Sponge Cells Research Articles

Analysis of the Effect of Locally Applied Bovine Collagen Sponge and Adipose-Derived Mesenchymal Stem Cells on Seroma Development in Rats Undergone Mastectomy and Axillary Dissection

ABSTRACTPurpose: Seroma is the most frequently seen complication after the mastectomy and axillary dissection. The aim of this study was to analyze the effects of locally applied bovine collagen sponge and adipose-derived mesenchymal stem cells on seroma development in rats that undergone mastectomy and axillary dissection. Materials and Methods: Wistar albino rats, were randomly divided into 3 groups (n = 10 per group). For the rats in Group 1, 1 ml 0.09% NaCl was implemented. 2 × 106/kg adipose-derived mesenchymal stem cell was implemented within 1 ml 0.09% NaCl for the rats in Group 2, and 3 cm2 bovine collagen sponge were locally applied for the rats in Group 3. Adhesion scores, histopathological examination, E-cadherin expression and tissue seroma volume were evaluated. Results: The seroma volume of Group 3 were significantly lower than those of Groups 2 and 1 (p < 0.001). General adhesion scores of Group 3 were significantly higher than those of Groups 1 and 2 (p < 0.05). Statistically significant increase was observed in Group 3 compared to Group 1 in terms of fibroblast, neovascularization and collagen density (p < 0.05). Conclusion: Local application of bovine collagen sponge and ADSCs in rats, which have undergone experimental mastectomy and axillary dissection, can be told to decrease the seroma formation and to increase the neovascularization and collagen deposition. This effect is more significant in bovine collagen sponge group.

Evolution and function of eukaryotic-like proteins from sponge symbionts.

Sponges (Porifera) are ancient metazoans that harbour diverse microorganisms, whose symbiotic interactions are essential for the host's health and function. Although symbiosis between bacteria and sponges are ubiquitous, the molecular mechanisms that control these associations are largely unknown. Recent (meta-) genomic analyses discovered an abundance of genes encoding for eukaryotic-like proteins (ELPs) in bacterial symbionts from different sponge species. ELPs belonging to the ankyrin repeat (AR) class from a bacterial symbiont of the sponge Cymbastela concentrica were subsequently found to modulate amoebal phagocytosis. This might be a molecular mechanism, by which symbionts can control their interaction with the sponge. In this study, we investigated the evolution and function of ELPs from other classes and from symbionts found in other sponges to better understand the importance of ELPs for bacteria-eukaryote interactions. Phylogenetic analyses showed that all of the nine ELPs investigated were most closely related to proteins found either in eukaryotes or in bacteria that can live in association with eukaryotes. ELPs were then recombinantly expressed in Escherichia coli and exposed to the amoeba Acanthamoeba castellanii, which is functionally analogous to phagocytic cells in sponges. Phagocytosis assays with E.coli containing three ELP classes (AR, TPR-SEL1 and NHL) showed a significantly higher percentage of amoeba containing bacteria and average number of intracellular bacteria per amoeba when compared to negative controls. The result that various classes of ELPs found in symbionts of different sponges can modulate phagocytosis indicates that they have a broader function in mediating bacteria-sponge interactions.

Spherulization as a process for the exudation of chemical cues by the encrusting sponge C. crambe.

Ecological interactions in the marine environment are now recognized to be partly held by chemical cues produced by marine organisms. In particular, sponges are sessile animals thought to rely on the bioactive substances they synthesize to ensure their development and defense. However, the mechanisms leading the sponges to use their specialized metabolites as chemical cues remain unknown. Here we report the constant release of bioactive polycyclic guanidinic alkaloids by the Mediterranean sponge Crambe crambe into the dissolved and the particulate phases using a targeted metabolomics study. These compounds were proven to be stored into already described specialized (spherulous) sponge cells and dispersed into the water column after release through the sponge exhaling channels (oscula), leading to a chemical shield surrounding the sponge. Low concentrations of these compounds were demonstrated to have teratogenic effects on embryos of a common sea squirt (ascidian). This mechanism of action called spherulization may therefore contribute to the ecological success of encrusting sponges that need to extend their substrate cover to expand.

