Impaired DNase1-mediated degradation of neutrophil extracellular traps is associated with acute thrombotic microangiopathies.

BACKGROUND: Acute thrombotic microangiopathies (TMAs) are characterized by excessive microvascular thrombosis and are associated with markers of neutrophil extracellular traps (NETs) in plasma. NETs are composed of DNA fibers and promote thrombus formation through the activation of platelets and clotting factors. OBJECTIVE: The efficient removal of NETs may be required to prevent excessive thrombosis such as in TMAs. To test this hypothesis, we investigated whether TMAs are associated with a defect in the degradation of NETs. METHODS AND RESULTS: We show that NETs generated in vitro were efficiently degraded by plasma from healthy donors. However, NETs remained stable after exposure to plasma from TMA patients. The inability to degrade NETs was linked to a reduced DNase activity in TMA plasma. Plasma DNase1 was required for efficient NET degradation and TMA plasma showed decreased levels of this enzyme. Supplementation of TMA plasma with recombinant human DNase1 restored NET-degradation activity. CONCLUSIONS: Our data indicate that DNase1-mediated degradation of NETs is impaired in patients with TMAs. The role of plasma DNases in thrombosis is, as of yet, poorly understood. Reduced plasma DNase1 activity may cause the persistence of pro-thrombotic NETs and thus promote microvascular thrombosis in TMA patients.

The muscarinic acetylcholine antagonist scopolamine impairs short-distance homing pigeon navigation.

The present study employed intramuscular (i.m.) injections of the acetylcholine (ACh) receptor antagonist scopolamine hydrobromide (0.10 mg/kg) to investigate the possible involvement of ACh in naturally occurring spatial navigation in homing pigeons (Columba livia). Control pigeons receiving injections of saline or scopolamine methylbromide, an ACh antagonist that does not cross the blood-brain barrier, were oriented in a homeward direction when released from a location 8 km from home. In contrast, pigeons injected with scopolamine hydrobromide (0.10 mg/kg, i.m.) were less well oriented and took more time to return home from the same location. These results suggest that homing pigeon navigation is regulated, in part, by central cholinergic mechanisms.

Evidence for muscarinic acetylcholine receptor subtypes in the pigeon telencephalon.

At least five subtypes of muscarinic acetylcholine receptors are expressed in various mammalian tissue preparations. The following experiment, through the use of direct binding assays (using tritiated quinuclidinyl benzilate), competitive binding assays (using tritiated quinuclidinyl benzilate and unlabeled pirenzepine or AF-DX 116), and autoradiographic techniques, examined whether two of these five putative muscarinic acetylcholine receptor subtypes can be found in avian brain. Accordingly, autoradiographic mapping of pirenzepine-sensitive (M1-like) and AF-DX 116-sensitive (M2-like) muscarinic acetylcholine receptor subtypes in the pigeon telencephalon was conducted. Although both ligands bound throughout the brain, most telencephalic regions, including the archistriatum, the neostriatum, and basal ganglia structures like lobus paraolfactorius, nucleus accumbens, and paleostriatum, showed a higher density of M1-like sites. The exception to this finding was the nucleus basalis which appeared as a region where M2-like sites predominated. Moreover, the telencephalic region with the largest ratio of M1-like to M2-like sites was the lateral portion of the parahippocampus; a characteristic shared with the mammalian dentate gyrus. The findings reported here are generally consistent with previous reports of mammalian M1/M2 receptor distributions.