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The distribution and morphological characterization of nicotinamide adenine dinucleotide phosphate-diaphorase (ND)-positive cells and fibers in the tench central nervous system was mapped by using a direct histochemical method. This enzyme was observed in specific cell populations throughout all main divisions of the tench brain.In the telencephalon, we found strongly labeled olfactory fibers, as well as positive cells and fibers in the area ventralis of the telencephalic lobes. Positive staining was observed in the following diencephalic nuclei: nucleus preopticus magnocellularis pars magnocellularis, nucleus recessus lateralis, nucleus recessus posterioris, nucleus posterior tuberis, and nucleus diffusus torus lateralis, as well as small cells with a diffuse distribution surrounding the diencephalic ventricle. In the mesencephalon, heavily stained ND-positive neurons were observed in the nucleus fasciculi longitudinalis medialis, nucleus nervi oculomotorius, and nucleus nervi trochlearis. In the hindbrain the most evident staining was observed as large neurons located in the nuclei of the cranial nerves, scattered positive cells located between the negative fibers of the cranial nerves, and in the nucleus fasciculi solitari. Finally, in the spinal cord, ND-positive cells and fibers were mainly located in the ventral horn.This distribution of ND labeling in the brain of the tench is significantly different from previous data on ND activity in the brain of terrestrial vertebrates and does not correlate with the presence and distribution patterns of several neurotransmitters and neuroactive substances in the teleost brain. o 199.5 Wiley-Liss, Inc.

The hedgehog, a macrosomatic insectivore with an extraordinary development of the olfactory structures, has a crucial value for any phylogenetic or comparative study in mammals. The distribution pattern and morphology of NADPH-diaphorase-active and calbindin D-28k-immunoreactive neurons were studied in the main and accessory olfactory bulbs of the hedgehog. NADPH-diaphorase (ND) staining was carried out by a direct histochemical method, and the calbindin D-28k (CaBP) immunoreaction by using a monoclonal antibody and the avidin-biotin-immunoperoxidase method. The possible coexistence of both markers was determined by sequential histochemical-immunohistochemical double labeling of the same sections. Specific neuronal populations were positive for both ND and CaBP markers. No cell colocalized both stains in the hedgehog olfactory bulb. A subpopulation of olfactory fibers, and a subpopulation of olfactory glomeruli, located on the medial side, were positive for ND. Surrounding both the ND-positive and ND-negative glomeruli, there were ND- and CaBP-positive periglomerular cells, the latter group being much more abundant. A subpopulation of superficial short-axon cells was CaBP positive but, contrary to what is observed in rodents, this neuronal type was always ND negative. In addition, three neuronal types were observed in the GL-EPL border after CaBP immunostaining. These neuronal types have not been previously described either in the hedgehog or in the rodent olfactory bulb. Horizontal cells and vertical cells of Cajal were also observed after both ND and CaBP labeling. Distinct groups of ND- and CaBP-positive cells, differing in size, shape, dendritic branching pattern, and staining intensity, were distinguished in the granule cell layer and in the white matter. The large and medium-sized cells were identified as a very heterogeneous population of deep short-axon cells, whereas a subpopulation of granule cells was ND positive. The accessory olfactory bulb showed ND staining in all vomeronasal fibers and glomeruli, and in subpopulations of periglomerular cells, granule cells, and deep short-axon cells. The CaBP immunolabeling was more restricted and located in subpopulations of periglomerular cells and in deep short-axon cells. These results indicate different and more complex ND and CaBP staining patterns in the hedgehog olfactory bulb than those previously described in rodents, including the presence of specific, chemically and morphologically defined new neuronal types.

BackgroundGraft-versus-host disease (GvHD) remains the major obstacle to successful allogeneic hematopoietic stem cell transplantation, despite of the immunosuppressive regimens administered to control T cell alloreactivity. PI3K/AKT/mTOR pathway is crucial in T cell activation and function and, therefore, represents an attractive therapeutic target to prevent GvHD development. Recently, numerous PI3K inhibitors have been developed for cancer therapy. However, few studies have explored their immunosuppressive effect.MethodsThe effects of a selective PI3K inhibitor (BKM120) and a dual PI3K/mTOR inhibitor (BEZ235) on human T cell proliferation, expression of activation-related molecules, and phosphorylation of PI3K/AKT/mTOR pathway proteins were analyzed. Besides, the ability of BEZ235 to prevent GvHD development in mice was evaluated.ResultsSimultaneous inhibition of PI3K and mTOR was efficient at lower concentrations than PI3K specific targeting. Importantly, BEZ235 prevented naïve T cell activation and induced tolerance of alloreactive T cells, while maintaining an adequate response against cytomegalovirus, more efficiently than BKM120. Finally, BEZ235 treatment significantly improved the survival and decreased the GvHD development in mice.ConclusionsThese results support the use of PI3K inhibitors to control T cell responses and show the potential utility of the dual PI3K/mTOR inhibitor BEZ235 in GvHD prophylaxis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13045-016-0343-5) contains supplementary material, which is available to authorized users.