The potential of azooxanthellate poriferan hosts to assess the fundamental and realized Symbiodinium niche: evaluating a novel method to initiate Symbiodinium associations

On coral reefs, Symbiodinium spp. are found in most cnidarian species, but reside in only a small number of sponge species. Of the sponges that do harbor Symbiodinium, most are found in the family Clionaidae, which represents a minor fraction of the poriferan diversity on a reef. Our goal was to determine whether Symbiodinium can be taken up by sponge hosts that do not typically harbor these algal symbionts, and then to follow the fate of any Symbiodinium that enter the intracellular space. We used the filter-feeding capacity of sponges to initiate intracellular interactions between sponge-specialist clade G Symbiodinium and six sponge species that do not associate with Symbiodinium. Using a pulse-chase experimental design, we determined that all of the species we examined captured Symbiodinium, and undamaged intracellular algae were found up to 1 h after inoculation. In a longer-term experiment, Symbiodinium populations in Amphimedon erina persisted in sponge cells for at least 5 d post-inoculation. While no evidence of digestion was detected, the population decreased exponentially after inoculation. We contrast these data with the characteristics of symbiont acquisition and establishment in Cliona varians, which normally harbors Symbiodinium. Explants from experimentally derived aposymbiotic sponges were placed in the field where they acquired Symbiodinium from ambient sources (i.e., we did not inoculate them as in the pulse-chase experiments). We began to detect Symbiodinium cells in C. varians after 12 d, and the algal population increased exponentially until densities approached those typically found in this host (after ~128 d). We discuss the implications of this work in light of growing interest in the evolution of specificity between hosts and symbionts, and the fundamental and realized niche of Symbiodinium.

Shift in aggregation, ROS generation, antioxidative defense, lysozyme and acetylcholinesterase activities in the cells of an Indian freshwater sponge exposed to washing soda (sodium carbonate)

Washing soda, chemically identified as anhydrous sodium carbonate, is a popular cleaning agent among the rural and urban populations of India which often contaminates the freshwater ponds and lakes, the natural habitat of sponge Eunapius carteri. Present investigation deals with estimation of cellular aggregation, generation of ROS and activities of antioxidant enzymes, lysozyme and acetylcholinesterase in the cells of E. carteri under the environmentally realistic concentrations of washing soda. Prolonged treatment of washing soda inhibited the degree of cellular aggregation. Experimental exposure of 8 and 16mg/l of sodium carbonate for 48h elevated the physiological level of reactive oxygen species (ROS) generation in the agranulocytes, semigranulocytes and granulocytes of E. carteri, whereas, treatment of 192h inhibited the ROS generation in three cellular morphotypes. Activities of superoxide dismutase, catalase and glutathione-S-transferase were recorded to be inhibited under prolonged exposure of washing soda. Washing soda mediated inhibition of ROS generation and depletion in the activities of antioxidant enzymes were indicative to an undesirable shift in cytotoxic status and antioxidative defense in E. carteri. Inhibition in the activity of lysozyme under the treatment of sodium carbonate was suggestive to a severe impairment of the innate immunological efficiency of E. carteri distributed in the washing soda contaminated habitat. Washing soda mediated inhibition in the activity of acetylcholinesterase indicated its neurotoxicity in E. carteri. Washing soda, a reported environmental contaminant, affected adversely the immunophysiological status of E. carteri with reference to cellular aggregation, oxidative stress, antioxidative defense, lysozyme and acetylcholinesterase activity.

A report of hailstorm damage to two species of freshwater sponge (Demospongiae: Haplosclerida: Spongillidae) populations of West Bengal, India

