Coral Larvae Every Day: Leptastrea purpurea, a Brooding Species That Could Accelerate Coral Research

Sexually produced larvae are used in various fields of coral research. Because the vast majority of scleractinians reproduces only on one or few occasions per year through simultaneous release of gametes, and because an ex situ spawning induction is still very hard to achieve, great efforts are required to obtain planula larvae. Brooding corals have been used to harvest planulae although their larvae often differ in various traits from most spawning corals, e.g., in settlement behavior. Other cnidarians, such as Aiptasia spp., have been substituting for scleractinians in many aspects of coral research. However, organisms such as Aiptasia differ strongly from scleractinians, thus limiting the transferability of obtained results. This study examines the potential of Leptastrea purpurea as a reliable source of larvae for coral research. Larval output as well as settlement behavior of planulae was investigated. Our results show that colonies of L. purpurea released a daily average of 3.7 (±0.2) larvae during a period of 65 days, thus allowing continual access to planula larvae. We collected a total of 58127 larvae from our broodstock of 243 colonies. Larval settlement is induced by the same and/or similar cues as in many spawning species which increases the transferability of conclusions. We discuss the utility of L. purpurea for research on scleractinian physiology, ecology and larval settlement and conclude that L. purpurea is a well-suited organism to accelerate progress in many fields of coral research

Development of 3D human intestinal equivalents for substance testing in microliter-scale on a multi-organ-chip : From 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better Health Lille, France. 23-26 June 2013

First published by BioMed Central: Jaenicke, Annika; Tordy, Dominique; Groeber, Florian; Hansmann, Jan; Nietzer, Sarah; Tripp, Carolin; Walles, Heike; Lauster, Roland; Marx, Uwe: Development of 3D human intestinal equivalents for substance testing in microliter-scale on a multi-organ-chip. - In: BMC Proceedings. - ISSN 1753-6561 (online). - 7 (2013), suppl. 6, P65. - doi:10.1186/1753-6561-7-S6-P65

Connecting Cancer Pathways to Tumor Engines: A Stratification Tool for Colorectal Cancer Combining Human In Vitro Tissue Models with Boolean In Silico Models

To improve and focus preclinical testing, we combine tumor models based on a decellularized tissue matrix with bioinformatics to stratify tumors according to stage-specific mutations that are linked to central cancer pathways. We generated tissue models with BRAF-mutant colorectal cancer (CRC) cells (HROC24 and HROC87) and compared treatment responses to two-dimensional (2D) cultures and xenografts. As the BRAF inhibitor vemurafenib is—in contrast to melanoma—not effective in CRC, we combined it with the EGFR inhibitor gefitinib. In general, our 3D models showed higher chemoresistance and in contrast to 2D a more active HGFR after gefitinib and combination-therapy. In xenograft models murine HGF could not activate the human HGFR, stressing the importance of the human microenvironment. In order to stratify patient groups for targeted treatment options in CRC, an in silico topology with different stages including mutations and changes in common signaling pathways was developed. We applied the established topology for in silico simulations to predict new therapeutic options for BRAF-mutated CRC patients in advanced stages. Our in silico tool connects genome information with a deeper understanding of tumor engines in clinically relevant signaling networks which goes beyond the consideration of single drivers to improve CRC patient stratification

Gen-Umwelt-Interaktionen in Serotonin-Transporter-Knockout-Mäusen - Wie Stress die Genexpression und die neuronale Morphologie beeinflusst

