Autophagy is activated in vivo during trimethyltin-induced apoptotic neurodegeneration: A study in the rat hippocampus

Trimethyltin (TMT) is an organotin compound known to produce significant and selective neuronal degeneration and reactive astrogliosis in the rodent central nervous system. Autophagy is the main cellular mechanism for degrading and recycling protein aggregates and damaged organelles, which in different stress conditions, such as starvation, generally improves cell survival. Autophagy is documented in several pathologic conditions, including neurodegenerative diseases. This study aimed to investigate the autophagy and apoptosis signaling pathways in hippocampal neurons of TMT-treated (Wistar) rats to explore molecular mechanisms involved in toxicant-induced neuronal injury. The microtubule-associated protein light chain (LC3, autophagosome marker) and sequestosome1 (SQSTM1/p62) (substrate of autophagy-mediated degradation) expressions were examined by Western blotting at different time points after intoxication. The results demonstrate that the LC3 II/I ratio significantly increased at 3 and 5 days, and that p62 levels significantly decreased at 7 and 14 days. Immunofluorescence images of LC3/neuronal nuclear antigen (NeuN) showed numerous strongly positive LC3 neurons throughout the hippocampus at 3 and 5 days. The terminal deoxynucleotidyltransferase dUTP nick end labeling (TUNEL) assay indicated an increase in apoptotic cells starting from 5 days after treatment. In order to clarify apoptotic pathway, immunofluorescence images of apoptosis-inducing factor (AIF)/NeuN did not show nuclear translocation of AIF in neurons. Increased expression of cleaved Caspase-3 was revealed at 5–14 days in all hippocampal regions by Western blotting and immunohistochemistry analyses. These data clearly demonstrate that TMT intoxication induces a marked increase in both autophagy and caspase-dependent apoptosis, and that autophagy occurring just before apoptosis could have a potential role in neuronal loss in this experimental model of neurodegeneration

GLI1 and AXIN2 Are Distinctive Markers of Human Calvarial Mesenchymal Stromal Cells in Nonsyndromic Craniosynostosis

All skeletal bones house osteogenic stem cell niches, in which mesenchymal stromal cells (MSC) provide progenitors for tissue growth and regeneration. They have been widely studied in long bones formed through endochondral ossification. Limited information is available on the composition of the osteogenic niche in flat bones (i.e., skull vault bones) that develop through direct membranous ossification. Craniosynostosis (CS) is a congenital craniofacial defect due to the excessive and premature ossification of skull vault sutures. This study aimed at analysing the expression of GLI1, AXIN2 and THY1 in the context of the human skull vault, using nonsyndromic forms of CS (NCS) as a model to test their functional implication in the aberrant osteogenic process. The expression of selected markers was studied in NCS patients' calvarial bone specimens, to assess the in vivo location of cells, and in MSC isolated thereof. The marker expression profile was analysed during in vitro osteogenic differentiation to validate the functional implication. Our results show that GLI1 and AXIN2 are expressed in periosteal and endosteal locations within the osteogenic niche of human calvarial bones. Their expression is higher in MSC isolated from calvarial bones than in those isolated from long bones and tends to decrease upon osteogenic commitment and differentiation. In particular, AXIN2 expression was lower in cells isolated from prematurely fused sutures than in those derived from patent sutures of NCS patients. This suggests that AXIN2 could reasonably represent a marker for the stem cell population that undergoes depletion during the premature ossification process occurring in CS

Lymphomyeloid organs of the Antarctic fish Trematomus nicolai and Chionodraco hamatus (Teleostei: Notothenioidea): a comparative histological study.

Lymphomyeloid organs of two common spe- cies of Antarctic fish, Trematomus nicolai and Chiono- draco hamatus, were studied with the aim of analysing some morphological aspects of these organs in relation to adaptation to low environmental temperature. The thymuses of T. nicolai and C. hamatus were flattened, incompletely lobated, with numerous Hassall-like bod- ies, which were mainly located in the central part of the organ in C. hamatus. In T. nicolai, thymocytes, erythroid and reticular epithelial cells filled the organ. In C. hamatus, the thymocytes intermingled with reticular epithelial cells were often close to groups of melano- macrophages. In both species, the thymus did not show distinct compartmentalisation; however, the thymocytes had significantly different sizes in the outer and inner portions of the thymus. The head kidney of both species was completely filled by haematopoietic tissue, highly vascularised and mainly lymphopoietic in T. nicolai, while both erythropoietic and lymphopoietic in C. hamatus. The spleen appeared mainly erythropoietic in T. nicolai and mainly lymphopoietic in C. hamatus. Solitary melano-macrophages in T. nicolai were close to numerous small vascular ellipsoids where erythroid and lymphoid cells were intermingled without the formation of red and white pulp areas. In C. hamatus, large lym- phoid areas were organised around the capillaries. The possible adaptation of lymphoid organs to the low temperature of polar water is discussed