Selected freshwater bodies in Jagaddal of North 24 Parganas in West Bengal, India are inhabited by two species of sponge, Eunapius carteri (Bowerbank, 1863) and Spongilla alba (Carter, 1849) (Porifera: Demospongiae: Spongillidae). Most of these wetlands are perennial ponds without a history of aquaculture and toxin contamination. On 22 March 2014, the entire area of Jagaddal experienced an unprecedented hailstorm associated with a sharp decline of environmental temperature from 35 0C to 21 0C within 10–15 minutes. The hailstorm associated with torrential rain lasted for about 30 minutes. The natural habitat of the sponge was visited after six hours of the hailstorm in open day light conditions. During the field investigation, we recorded large-scale damage to the populations of E. carteri and S. alba. Macroscopic observation revealed that the fragmentation of body masses were also associated with cellular disintegration of the external surface. Sponge cells were experimentally dissociated from the sponge fragments and subjected to dye exclusion assay. A vital dye (trypan blue) exclusion assay of sponge fragments confirmed a high degree of mortality of the cells of E. carteri and S. alba. Hailstorm associated with the decline of environmental temperature down to 21 0C resulted in the mass destruction of these two species of sponges in their natural habitat. Any unprecedented and acute change in the climatic and hydrological parameter may lead to physiological adversity in the freshwater sponge. Subsequently, recovery from cold shock and mechanical stresses was overcome and a regeneration of sponge specimens was recorded within a period of around six months from the date of the hailstorm.

Seasonality and lunar periodicity in the sexual reproduction of the coral-killing sponge, Terpios hoshinota

Sexual reproduction of the cyanobacteriosponge, Terpios hoshinota, was studied at Lyudao (Green Island), Taiwan, from 2011 to 2013 through histological examinations, electron microscopy, and in situ observations of larval release. Histological examinations identified five reproductive structures: oocytes, sperm cysts, cell masses, early-stage embryos, and mature embryos. These reproductive structures were often observed in the same specimens, indicating that T. hoshinota is a hermaphroditic brooder. No cyanobacteria were seen in the gametes, and transmission of symbiotic cyanobacteria from parental tissues to larvae likely occurred during embryogenesis. The cell mass, a loose aggregate of numerous symbiotic cyanobacteria and maternal sponge cells, appeared to be eventually incorporated and constitutes the inner part of pre-hatching larvae, suggesting that the larval type is pseudoblastula instead of parenchymella as previously suggested. A clear lunar periodicity was seen in the reproductive cycle; larvae were released mostly around the full moon and occasionally around the new moon. Reproductive activity declined during months with low temperatures (January–April). The larvae were characterized by negative buoyancy and limited mobility, suggesting a larval dispersal distance on a scale of meters under calm weather. However, long-distance dispersal of larvae could still be possible through occasional strong currents during bad weather, such as typhoons, if the larvae survived during the dispersal period by accessing extra energy from the symbiotic cyanobacteria through their photosynthates and/or consuming the cyanobacteria as a source of nutrients. This study showed that T. hoshinota has high fecundity, with a monthly release of numerous larvae over a long reproductive season. This high fecundity, along with local larval dispersal and rapid post-settlement growth, enables rapid population expansion of T. hoshinota.

Spiculogenesis in the siliceous sponge Lubomirskia baicalensis studied with fluorescent staining.

Siliceous sponges are the most primitive multicellular animals whose skeleton consists of spicules - needle-like constructions from silicon dioxide surrounding organic axial filaments. Mechanisms of spicule formation have been intensively studied due to the high ecological importance of sponges and their interest to materials science. Light and electron microscopy are not appropriate enough to display the process from silicon-enriched cells to mature spicules because of composite structure of the sponge tissues. In this article, spiculogenesis in the siliceous sponge has been studied for the first time with the use of fluorescent microscopy. Fluorescent vital dye NBD-N2 was applied to stain growing siliceous structures in the sponge and primmorph cell system. The main stages of spicule growth in the fresh-water sponge Lubomirskia baicalensis (Pallas, 1773) were visualized: silicon accumulation in sclerocytes; formation of an organic filament protruding from the cell; further elongation of the filament and growth of the spicule in a spindle-like form with enlargement in the center; merger with new sclerocytes and formation of the mature spicule. Fluorescent microscopy combined with SEM allows us to overcome the virtual differentiation between intra- and extracellular mechanisms of spicule growth. The growing spicule can capture silicic acid from the extracellular space and merge with new silicon-enriched cells. Visualization of the growing spicules with the fluorescent dye allows us to monitor sponge viability in ecological or toxicological experiments and to apply genomic, proteomic and biochemical techniques.

