Annexin A2 (AnxA2) is a multifunctional calcium2+ (Ca2+) and phospholipid-binding protein that is expressed in a wide spectrum of cells, including those participating in the inflammatory response. In acute inflammation, the interaction of AnxA2 with actin and adherens junction VE-cadherins underlies its role in regulating vascular integrity. In addition, its contribution to endosomal membrane repair impacts several aspects of inflammatory regulation, including lysosome repair, which regulates inflammasome activation, and autophagosome biogenesis, which is essential for macroautophagy. On the other hand, AnxA2 may be co-opted to promote adhesion, entry, and propagation of bacteria or viruses into host cells. In the later stages of acute inflammation, AnxA2 contributes to the initiation of angiogenesis, which promotes tissue repair, but, when dysregulated, may also accompany chronic inflammation. AnxA2 is overexpressed in malignancies, such as breast cancer and glioblastoma, and likely contributes to cancer progression in the context of an inflammatory microenvironment. We conclude that annexin AnxA2 normally fulfills a spectrum of anti-inflammatory functions in the setting of both acute and chronic inflammation but may contribute to disease states in settings of disordered homeostasis.

Organotypic tissue cultures are highly promising for performing in vivo type studies in vitro. Currently, however, very limited survival times of only a few days for adult tissue often severely limit their application. Here, superhydrophilic nanostructured substrates with ideal material properties ensure tissue adhesion, essential for organotypic culture, while migration of single cells out of the tissue is hampered. Tuning substrate properties, for the first time, adult neuronal tissue could be cultured for 14 days with no indications of degeneration.

PurposeA population of corneal neurons in rats preferentially sense and monitor the hyperosmolar conditions of tears when the tears begin to evaporate during corneal dryness. The present study exploited this ability in an effort to estimate tear osmolarities by comparing the responses to corneal dryness to their responses to hyperosmolar stimuli.MethodsExtracellular recordings were performed from single neurons in the trigeminal ganglia innervating the corneas of rats. To determine the extent to which the corneal neurons' responses to drying of the cornea were induced via the activation by hyperosmolar stimuli, we assessed the responses to ocular instillation of 500 and 600 mOsm/L, and a graded series of hyperosmolar stimuli ranging from 350 to 1000 mOsm/L.ResultsThe magnitudes of the responses to drying of the cornea were matched almost exactly to those induced by the ocular instillation of the 600 mOsm/L stimuli but not the 500 mOsm/L solutions. The response magnitudes to a graded series of hyperosmolar solutions were nearly linear from the 350 to the 600 mOsm/L stimuli, but reached a plateau or declined slightly thereafter.ConclusionsOur results demonstrate that the tear osmolarity in rats could reach 600 to 1000 mOsm/L during ocular dryness. Furthermore, a spontaneous eye blink could be generated at a tear osmolarity of approximately 400 mOsm/L if the blink is solely determined by hyperosmolar tears, but ocular surface cooling also can become a major factor if hyperosmolar tears occurring during ocular dryness lower the threshold of activation of the neurons.

It is widely accepted that the mechanisms for transducing sensory information reside in the nerve terminals. Occasionally, however, studies have appeared demonstrating that similar mechanisms may exist in the axon to which these terminals are connected. We examined this issue in the cornea, where nerve terminals in the epithelial cell layers are easily accessible for debridement, leaving the underlying stromal (axonal) nerves undisturbed. In isoflurane-anesthetized rats, we recorded extracellularly from single trigeminal ganglion neurons innervating the cornea that are excited by ocular dryness and cooling: low-threshold (<2°C cooling) and high-threshold (>2°C) cold-sensitive plus dry-sensitive neurons playing possible roles in tearing and ocular pain. We found that the responses in both types of neurons to dryness, wetness, and menthol stimuli were effectively abolished by the debridement, indicating that their transduction mechanisms lie in the nerve terminals. However, some responses to the cold, heat, and hyperosmolar stimuli in low-threshold cold-sensitive plus dry-sensitive neurons still remained. Surprisingly, the responses to heat in approximately half of the neurons were augmented after the debridement. We were also able to evoke these residual responses and follow the trajectory of the stromal nerves, which we subsequently confirmed histologically. The residual responses always disappeared when the stromal nerves were cut at the limbus, suggesting that the additional transduction mechanisms for these sensory modalities originated most likely in stromal nerves. The functional significance of these residual and enhanced responses from stromal nerves may be related to the abnormal sensations observed in ocular disease. In addition to the traditional view that the sensory transduction mechanisms exist in the nerve terminals, we report here that the proximal axons (stromal nerves in the cornea from which these nerve terminals originate) may also be capable of transducing sensory information. We arrived at this conclusion by removing the epithelial cell layers of the cornea in which the nerve terminals reside but leaving the underlying stromal nerves undisturbed.

PurposePreviously we found two types of corneal neurons that we hypothesized to play an important role in tearing. One type is called low threshold–cold sensitive plus dry sensitive (LT-CS + DS), and the other is termed high threshold–cold sensitive plus dry sensitive (HT-CS + DS). The present study examined critical stimuli influencing the activity of these neurons to elucidate environmental factors that may trigger this ocular reflex.MethodsSingle corneal neurons were extracellularly recorded from the trigeminal ganglia in response to ocular stimuli that mimic environmental conditions one encounters in daily life. They included an ocular desiccation and slight air currents and were presented while simultaneously monitoring the ocular surface temperatures (OST) in rats.ResultsThe results showed that the changes in steady state (SS) activity of the neurons closely followed the changes in SS OST: during the sustained ocular desiccation, neural firing displayed numerous small sudden increases in activities (“spiking”); these “spiking” activities of LT-CS + DS neurons were replicated by a minute air current that induced slight ocular surface cooling of approximately 0.2–0.1°C; and the responses of HT-CS + DS neurons showed an inconsistent relationship to the changes in SS OST or exhibited little evidence for “spiking” activities.ConclusionsThese results suggest that LT-CS + DS neurons play a role in the afferent trigger of tearing as we face the environment, exposing the cornea to prevailing air currents that produce a slight cooling of the ocular surface. By contrast, HT-CS + DS neurons may serve to protect the eyes from extreme dryness by eliciting nociception-evoked tearing when the OST or osmolarity of tears becomes injurious.