Cytology of lymphomyeloid head kidney of Antarctic fishes Trematomus bernacchii (Nototheniidae) and Chionodraco hamatus (Channicthyidae)

Species that live in extreme conditions have specially adapted physiology and tissue/organ organisation. The adaptation of lymphoid organs to low temperatures in polar species could be an original field of study, indicating how the immune system works under extreme conditions. In fishes, the head kidney is a key organ for immunity and here the cytology of this organ is studied in two common Antarctic species: Trematomus bernacchii and Chionodraco hamatus. Ultrastructural analysis revealed heterogeneity of epithelial cells, with reticular cells, subcapsular- and perivascular-limiting cells. Differences in the size and morphology of epithelial cells were observed between the polar species and warm water species of fish. Intermingled with epithelial cell leucocytes, such as lymphocytes, thrombocytes and macrophages, had comparable morphology in both species, contrary to sharp differences observed in the morphology of erythrocytes and granulocytes. The functional adaptation of the head kidney to the low temperatures of polar water is discussed. \ua9 2002 Elsevier Science Ltd. All rights reserved

Adaptation of fish lymphomyeloid organs to polar water.

Lymphomyeloid organs of three common Antarctic fish species, Trematomus bernacchii, Trematomus nicolai and Chionodraco hamatus, were analysed. Contrary to species living in temperate sea water, the thymus of polar fishes were flattened, incompletely lobated and scarcely distinguishable by normal histology into cortical and medullary regions. Functional regionalisation, however, was suggested by differences in the sizes of thymocytes from the outer to the inner thymus zone. Another particularity was observed in the thymus of Trematomus species: next to lymphocytes, numerous erythroid cells circulated and differentiated in the parenchyma. Only two main types of epithelial cells could be found by cytological analysis: (i) limiting cells that surround the haematopoietic tissue and (ii) reticular cells that constitute the frame where the lymphoid and erythroid cells can proliferate and differentiate. The reticular cells could not be distinguished in cortical and medullary subtypes as observed in temperate-water fish. Numerous Hassall’s corpuscles, probably with a scavenging role, were also observed in the thymus. The head kidney housed haematopoietic tissue, lacked any excretory tubules, and had a huge blood supply, characteristic of polar fish species. It appeared mainly lymphopoietic in C. hamatus but contemporary erythropoietic and lymphopoietic in Trematomus species. The ultrastructural analysis revealed the presence of both reticular and limiting epithelial cells. Reticular epithelial cells (REC) characteristically showed numerous vesicles with a granular content and cell debris. Numerous lymphoblasts, lymphocytes and plasma cells were observed among the REC. Erythropoiesis occurred in all polar species analysed, but in C. hamatus the erythroblasts did not differentiate because they had a fast senescence. The spleen appeared mainly erythropoietic, with scarcely developed areas of white pulp, in Trematomus species; the erythropoiesis was scarcely evident in C. hamatus. Small vascular ellipsoids showed numerous melano-macrophages in Trematomus, while large haematopoietic areas were organised around the capillaries in C. hamatus. Utrastructural analysis revealed, in all species examined, two main types of epithelial cells: reticular, close to the ellipsoids, and limiting-subcapsular, which surround the organ. A large blood supply and extended capillary frame were also observed in polar species. The possible adaptation of lymphoid organs to the low temperatures of polar water is discussed

Lymphomyeloid organs of the Antarctic fish Trematomus nicolai and Chionodraco hamatus (Teleostei: Notothenioidea): A comparative histological study

Lymphomyeloid organs of two common species of Antarctic fish, Trematomus nicolai and Chionodraco hamatus, were studied with the aim of analysing some morphological aspects of these organs in relation to adaptation to low environmental temperature. The thymuses of T. nicolai and C. hamatus were flattened, incompletely lobated, with numerous Hassall-like bodies, which were mainly located in the central part of the organ in C. hamatus. In T. nicolai, thymocytes, erythroid and reticular epithelial cells filled the organ. In C. hamatus, the thymocytes intermingled with reticular epithelial cells were often close to groups of melano-macrophages. In both species, the thymus did not show distinct compartmentalisation; however, the thymocytes had significantly different sizes in the outer and inner portions of the thymus. The head kidney of both species was completely filled by haematopoietic tissue, highly vascularised and mainly lymphopoietic in T. nicolai, while both erythropoietic and lymphopoietic in C. hamatus. The spleen appeared mainly erythropoietic in T. nicolai and mainly lymphopoietic in C. hamatus. Solitary melano-macrophages in T. nicolai were close to numerous small vascular ellipsoids where erythroid and lymphoid cells were intermingled without the formation of red and white pulp areas. In C. hamatus, large lymphoid areas were organised around the capillaries. The possible adaptation of lymphoid organs to the low temperature of polar water is discussed