A new tactile device using magneto-rheological sponge cells for medical applications: Experimental investigation

This work proposes a new tactile device that uses a single magneto-rheological (MR) sponge cell to realize the viscoelastic sensation of real organs. The proposed MR sponge cell can be usefully applied to medical fields to provide tactile information to surgeons who perform robotic surgery and doctors who receive training for practicing surgery with a computer-based simulator. In order to demonstrate the effectiveness of the proposed MR sponge cell, a 3-axis robot is designed and manufactured as an experimental apparatus. Using the robot, the relaxation times and repulsive forces of the MR sponge cell are measured and compared among three animal specimens: pork, pork rind, and pork heart. It is noted that the specimens represent the soft or/and hard tissues of human organs. The relaxation times are obtained from exponential equations based on the measurement data according to the properties of the viscoelastic material. The repulsive forces, normal force, tangential force, and bending moment are determined on the basis of comparison criteria of the beam bundle model (BBM). It is shown that the proposed MR sponge cell can effectively represent different feeling of the sensation of the specimens by applying different magnetic intensity to the cell domain.

Establishment of primmorphs from three Red Sea sponge species

Background Primmorphs are a special form of 3D-cell aggregates obtained from sponge cells. They can be used as biofermenters for the production of bioactive secondary metabolites. In the commercial development of sponge-derived drug leads, the production of primmorphs is one of the methods proposed to solve the supply problem. In addition, using primmorphs for the production of drugs can preserve the sponge population from extinction by producing enough quantities of the extracts and compounds that present in wild sponges. Objectives The presented work aimed to produce primmorphs of Red Sea sponges Hemimycale aff arabica , Stylissa carteri , and Crella (Yvesia) spinulata as long-term cultivation in vitro and identify the impact of different cell densities on their formation and growth. Results Microscopic studies suggested that primmorphs are formed through four stages: amorphous large cell floc within 1–3 h; small irregular cell aggregations in 1 day; large primary cell aggregations and round-shaped primmorphs after 3 days. Primmorphs of C. spinulata and S. carteri remained alive for 3–6 months. The primmorphs of H. arabica remained alive for 1 month. Long-term primmorph cultivation in vitro allows the creation of a controlled live model under experimental conditions. Conclusion This work may provide a solution to the ‘supply problem’ in the commercial development of sponge-derived drugs, as primmorphs can be used as biofermenters for bioactive secondary metabolite production. In addition, primmorphs can be used to study the morphogenesis of their sponges at different stages and transdifferentiation as well as the processes of spiculogenesis.

&amp;lt;i&amp;gt;In Vivo&amp;lt;/i&amp;gt; Estimation of Osteogenesis by Bone Marrow Cells in a Bi-Phasic Scaffold and in Each of Its Components

A bi-phasic scaffold consisting of a columnar formaldehyde-acetalized polyvinyl alcohol (PVF) sponge and a cylindrical porous hydroxyapatite (HA) with a hollow center was devised. Rat bone marrow cells (rBMCs) were seeded into the sponge placed in the hollow center of the cylindrical porous HA. The bi-phasic scaffold, a cylindrical porous HA and a PVF sponge separated from a bi-phasic scaffold after rBMC seeding, and a PVF sponge without rBMCs as a negative control, were implanted for 6 weeks into rat dorsal subcutaneous tissue. In each construct, bone formation was examined histologically and osteocalcin was measured immunochemically. Bone formation was observed in the bi-phasic scaffold and also in the cylindrical porous HA isolated from the bi-phasic scaffold. A significant difference in the quantity of osteocalcin was observed between the bi-phasic scaffold and the isolated cylindrical porous HA. No bone formation was found in the isolated PVF sponge. The bi-phasic scaffold as an outer layer of the scaffold seemed to inhibit the outflow of rBMCs from the PVF sponge. This type of bi-phasic scaffold may have two specific characteristics: Attachment of cells both in PVF sponge and cylindrical porous HA.

Sponges and their microbiomes in clean and polluted habitats

Sponges and their microbiomes in clean and polluted habitats

Biofouling of inlet pipes affects water quality in running seawater aquaria and compromises sponge cell proliferation.

