Siddharth Srivastava

The Inflammasome Sensor, NLRP3, Regulates CNS Inflammation and Demyelination via Caspase-1 and Interleukin-18

Inflammation is increasingly recognized as an important contributor to a host of CNS disorders; however, its regulation in the brain is not well delineated. Nucleotide-binding domain, leucine-rich repeat, pyrin domain containing 3 (NLRP3) is a key component of the inflammasome complex, which also includes ASC (apoptotic speck-containing protein with a card) and procaspase-1. Inflammasome formation can be triggered by membrane P2X7R engagement leading to cleavage-induced maturation of caspase-1 and interleukin-1β (IL-1β)/IL-18. This work shows that expression of the Nlrp3 gene was increased >100-fold in a cuprizone-induced demyelination and neuroinflammation model. Mice lacking the Nlrp3 gene (Nlrp3−/−) exhibited delayed neuroinflammation, demyelination, and oligodendrocyte loss in this model. These mice also showed reduced demyelination in the experimental autoimmune encephalomyelitis model of neuroinflammation. This outcome is also observed for casp1−/− and IL-18−/− mice, whereas IL-1β−/− mice were indistinguishable from wild-type controls, indicating that Nlrp3-mediated function is through caspase-1 and IL-18. Additional analyses revealed that, unlike the IL-1β−/− mice, which have been previously shown to show delayed remyelination, Nlrp3−/− mice did not exhibit delayed remyelination. Interestingly, IL-18−/− mice showed enhanced remyelination, thus providing a possible compensatory mechanism for the lack of a remyelination defect in Nlrp3−/− mice. These results suggest that NLRP3 plays an important role in a model of multiple sclerosis by exacerbating CNS inflammation, and this is partly mediated by caspase-1 and IL-18. Additionally, the therapeutic inhibition of IL-18 might decrease demyelination but enhance remyelination, which has broad implications for demyelinating diseases.

A Distinct Representation of Three-Dimensional Shape in Macaque Anterior Intraparietal Area: Fast, Metric, and Coarse

Differences in the horizontal positions of retinal images—binocular disparity—provide important cues for three-dimensional object recognition and manipulation. We investigated the neural coding of three-dimensional shape defined by disparity in anterior intraparietal (AIP) area. Robust selectivity for disparity-defined slanted and curved surfaces was observed in a high proportion of AIP neurons, emerging at relatively short latencies. The large majority of AIP neurons preserved their three-dimensional shape preference over different positions in depth, a hallmark of higher-order disparity selectivity. Yet both stimulus type (concave–convex) and position in depth could be reliably decoded from the AIP responses. The neural coding of three-dimensional shape was based on first-order (slanted surfaces) and second-order (curved surfaces) disparity selectivity. Many AIP neurons tolerated the presence of disparity discontinuities in the stimulus, but the population of AIP neurons provided reliable information on the degree of curvedness of the stimulus. Finally, AIP neurons preserved their three-dimensional shape preference over different positions in the frontoparallel plane. Thus, AIP neurons extract or have access to three-dimensional object information defined by binocular disparity, consistent with previous functional magnetic resonance imaging data. Unlike the known representation of three-dimensional shape in inferior temporal cortex, the neural representation in AIP appears to emphasize object parameters required for the planning of grasping movements.

Coding of Shape and Position in Macaque Lateral Intraparietal Area

The analysis of object shape is critical for both object recognition and grasping. Areas in the intraparietal sulcus of the rhesus monkey are important for the visuomotor transformations underlying actions directed toward objects. The lateral intraparietal (LIP) area has strong anatomical connections with the anterior intraparietal area, which is known to control the shaping of the hand during grasping, and LIP neurons can respond selectively to simple two-dimensional shapes. Here we investigate the shape representation in area LIP of awake rhesus monkeys. Specifically, we determined to what extent LIP neurons are tuned to shape dimensions known to be relevant for grasping and assessed the invariance of their shape preferences with regard to changes in stimulus size and position in the receptive field. Most LIP neurons proved to be significantly tuned to multiple shape dimensions. The population of LIP neurons that were tested showed barely significant size invariance. Position invariance was present in a minority of the neurons tested. Many LIP neurons displayed spurious shape selectivity arising from accidental interactions between the stimulus and the receptive field. We observed pronounced differences in the receptive field profiles determined by presenting two different shapes. Almost all LIP neurons showed spatially selective saccadic activity, but the receptive field for saccades did not always correspond to the receptive field as determined using shapes. Our results demonstrate that a subpopulation of LIP neurons encodes stimulus shape. Furthermore, the shape representation in the dorsal visual stream appears to differ radically from the known representation of shape in the ventral visual stream.