Adaptation of fish lymphomyeloid organs to polar water

Lmphomyeloid organs of three common Antarctic fish species, Trematomus bernacchii, Trematomus nicolai and Chionodraco hamatus, were analysed. Contrary to species living in temperate sea water, the thymus of polar fishes were flattened, incompletely lobated and scarcely distinguishable by normal histology into cortical and medullary regions. Functional regionalisation, however, was suggested by differences in the sizes of thymocytes from the outer to the inner thymus zone. Another particularity was observed in the thymus of Trematomus species: next to lymphocytes, numerous erythroid cells circulated and differentiated in the parenchyma. Only two main types of epithelial cells could be found by cytological analysis: (i) limiting cells that surround the haematopoietic tissue and (ii) reticular cells that constitute the frame where the lymphoid and erythroid cells can proliferate and differentiate. The reticular cells could not be distinguished in cortical and medullary subtypes as observed in temperate-water fish. Numerous Hassall's corpuscles, probably with a scavenging role, were also observed in the thymus. The head kidney housed haematopoietic tissue, lacked any excretory tubules, and had a huge blood supply, characteristic of polar fish species. It appeared mainly lymphopoietic in C. hamatus but contemporary erythropoietic and lymphopoietic in Trematomus species. The ultrastructural analysis revealed the presence of both reticular and limiting epithelial cells. Reticular epithelial cells (REC) characteristically showed numerous vesicles with a granular content and cell debris. Numerous lymphoblasts, lymphocytes and plasma cells were observed among the REC. Erythropoiesis occurred in all polar species analysed, but in C. hamatus the erythroblasts did not differentiate because they had a fast senescence. The spleen appeared mainly erythropoietic, with scarcely developed areas of white pulp, in Trematomus species; the erythropoiesis was scarcely evident in C. hamatus. Small vascular ellipsoids showed numerous melano-macrophages in Trematomus, while large haematopoietic areas were organised around the capillaries in C. hamatus. Utrastructural analysis revealed, in all species examined, two main types of epithelial cells: reticular, close to the ellipsoids, and limiting-subcapsular, which surround the organ. A large blood supply and extended capillary frame were also observed in polar species. The possible adaptation of lymphoid organs to the low temperatures of polar water is discussed. \ua9 2004 Taylor and Francis Ltd

Antigen-dependent T lymphocytes (TcR\u3b2+) are primarily differentiated in the thymus rather than in other lymphoid tissues in sea bass (Dicentrarchus labrax, L.)

All jawed vertebrates share lymphocyte receptors that allow the recognition of pathogens and the discrimination between self and non-self antigens. The T cell transmembrane receptor (TcR) has a central role in the maturation and function of T lymphocytes in vertebrates via an important role in positive selection of the variable region of TcR \u3b1\u3b2/\u3b3\u3b4 chains. In this study, the TcR\u3b2 transcript expression and TcR\u3b2+ cell distribution during the ontogeny of the immune system of sea bass (Dicentrarchus labrax, L.) were analysed. RT-PCR analysis of larvae during early development demonstrated that the \u3b2 chain transcript is expressed by 19 days post-fertilisation (p.f.). RNA probes specific for the \u3b2 chain were synthesised and used for in situ hybridisation experiments on 30 day p.f. to 180 day old juvenile larvae. A parallel immunohistochemical study was performed using the anti-T cell monoclonal antibody DLT15 developed in our laboratory [Scapigliati et al., Fish Shellfish Immunol 1996; 6:383-401]. The first thymus anlage was detectable at 32-33 days p.f. (Corresponding to about 27 days post-hatch). DLT15+ cells were detected at day 35 p.f. in the thymus whereas TcR\u3b2+ cells were recognisable at day 38 p.f. in the thymus and at day 41 p.f. in the gut. TcR\u3b2+ cells were observed in capillaries from 41 to 80 days p.f. At day 46 p.f., TcR\u3b2+ cells were identified in the head kidney and were detected in the spleen 4 days later. The present results demonstrate that TcR\u3b2+ cells can be differentiated first in the thymus and then in other organs/tissues, suggesting potential TcR\u3b2+ cell colonisation from the thymus to the middle gut. Once the epithelial architecture of the thymus is completed with the formation of the cortical-medullary border (around 70-75 days p.f.), DLT15+ cells or TcR\u3b2+ cells are confined mainly to the cortex and cortical-medullary border. In particular, a large influx of TcR\u3b2+ cells was observed at the cortical-medullary border from 72 to 90 days p.f., suggesting a role in positive selection for this thymic region during the ontogeny of the fish immune system. This study provides novel information about the primary differentiation and distribution of TcR\u3b2+ cells in sea bass larvae and juveniles. \ua9 2011 Elsevier Ltd