Marine organism are often kept, cultured, and experimented on in running seawater aquaria. However, surprisingly little attention is given to the nutrient composition of the water flowing through these systems, which is generally assumed to equal in situ conditions, but may change due to the presence of biofouling organisms. Significantly lower bacterial abundances and higher inorganic nitrogen species (nitrate, nitrite, and ammonium) were measured in aquarium water when biofouling organisms were present within a 7-year old inlet pipe feeding a tropical reef running seawater aquaria system, compared with aquarium water fed by a new, biofouling-free inlet pipe. These water quality changes are indicative of the feeding activity and waste production of the suspension- and filter-feeding communities found in the old pipe, which included sponges, bivalves, barnacles, and ascidians. To illustrate the physiological consequences of these water quality changes on a model organism kept in the aquaria system, we investigated the influence of the presence and absence of the biofouling community on the functioning of the filter-feeding sponge Halisarca caerulea, by determining its choanocyte (filter cell) proliferation rates. We found a 34% increase in choanocyte proliferation rates following the replacement of the inlet pipe (i.e., removal of the biofouling community). This indicates that the physiological functioning of the sponge was compromised due to suboptimal food conditions within the aquarium resulting from the presence of the biofouling organisms in the inlet pipe. This study has implications for the husbandry and performance of experiments with marine organisms in running seawater aquaria systems. Inlet pipes should be checked regularly, and replaced if necessary, in order to avoid excessive biofouling and to approach in situ water quality.

Identification of okadaic acid binding protein 2 in reconstituted sponge cell clusters from Halichondria okadai and its contribution to the detoxification of okadaic acid

Okadaic acid (OA) and OA binding protein 2 (OABP2) were previously isolated from the marine sponge Halichondria okadai. Because the amino acid sequence of OABP2 is completely different from that of protein phosphatase 2A, a well-known target of OA, we have been investigating the production and function of OABP2. In the present study, we hypothesized that OABP2 plays a role in the detoxification of OA in H. okadai and that the OA concentrations are in proportional to the OABP2 concentrations in the sponge specimens. Based on the OA concentrations and the OABP2 concentrations in the sponge specimens collected in various places and in different seasons, however, we could not determine a positive correlation between OA and OABP2. We then attempted to determine distribution of OA and OABP2 in the sponge specimen. When the mixture of dissociated sponge cells and symbiotic species were separated with various pore-sized nylon meshes, most of the OA and OABP2 was detected from the same 0–10 μm fraction. Next, when sponge cell clusters were prepared from a mixture of dissociated sponge cells and symbiotic species in the presence of penicillin and streptomycin, we identified the 18S rDNA of H. okadai and the gene of OABP2 in the analysis of genomic DNA but could not detect OA by LC-MS/MS. We thus concluded that the sponge cells express OABP2, and that OA was not apparently present in the sponge cells but could be colocalized with OABP2 in the sponge cells at a concentration less than the limit of detection.

Morphological alteration, lysosomal membrane fragility and apoptosis of the cells of Indian freshwater sponge exposed to washing soda (sodium carbonate)

Washing soda is chemically known as sodium carbonate and is a component of laundry detergent. Domestic effluent, drain water and various anthropogenic activities have been identified as major routes of sodium carbonate contamination of the freshwater ecosystem. The freshwater sponge, Eunapius carteri, bears ecological and evolutionary significance and is considered as a bioresource in aquatic ecosystems. The present study involves estimation of morphological damage, lysosomal membrane integrity, activity of phosphatases and apoptosis in the cells of E. carteri under the environmentally realistic concentrations of washing soda. Exposure to washing soda resulted in severe morphological alterations and damages in cells of E. carteri. Fragility and destabilization of lysosomal membranes of E. carteri under the sublethal exposure was indicative to toxin induced physiological stress in sponge. Prolonged exposure to sodium carbonate resulted a reduction in the activity of acid and alkaline phosphatases in the cells of E. carteri. Experimental concentration of 8mg/l of washing soda for 192h yielded an increase in the physiological level of cellular apoptosis among the semigranulocytes and granulocytes of E. carteri, which was suggestive to possible shift in apoptosis mediated immunoprotection. The results were indicative of an undesirable shift in the immune status of sponge. Contamination of the freshwater aquifers by washing soda thus poses an alarming ecotoxicological threat to sponges.

Simultaneous inhibition of multiple oncogenic miRNAs by a multi-potent microRNA sponge.

