Faculty Bibliography

Oculodentodigital dysplasia (ODDD) is a rare developmental disease resulting from germline mutations in the GJA1 gene that encodes the gap junction protein connexin43 (Cx43). In addition to the classical ODDD symptoms that affect the eyes, teeth, bone and digits, in some cases ODDD patients have reported bladder impairments. Thus, we chose to characterize the bladder in mutant mouse models of ODDD that harbor two distinct Cx43 mutations, G60S and I130T. Histological assessment revealed no difference in bladder detrusor wall thickness in mutant compared to littermate control mice. The overall localization of Cx43 in the lamina propria and detrusor also appeared to be similar in the bladders of mutant mice with the exception that the G60S mice had more instances of intracellular Cx43. However, both mutant mouse lines exhibited a significant reduction in the phosphorylated P1 and P2 isoforms of Cx43, while only the I130T mice exhibited a reduction in total Cx43 levels. Interestingly, Cx26 levels and distribution were not altered in mutant mice as it was localized to intracellular compartments and restricted to the basal cell layers of the urothelium. Our studies suggest that these two distinct genetically modified mouse models of ODDD probably mimic patients who lack bladder defects or other factors, such as aging or co-morbidities, are necessary to reveal a bladder phenotype.

Although there have been many advancements in cancer research, much is still unknown about the heterogeneous tumor microenvironment. Diffusion-weighted MRI has proven to be a viable and versatile microstructural probe. Diffusion-weighted sequences specifically sensitive to intravoxel incoherent motion (IVIM) have seen a recent resurgence of interest as they promise to provide a valuable window on the vascular microenvironment. To understand, test, and optimize IVIM-sensitive approaches, a complex flow phantom was constructed to mimic certain characteristics of the tumor microenvironment such as tortuous microvasculature, heterogeneous vascular permeability, and interstitial fluid pressure buildup. Results using this phantom on a clinical scanner platform confirmed IVIM sensitivity to microscopic flow effects. Biexponential fitting of signal decay curves enabled quantitative extraction of perfusion fraction, IVIM-related pseudodiffusivity, and tissue diffusivity. Parametric maps were also generated, illustrating the potential utility of IVIM-sensitive imaging in clinical settings. The flow phantom proved to be an effective test-bed for validating and optimizing the IVIM-MRI technique to provide surrogate markers for microvascular properties. Magn Reson Med, 2011. (c) 2011 Wiley Periodicals, Inc

Purpose:To assess the reproducibility and the distribution of intravoxel incoherent motion (IVIM) and diffusion-tensor (DT) imaging parameters in healthy renal cortex and medulla at baseline and after hydration or furosemide challenges.Materials and Methods:Using an institutional review board-approved HIPAA-compliant protocol with written informed consent, IVIM and DT imaging were performed at 3 T in 10 volunteers before and after water loading or furosemide administration. IVIM (apparent diffusion coefficient [ADC], tissue diffusivity [D(t)], perfusion fraction [f(p)], pseudodiffusivity [D(p)]) and DT (mean diffusivity [MD], fractional anisotropy [FA], eigenvalues [lambda(i)]) imaging parameters and urine output from serial bladder volumes were calculated. (a) Reproducibility was quantified with coefficient of variation, intraclass correlation coefficient, and Bland-Altman limits of agreement; (b) contrast and challenge response were quantified with analysis of variance; and (c) Pearson correlations were quantified with urine output.Results:Good reproducibility was found for ADC, D(t), MD, FA, and lambda(i) (average coefficient of variation, 3.7% [cortex] and 5.0% [medulla]), and moderate reproducibility was found for D(p), f(p), and f(p) . D(p) (average coefficient of variation, 18.7% [cortex] and 25.9% [medulla]). Baseline cortical diffusivities significantly exceeded medullary values except D(p), for which medullary values significantly exceeded cortical values, and lambda(1,) which showed no contrast. ADC, D(t), MD, and lambda(i) increased significantly for both challenges. Medullary diffusivity increases were dominated by transverse diffusion (1.72 +/- 0.09 [baseline] to 1.79 +/- 0.10 [hydration] mum(2)/msec, P = .0059; or 1.86 +/- 0.07 [furosemide] mum(2)/msec, P = .0094). Urine output correlated with cortical ADC with furosemide (r = 0.7, P = .034) and with medullary lambda(1) (r = 0.83, P = .0418), lambda(2) (r = 0.85, P = .0301), and MD (r = 0.82, P = .045) with hydration.Conclusion:Diffusion MR metrics are sensitive to flow changes in kidney induced by diuretic challenges. The results of this study suggest that vascular flow, tubular dilation, water reabsorption, and intratubular flow all play important roles in diffusion-weighted imaging contrast.(c) RSNA, 2012.

