking dynamics in a brain state-dependent manner. A smaller proportion of CA3 than CA1 cells displayed prominent place fields, but place fields of CA3 neurons were more compact, more stable, and carried more spatial information per spike than those of CA1 pyramidal cells. Several other features of the two cell types were specific to the testing environment. CA3 neurons showed less pronounced phase precession and a weaker position versus spike-phase relationship than CA1 cells. Our findings suggest that these distinct activity dynamics of CA1 and CA3 pyramidal cells support their distinct computational roles. (c) 2012 Wiley Periodicals, Inc.

Cell type-specific expression of optogenetic molecules allows temporally precise manipulation of targeted neuronal activity. Here we present a toolbox of four knock-in mouse lines engineered for strong, Cre-dependent expression of channelrhodopsins ChR2-tdTomato and ChR2-EYFP, halorhodopsin eNpHR3.0 and archaerhodopsin Arch-ER2. All four transgenes mediated Cre-dependent, robust activation or silencing of cortical pyramidal neurons in vitro and in vivo upon light stimulation, with ChR2-EYFP and Arch-ER2 demonstrating light sensitivity approaching that of in utero or virally transduced neurons. We further show specific photoactivation of parvalbumin-positive interneurons in behaving ChR2-EYFP reporter mice. The robust, consistent and inducible nature of our ChR2 mice represents a significant advance over previous lines, and the Arch-ER2 and eNpHR3.0 mice are to our knowledge the first demonstration of successful conditional transgenic optogenetic silencing. When combined with the hundreds of available Cre driver lines, this optimized toolbox of reporter mice will enable widespread investigations of neural circuit function with unprecedented reliability and accuracy.

Neuronal control with high temporal precision is possible with optogenetics, yet currently-available methods do not enable to independently control multiple locations in the brains of freely-moving animals. Here, we describe a diode-probe system that allows real-time and location-specific control of neuronal activity at multiple sites. Manipulation of neuronal activity in arbitrary spatiotemporal patterns is achieved by means of an optoelectronic array, manufactured by attaching multiple diode-fiber assemblies to high-density silicon probes or wire tetrodes, and implanted into the brains of animals that are expressing light-responsive opsins. Each diode can be controlled separately, allowing localized light stimulation of neuronal activators and silencers in any temporal configuration and concurrent recording of the stimulated neurons. Because the only connections to the animals are via a highly flexible wire cable, unimpeded behavior is allowed for circuit monitoring and multi-site perturbations in the intact brain. The capacity of the system to generate unique neural activity patterns facilitates multi-site manipulation of neural circuits in a closed-loop manner and opens the door to addressing novel questions.

When a rat crosses the place field of a hippocampal pyramidal cell, this cell typically fires a series of spikes. Spike phases, measured with respect to theta oscillations of the local field potential, on average decrease as a function of the spatial distance traveled. This relation between phase and position of spikes might be a neural basis for encoding and is called phase precession. The degree of association between the circular phase variable and the linear spatial variable is commonly quantified through, however, a linear-linear correlation coefficient where the circular variable is converted to a linear variable by restricting the phase to an arbitrarily chosen range, which may bias the estimated correlation. Here we introduce a new measure to quantify circular-linear associations. This measure leads to a robust estimate of the slope and phase offset of the regression line, and it provides a correlation coefficient for circular-linear data that is a natural analog of Pearson's product-moment correlation coefficient for linear-linear data. Using surrogate data, we show that the new method outperforms the standard linear-linear approach with respect to estimates of the regression line and the correlation, and that the new method is less dependent on noise and sample size. We confirm these findings in a large data set of experimental recordings from hippocampal place cells and theta oscillations, and we discuss remaining problems that are relevant for the analysis and interpretation of phase precession. In summary, we provide a new method for the quantification of circular-linear associations.