The roles of oncogenic miRNAs are widely recognized in many cancers. Inhibition of single miRNA using antagomiR can efficiently knock-down a specific miRNA. However, the effect is transient and often results in subtle phenotype, as there are other miRNAs contribute to tumorigenesis. Here we report a multi-potent miRNA sponge inhibiting multiple miRNAs simultaneously. As a model system, we targeted miR-21, miR-155 and miR-221/222, known as oncogenic miRNAs in multiple tumors including breast and pancreatic cancers. To achieve efficient knockdown, we generated perfect and bulged-matched miRNA binding sites (MBS) and introduced multiple copies of MBS, ranging from one to five, in the multi-potent miRNA sponge. Luciferase reporter assay showed the multi-potent miRNA sponge efficiently inhibited 4 miRNAs in breast and pancreatic cancer cells. Furthermore, a stable and inducible version of the multi-potent miRNA sponge cell line showed the miRNA sponge efficiently reduces the level of 4 target miRNAs and increase target protein level of these oncogenic miRNAs. Finally, we showed the miRNA sponge sensitize cells to cancer drug and attenuate cell migratory activity. Altogether, our study demonstrates the multi-potent miRNA sponge is a useful tool to examine the functional impact of simultaneous inhibition of multiple miRNAs and proposes a therapeutic potential.

Donor mesenchymal stem cell-derived neural-like cells transdifferentiate into myelin-forming cells and promote axon regeneration in rat spinal cord transection.

IntroductionSevere spinal cord injury often causes temporary or permanent damages in strength, sensation, or autonomic functions below the site of the injury. So far, there is still no effective treatment for spinal cord injury. Mesenchymal stem cells (MSCs) have been used to repair injured spinal cord as an effective strategy. However, the low neural differentiation frequency of MSCs has limited its application. The present study attempted to explore whether the grafted MSC-derived neural-like cells in a gelatin sponge (GS) scaffold could maintain neural features or transdifferentiate into myelin-forming cells in the transected spinal cord.MethodsWe constructed an engineered tissue by co-seeding of MSCs with genetically enhanced expression of neurotrophin-3 (NT-3) and its high-affinity receptor tropomyosin receptor kinase C (TrkC) separately into a three-dimensional GS scaffold to promote the MSCs differentiating into neural-like cells and transplanted it into the gap of a completely transected rat spinal cord. The rats received extensive post-operation care, including cyclosporin A administrated once daily for 2 months.ResultsMSCs modified genetically could differentiate into neural-like cells in the MN + MT (NT-3-MSCs + TrKC-MSCs) group 14 days after culture in the GS scaffold. However, after the MSC-derived neural-like cells were transplanted into the injury site of spinal cord, some of them appeared to lose the neural phenotypes and instead transdifferentiated into myelin-forming cells at 8 weeks. In the latter, the MSC-derived myelin-forming cells established myelin sheaths associated with the host regenerating axons. And the injured host neurons were rescued, and axon regeneration was induced by grafted MSCs modified genetically. In addition, the cortical motor evoked potential and hindlimb locomotion were significantly ameliorated in the rat spinal cord transected in the MN + MT group compared with the GS and MSC groups.ConclusionGrafted MSC-derived neural-like cells in the GS scaffold can transdifferentiate into myelin-forming cells in the completely transected rat spinal cord.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0100-7) contains supplementary material, which is available to authorized users.

Integration of donor mesenchymal stem cell-derived neuron-like cells into host neural network after rat spinal cord transection

Functional deficits following spinal cord injury (SCI) primarily attribute to loss of neural connectivity. We therefore tested if novel tissue engineering approaches could enable neural network repair that facilitates functional recovery after spinal cord transection (SCT). Rat bone marrow-derived mesenchymal stem cells (MSCs), genetically engineered to overexpress TrkC, receptor of neurotrophin-3 (NT-3), were pre-differentiated into cells carrying neuronal features via co-culture with NT-3 overproducing Schwann cells in 3-dimensional gelatin sponge (GS) scaffold for 14 days in vitro. Intra-GS formation of MSC assemblies emulating neural network (MSC-GS) were verified morphologically via electron microscopy (EM) and functionally by whole-cell patch clamp recording of spontaneous post-synaptic currents. The differentiated MSCs still partially maintained prototypic property with the expression of some mesodermal cytokines. MSC-GS or GS was then grafted acutely into a 2 mm-wide transection gap in the T9-T10 spinal cord segments of adult rats. Eight weeks later, hindlimb function of the MSC-GS-treated SCT rats was significantly improved relative to controls receiving the GS or lesion only as indicated by BBB score. The MSC-GS transplantation also significantly recovered cortical motor evoked potential (CMEP). Histologically, MSC-derived neuron-like cells maintained their synapse-like structures in vivo; they additionally formed similar connections with host neurites (i.e., mostly serotonergic fibers plus a few corticospinal axons; validated by double-labeled immuno-EM). Moreover, motor cortex electrical stimulation triggered c-fos expression in the grafted and lumbar spinal cord cells of the treated rats only. Our data suggest that MSC-derived neuron-like cells resulting from NT-3-TrkC-induced differentiation can partially integrate into transected spinal cord and this strategy should be further investigated for reconstructing disrupted neural circuits.