Serotonin (5-HT) is an important modulator of many physiological, behavioural and developmental processes and it plays an important role in stress coping reactions. Anxiety disorders and depression are stress-related disorders and they are associated with a malfunction of the 5-HT system, in which the 5-HT transporter (5-HTT) plays an important role. 5-Htt knockout (KO) mice represent an artificially hyperserotonergic environment, show an increased anxiety-like behaviour and seem to be a good model to investigate the role of the 5-HT system concerning stress reactions and anxiety disorders. As synaptic proteins (SPs) seem to be involved in stress reactions, the effect of acute immobilization stress on the expression of the three SPs Synaptotagmin (Syt) I, Syt IV and Syntaxin (Stx) 1A was studied in the 5-Htt KO mouse model as well as the expression of the two immediate early genes (IEGs) FBJ osteosarcoma oncogene (c-Fos) and fos-like antigen 2 (Fra-2). Additionally, the expression of the corticotrophin releasing hormone (CRH) and its two receptors CRHR1 and CRHR2 was investigated as part of the hypothalamic-pituitary-adrenal (HPA) stress system. Based on gender- and genotype-dependent differences in corticosterone levels, expression differences in the brain were investigated by performing a quantitative real time-PCR study using primer pairs specific for these SPs and for the IEGs c-Fos and Fra-2 in five different brain regions in 5-Htt KO and 5-Htt wild-type (WT) mice. Mainly gender-dependent differences could be found and weaker stress effects on the expression of SPs could be demonstrated. Regarding the expression of IEGs, stress-, gender- and genotype-dependent differences were found mainly in the hypothalamus. Also in the hypothalamus, gender effects were found concerning the expression of CRH and its both receptors. Additionally, in a second study, male 5-Htt WT and male 5-Htt deficient mice were subjected to a resident-intruder-paradigm which stresses the animals through a loser experience. The morphological changes of neurons were subsequently analyzed in Golgi-Cox-stained sections of limbic brain areas in stressed and unstressed animals of both genotypes using the computer-based microscopy system Neurolucida (Microbrightfield, Inc.). While no differences concerning dendritic length, branching patterns and spine density were found in the hippocampus and no differences concerning dendritic length and branching patterns could be shown in the cingulate cortex (CG), pyramidal neurons in the infralimbic cortex (IL) of stressed 5-Htt WT mice displayed longer dendrites compared to unstressed 5-Htt WT mice. The results indicate that, although in this model drastic alterations of neuronal morphology are absent, subtle changes can be found in specific brain areas involved in stress- and anxiety-related behaviour which may represent neural substrates underlying behavioural phenomena.Serotonin (5-HT) ist ein wichtiger Modulator vieler physiologischer, verhaltensbiologischer und entwicklungsbiologischer Vorgänge und spielt zudem eine wichtige Rolle bei der Stressbewältigung. Angsterkrankungen und Depression sind stressbedingte Störungen und sie sind mit einer Dysfunktion des serotonergen Systems assoziiert, in dem der Serotonintransporter (5-HTT) eine wichtige Funktion einnimmt. 5-Htt Knockout (KO)-Mäuse haben reduzierte Serotoninkonzentrationen im Gehirn, zeigen erhöhtes Angst-ähnliches Verhalten und scheinen ein gutes Modell für die Erforschung der Rolle des serotonergen Systems in Bezug auf Stress-Reaktionen und Angsterkrankungen darzustellen. Da synaptische Proteine (SPs) in die Stress-Reaktion involviert zu sein scheinen, wurden die Auswirkungen von akutem Immobilizierungsstress auf die Expression der drei SPs Synaptotagmin (Syt) I, Syt IV und Syntaxin (Stx) 1A in diesem 5-Htt KO-Maus-Modell untersucht. Ebenso wurde die Expression der zwei „immediate early genes“ (IEGs) FBJ osteosarcoma oncogene (c-Fos) and fos-like antigen 2 (Fra-2) unter die Lupe genommen. Außerdem wurde die Expression des „Corticotrophin Releasing Hormone“ (CRH) sowie seiner beiden Rezeptoren CRHR1 and CRHR2, als Teil des Hypothalamus- Hypophysen-Nebennieren-(HPA)-Systems, analysiert. Basierend auf Geschlechts- und Genotyp-spezifischen Unterschieden der Kortikosteron-Konzentrationen im Blut der Tiere wurden die Expressionslevel dieser SPs und der beiden IEGs mittels quantitativer Real-Time (qRT)-PCR in fünf verschiedenen Gehirnregionen von 5-Htt KO- und 5-Htt-Mäusen mit wildtypischer (WT) Gen-Konstitution untersucht. Dabei konnten vor allem Geschlechts-spezifische Unterschiede in der Genexpression gezeigt werden und es konnte ein im Vergleich dazu schwächerer Einfluss des akuten Immobilisierungs-Stresses auf die Genexpression nachgewiesen werden. Die Expression der IEGs wurde durch Stress, Geschlecht und Genotyp vor allem im Hypothalamus beeinflusst. Ebenfalls im Hypothalamus konnte der Einfluss des Geschlechts auf die Expression des CRH und seiner beiden Rezeptoren gezeigt werden. In einer zweiten Studie wurden männliche 5-Htt KO-Mäuse sowie 5-Htt WT-Mäuse dem „Resident-Intruder-Paradigma“ unterzogen, in welchem die Tiere mittels einer mehrfachen Verlierer-Erfahrung gestresst wurden. Morphologische Veränderungen von Neuronen limbischer Gehirnareale wurden daraufhin an Golgi-Cox-gefärbten Gehirnschnitten dieser gestressten und ungestressten 5-Htt KO- und 5-Htt WT-Tiere mittels des Computer-gestützten Mikroskop-Systems Neurolucida (Microbrightflield, Inc.) analysiert. Während keine Unterschiede bezüglich der Länge des dendritischen Materials, des Verzweigungsmusters und der Spinedichte im Hippocampus gefunden werden konnten und keine Unterschiede in Länge des dendritischen Materials und des Verzweigungsmusters in der Area cinguli (CG) gezeigt werden konnten, wiesen Pyramidenzellen im infralimbischen Kortex (IL) von gestressten 5-Htt WT-Mäusen längere Dendriten auf als die entsprechenden Zellen in den ungestressten Tieren desselben Genotyps. Diese Ergebnisse zeigen, dass, obwohl in diesen Tieren keine drastischen stressbedingten Änderungen der neuronalen Morphologie vorliegen, doch subtile Änderungen der neuronalen Morphologie stress- und angstinvolvierter Gehirnareale gefunden werden können. Diese Änderungen können die neuronale Basis verschiedenster Verhaltens-Phänomene darstellen