OBJECTIVE: The objective of this study was to review a single-institution contemporary experience with extracranial aneurysms of the carotid artery. METHODS: A retrospective review was conducted of patients evaluated for an aneurysm of the extracranial carotid artery from 2005 to 2010. Demographics, presentation, and operative management were reviewed. The mean follow-up was 22 months (range, 1-58 months). RESULTS: Over the study period, 16 aneurysms and pseudoaneurysms were identified in 15 patients. Of these, 14 aneurysms (in six men and eight women with mean age of 63 years) underwent surgical repair. The mean aneurysm size was 2.45 cm (range, 0.8-5 cm). One aneurysm (7.1%) was associated with neurologic symptoms, and 13 were asymptomatic. The underlying etiology was trauma in five (35.7%), prior carotid endarterectomy in five (35.7%), and degenerative atherosclerosis in four (28.6%). Aneurysms were isolated to the common carotid artery in six (42.9%), internal carotid artery in five (35.7%), and carotid bifurcation in three (21.4%). Five patients underwent aneurysmectomy with primary repair; seven underwent repair with an interposition graft, one required an innominate to common carotid artery bypass, and one patient had a plication and patch angioplasty. No mortalities or neurologic events were documented within 30 days. One patient had transient cranial nerve palsy. One patient required reintervention at 4 months for stenosis of the bypass graft, and one patient died at 10 months from an unrelated condition. There were no neurologic events on follow-up. CONCLUSIONS: Carotid artery aneurysms and pseudoaneurysms are uncommon and usually asymptomatic. Prior trauma and carotid surgery were common etiologies. The location of the aneurysms was equally distributed between the internal and common carotid arteries. Surgical repair was safe and effective with no significant morbidity or mortality and good midterm stroke prevention.

The dentate gyrus is one of two main areas of the mammalian brain where neurons are born throughout adulthood, a phenomenon called postnatal neurogenesis. Most of the neurons that are generated are granule cells (GCs), the major principal cell type in the dentate gyrus. Some adult-born granule cells develop in ectopic locations, such as the dentate hilus. The generation of hilar ectopic granule cells (HEGCs) is greatly increased in several animal models of epilepsy and has also been demonstrated in surgical specimens from patients with intractable temporal lobe epilepsy (TLE). Herein we review the results of our quantitative neuroanatomic analysis of HEGCs that were filled with Neurobiotin following electrophysiologic characterization in hippocampal slices. The data suggest that two types of HEGCs exist, based on a proximal or distal location of the cell body relative to the granule cell layer, and based on the location of most of the dendrites, in the molecular layer or hilus. Three-dimensional reconstruction revealed that the dendrites of distal HEGCs can extend along the transverse and longitudinal axis of the hippocampus. Analysis of axons demonstrated that HEGCs have projections that contribute to the normal mossy fiber innervation of CA3 as well as the abnormal sprouted fibers in the inner molecular layer of epileptic rodents (mossy fiber sprouting). These data support the idea that HEGCs could function as a "hub" cell in the dentate gyrus and play a critical role in network excitability.

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.

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.

The usefulness of genetically encoded probes for optical monitoring of neuronal activity and brain circuits would be greatly advanced by the generation of multiple indicators with non-overlapping color spectra. Most existing indicators are derived from or spectrally convergent on GFP. We generated a bright, red, pH-sensitive fluorescent protein, pHTomato, that can be used in parallel with green probes to monitor neuronal activity. SypHTomato, made by fusing pHTomato to the vesicular membrane protein synaptophysin, reported activity-dependent exocytosis as efficiently as green reporters. When expressed with the GFP-based indicator GCaMP3 in the same neuron, sypHTomato enabled concomitant imaging of transmitter release and presynaptic Ca(2+) transients at single nerve terminals. Expressing sypHTomato and GCaMP3 in separate cells enabled the simultaneous determination of presynaptic vesicular turnover and postsynaptic sub- and supra-threshold responses from a connected pair of neurons. With these new tools, we observed a close size matching between pre- and postsynaptic compartments, as well as interesting target cell-dependent regulation of presynaptic vesicle pools. Lastly, by coupling expression of pHTomato- and GFP-based probes with distinct variants of channelrhodopsin, we provided proof-of-principle for an all-optical approach to multiplex control and tracking of distinct circuit pathways.

Activity-dependent gene expression triggered by Ca(2+) entry into neurons is critical for learning and memory, but whether specific sources of Ca(2+) act distinctly or merely supply Ca(2+) to a common pool remains uncertain. Here, we report that both signaling modes coexist and pertain to Ca(V)1 and Ca(V)2 channels, respectively, coupling membrane depolarization to CREB phosphorylation and gene expression. Ca(V)1 channels are advantaged in their voltage-dependent gating and use nanodomain Ca(2+) to drive local CaMKII aggregation and trigger communication with the nucleus. In contrast, Ca(V)2 channels must elevate [Ca(2+)](i) microns away and promote CaMKII aggregation at Ca(V)1 channels. Consequently, Ca(V)2 channels are approximately 10-fold less effective in signaling to the nucleus than are Ca(V)1 channels for the same bulk [Ca(2+)](i) increase. Furthermore, Ca(V)2-mediated Ca(2+) rises are preferentially curbed by uptake into the endoplasmic reticulum and mitochondria. This source-biased buffering limits the spatial spread of Ca(2+), further attenuating Ca(V)2-mediated gene expression.