Cell kinetics during regeneration in the sponge Halisarca caerulea: how local is the response to tissue damage?

Sponges have a remarkable capacity to rapidly regenerate in response to wound infliction. In addition, sponges rapidly renew their filter systems (choanocytes) to maintain a healthy population of cells. This study describes the cell kinetics of choanocytes in the encrusting reef sponge Halisarca caerulea during early regeneration (0–8 h) following experimental wound infliction. Subsequently, we investigated the spatial relationship between regeneration and cell proliferation over a six-day period directly adjacent to the wound, 1 cm, and 3 cm from the wound. Cell proliferation was determined by the incorporation of 5-bromo-2′-deoxyuridine (BrdU). We demonstrate that during early regeneration, the growth fraction of the choanocytes (i.e., the percentage of proliferative cells) adjacent to the wound is reduced (7.0 ± 2.5%) compared to steady-state, undamaged tissue (46.6 ± 2.6%), while the length of the cell cycle remained short (5.6 ± 3.4 h). The percentage of proliferative choanocytes increased over time in all areas and after six days of regeneration choanocyte proliferation rates were comparable to steady-state tissue. Tissue areas farther from the wound had higher rates of choanocyte proliferation than areas closer to the wound, indicating that more resources are demanded from tissue in the immediate vicinity of the wound. There was no difference in the number of proliferative mesohyl cells in regenerative sponges compared to steady-state sponges. Our data suggest that the production of collagen-rich wound tissue is a key process in tissue regeneration for H. caerulea, and helps to rapidly occupy the bare substratum exposed by the wound. Regeneration and choanocyte renewal are competing and negatively correlated life-history traits, both essential to the survival of sponges. The efficient allocation of limited resources to these life-history traits has enabled the ecological success and diversification of sponges.

Culturable associated-bacteria of the sponge Theonella swinhoei show tolerance to high arsenic concentrations.

Sponges are potent filter feeders and as such are exposed to high fluxes of toxic trace elements, which can accumulate in their body over time. Such is the case of the Red Sea sponge Theonella swinhoei, which has been shown to accumulate up to 8500 mg/Kg of the highly toxicelement arsenic. T. swinhoei is known to harbor a multitude of sponge-associated bacteria, so it is hypothesized that the associated-bacteria will be tolerant to high arsenic concentration. This study also investigates the fate of the arsenic accumulated in the sponge to test if the associated-bacteria have an important role in the arsenic accumulation process of their host, since bacteria are key players in the natural arsenic cycle. Separation of the sponge to sponge cells and bacteria enriched fractions showed that arsenic is accumulated by the bacteria. Sponge-associated, arsenic-tolerant bacteria were cultured in the presence of 5 mM of either arsenate or arsenite (equivalent to 6150 mg/Kg arsenic, dry weight). The 54 isolated bacteria were grouped to 15 operational taxonomic units (OTUs) and isolates belonging to 12 OTUs were assessed for tolerance to arsenate at increased concentrations up to 100 mM. Eight of the 12 OTUs tolerated an order of magnitude increase in the concentration of arsenate, and some exhibited external biomineralization of arsenic–magnesium salts. The biomineralization of this unique mineral was directly observed in bacteria for the first time. These results may provide an explanation for the ability of the sponge to accumulate considerable amounts of arsenic. Furthermore arsenic-mineralizing bacteria can potentially be used for the study of bioremediation, as arsenic toxicity affects millions of people worldwide.