Mono- and multispecies biofilms from a crustose coralline alga induce settlement in the scleractinian coral Leptastrea purpurea

Microorganisms have been reported to induce settlement in various marine invertebrate larvae but their specificity of inductive capacities for the settlement of coral larvae remains poorly understood. In this study, we isolated 56 microbial strains from the crustose coralline alga (CCA) Hydrolithon reinboldii using five different media either with or without additional antibiotics and/or spiked CCA extract. We tested the isolates for their potential to induce settlement behavior in larvae of the brooding scleractinian coral Leptastrea purpurea. From these 56 CCA-associated microbial strains, we identified six bacterial classes and 18 families. The culturable bacterial community associated with H. reinboldii was dominated by Gammaproteobacteria, Actinobacteria, and Alphaproteobacteria while the Illumina MiSeq analysis showed that the culture-independent bacterial community was dominated by Gammaproteobacteria, Alphaproteobacteria, and Flavobacteria. Furthermore, we found no correlation between inductive settlement capacities and phylogenetic relationships. Instead, settlement behavior of L. purpurea larvae was induced by specific isolated species. Strains #1792 (Pseudovibrio denitrificans), #1678 (Acinetobacter pittii), #1633 (Pseudoalteromonas phenolica), #1772 (Marine bacterium LMG1), #1721 (Microbulbifer variabilis), and #1783 (Pseudoalteromonas rubra) induced settlement behavior in coral larvae at mostly high and significant levels (>= 40%) but the remaining isolates strongly varied in their activity. Multispecies biofilms consisting of four strains (#1792, #1678, #1633, and #1721) were observed to synergistically increase settlement behavior levels (> 90%); however, the addition of #1772 to the multispecies biofilms negatively affected coral larvae and resulted in a total loss of inducing activity. The findings provide new insights into the role of bacteria in the settlement process of scleractinian corals and may help to identify the true nature of bacteria-derived morphogenic cues