Consistency and Variation in Doublecortin and Ki67 Antigen Detection in the Brain Tissue of Different Mammals, including Humans.

Recently, a population of "immature" neurons generated prenatally, retaining immaturity for long periods and finally integrating in adult circuits has been described in the cerebral cortex. Moreover, comparative studies revealed differences in occurrence/rate of different forms of neurogenic plasticity across mammals, the "immature" neurons prevailing in gyrencephalic species. To extend experimentation from laboratory mice to large-brained mammals, including humans, it is important to detect cell markers of neurogenic plasticity in brain tissues obtained from different procedures (e.g., post-mortem/intraoperative specimens vs. intracardiac perfusion). This variability overlaps with species-specific differences in antigen distribution or antibody species specificity, making it difficult for proper comparison. In this work, we detect the presence of doublecortin and Ki67 antigen, markers for neuronal immaturity and cell division, in six mammals characterized by widely different brain size. We tested seven commercial antibodies in four selected brain regions known to host immature neurons (paleocortex, neocortex) and newly born neurons (hippocampus, subventricular zone). In selected human brains, we confirmed the specificity of DCX antibody by performing co-staining with fluorescent probe for DCX mRNA. Our results indicate that, in spite of various types of fixations, most differences were due to the use of different antibodies and the existence of real interspecies variation.

Therapeutic induction of energy metabolism reduces neural tissue damage and increases microglia activation in severe spinal cord injury.

Neural tissue has high metabolic requirements. Following spinal cord injury (SCI), the damaged tissue suffers from a severe metabolic impairment, which aggravates axonal degeneration and neuronal loss. Impaired cellular energetic, tricarboxylic acid (TCA) cycle and oxidative phosphorylation metabolism in neuronal cells has been demonstrated to be a major cause of neural tissue death and regeneration failure following SCI. Therefore, rewiring the spinal cord cell metabolism may be an innovative therapeutic strategy for the treatment of SCI. In this study, we evaluated the therapeutic effect of the recovery of oxidative metabolism in a mouse model of severe contusive SCI. Oral administration of TCA cycle intermediates, co-factors, essential amino acids, and branched-chain amino acids was started 3 days post-injury and continued until the end of the experimental procedures. Metabolomic, immunohistological, and biochemical analyses were performed on the injured spinal cord sections. Administration of metabolic precursors enhanced spinal cord oxidative metabolism. In line with this metabolic shift, we observed the activation of the mTORC1 anabolic pathway, the increase in mitochondrial mass, and ROS defense which effectively prevented the injury-induced neural cell apoptosis in treated animals. Consistently, we found more choline acetyltransferase (ChAT)-expressing motor neurons and increased neurofilament-positive corticospinal axons in the spinal cord parenchyma of the treated mice. Interestingly, oral administration of the metabolic precursors increased the number of activated microglia expressing the CD206 marker suggestive of a pro-resolutive, M2-like phenotype. These molecular and histological modifications observed in treated animals ultimately led to a significant, although partial, improvement of the motor functions. Our data demonstrate that rewiring the cellular metabolism can represent an effective strategy to treat SCI.

Environmental Enrichment Induces Meningeal Niche Remodeling through TrkB-Mediated Signaling.

Neural precursors (NPs) present in the hippocampus can be modulated by several neurogenic stimuli, including environmental enrichment (EE) acting through BDNF-TrkB signaling. We have recently identified NPs in meninges; however, the meningeal niche response to pro-neurogenic stimuli has never been investigated. To this aim, we analyzed the effects of EE exposure on NP distribution in mouse brain meninges. Following neurogenic stimuli, although we did not detect modification of the meningeal cell number and proliferation, we observed an increased number of neural precursors in the meninges. A lineage tracing experiment suggested that EE-induced ß3-Tubulin+ immature neuronal cells present in the meninges originated, at least in part, from GLAST+ radial glia cells. To investigate the molecular mechanism responsible for meningeal reaction to EE exposure, we studied the BDNF-TrkB interaction. Treatment with ANA-12, a TrkB non-competitive inhibitor, abolished the EE-induced meningeal niche changes. Overall, these data showed, for the first time, that EE exposure induced meningeal niche remodeling through TrkB-mediated signaling. Fluoxetine treatment further confirmed the meningeal niche response, suggesting it may also respond to other pharmacological neurogenic stimuli. A better understanding of the neurogenic stimuli modulation for meninges may be useful to improve the effectiveness of neurodegenerative and neuropsychiatric treatments.

Protective effect of PACAP against ultraviolet B radiation-induced human corneal endothelial cell injury.

The human cornea, a sophisticated example of natural engineering, is composed in the innermost layer by endothelial cells maintaining stromal hydration and clarity. Different types of insults, including ultraviolet (UV) radiations, can lead to corneal opacity due to their degenerative and limited proliferative capability. In our previous studies, we have shown the protective effects of pituitary adenylate cyclase activating polypeptide (PACAP) in human corneal endothelial cells (HCECs), after growth factors deprivation. The aim of the present work has been to investigate the effect of this peptide on UV-B-induced HCECs injury. The results have shown that UV-B irradiations induced apoptotic cells death and consequently alteration in human corneal endothelial barrier. We found that PACAP treatment significantly increased viability, trans-endothelial electrical resistance and tight junctions expression of HCECs exposed to UV-B insult. In conclusion, data have suggested that this peptide could have protective effect to preserve the physiological state of human corneal endothelium exposed to UV-B damage.

Evaluation of the dentoskeletal characteristics of an Egyptian mummy with three-dimensional computer analysis.

BACKGROUND: Since the introduction of Roentgen rays in medical diagnostics, mummies have been subjected to radiographic and cephalometric studies. These have, among others, the advantage of providing details that are not directly visible for inspection without the need to tamper with the relics. The acquisition of three-dimensional imaging techniques has also extended the possibility of noninvasive investigation, so that many famous mummies, such as those of Tutankhamun and Ramses II, underwent three-dimensional computed tomography (CT). METHODS: Computed tomography scan of Egyptian mummy of a 20- to 30-year-old woman found in Fayum and dating from the second century B.C. has been performed. DICOM data of the CT scan have been processed by means of a software for three-dimensional CT imaging processing. The purpose of this report was to present the somatic and skeletal characteristics of the mummy. RESULTS: Thanks to the image processing, a "virtual reconstruction" of the original facial features of the mummy has been obtained, and a reliable cephalometric tracing could be performed. The data derived from cephalometric tracings were similar to those published on other studies on a group of Egyptian mummies and on a sample of Iowa adult males. CONCLUSIONS: In our opinion, three-dimensional image processing of CT scan is useful to perform noninvasive morphologic investigations on archeological find, to allow virtual correction of postmortem artifact and to perform reliable cephalometric tracings.