Selectivity for three-dimensional contours and surfaces in the anterior intraparietal area

The macaque anterior intraparietal area (AIP) is crucial for visually guided grasping. AIP neurons respond during the visual presentation of real-world objects and encode the depth profile of disparity-defined curved surfaces. We investigated the neural representation of curved surfaces in AIP using a stimulus-reduction approach. The stimuli consisted of three-dimensional (3-D) shapes curved along the horizontal axis, the vertical axis, or both the horizontal and the vertical axes of the shape. The depth profile was defined solely by binocular disparity that varied along either the boundary or the surface of the shape or along both the boundary and the surface of the shape. The majority of AIP neurons were selective for curved boundaries along the horizontal or the vertical axis, and neural selectivity emerged at short latencies. Stimuli in which disparity varied only along the surface of the shape (with zero disparity on the boundaries) evoked selectivity in a smaller proportion of AIP neurons and at considerably longer latencies. AIP neurons were not selective for 3-D surfaces composed of anticorrelated disparities. Thus the neural selectivity for object depth profile in AIP is present when only the boundary is curved in depth, but not for disparity in anticorrelated stereograms.

Recombinant Human Interleukin-11 Treatment Enhances Collateral Vessel Growth After Femoral Artery Ligation

Objective—To investigate the role of recombinant human interleukin-11 (rhIL-11) on in vivo mobilization of CD34+/vascular endothelial growth factor receptor (VEGFR) 2+ mononuclear cells and collateral vessel remodeling in a mouse model of hindlimb ischemia. Methods and Results—We observed that treatment of Sv129 mice with continuous infusion of 200-&mgr;g/kg rhIL-11 per day led to in vivo mobilization of CD34+/VEGFR2+ cells that peaked at 72 hours. Sv129 mice pretreated with rhIL-11 for 72 hours before femoral artery ligation showed a 3-fold increase in plantar vessel perfusion, leading to faster blood flow recovery; and a 20-fold increase in circulating CD34+/VEGFR2+ cells after 8 days of rhIL-11 treatment. Histologically, experimental mice had a 3-fold increase in collateral vessel luminal diameter after 21 days of rhIL-11 treatment and a 4.4-fold influx of perivascular CD34+/VEGFR2+ cells after 8 days of therapy. Functionally, rhIL-11–treated mice showed better hindlimb appearance and use scores when compared with syngeneic mice treated with PBS under the same experimental conditions. Conclusion—These novel findings show that rhIL-11 promotes in vivo mobilization of CD34+/VEGFR2+ mononuclear cells, enhances collateral vessel growth, and increases recovery of perfusion after femoral artery ligation. Thus, rhIL-11 has a promising role for development as an adjunctive treatment of patients with peripheral vascular disease.

Characterization of Borrelia burgdorferi aggregates.

Lyme disease is caused by the tick-borne spirochete, Borrelia burgdorferi. It has been documented that B. burgdorferi form aggregates within ticks and during in vitro culture. However, Borrelia aggregates remain poorly characterized, and their functional significance is unknown. Here we have characterized Borrelia aggregates using microscopy and flow cytometry. Borrelia aggregation was temperature, pH, and growth phase dependent. Environmental conditions (high temperature, low pH, and high cell density) favorable for aggregation were similar to the conditions that increased the expression of B. burgdoferi genes, such as outer surface protein C (ospC), that are regulated by the RpoN/RpoS sigma factors. Experiments were conducted to determine if there is a relationship between aggregation and gene regulation through the RpoN/RpoS pathway. ospC Transcript levels were similar between aggregates and free cells. Moreover, no differences were observed in aggregate formation when null mutants of rpoS, rpoN, or ospC were compared to wild-type spirochetes. These results indicated that, despite the similar external signals that promoted aggregation and the RpoN/RpoS pathway, the two processes were not linked at the molecular level. The methods developed here to study B. burgdorferi aggregates will be useful for further studies on spirochete aggregates.

RECOMBINANT HUMAN INTERLEUKIN-11 TREATMENT IMPROVES REPERFUSION AND AUGMENTS COLLATERAL VESSEL GROWTH IN MOUSE MODEL OF SEVERE HINDLIMB ISCHEMIA

Results: Recombinant human interleukin-11 treatment resulted in 3-fold increase in plantar collateral vessel perfusion and 4-fold increase in collateral vessel growth (increase in luminal diameter) which correlated with a significant increase in hindlimb function measured by hindlimb appearanceand hindlimb use-score. There was a significant increase in both circulating and peri-collateral vessel CD34+/VEGFR2+ mononuclear cells and monocytes in rhIL-11-treated mice when compared with syngeneic mice treated with placebo under the same experimental condition.