Biology subjects

Thiele, T.

Publications and source records attributed to Thiele, T..

3 recordsLinked to original sources

bioRxiv2024

Haemolymphatic tissues of captive boa constrictor (Boa constrictor): morphological features in healthy individuals and with Boid Inclusion Body Disease

Knowledge on the structure and composition of the haematopoietic tissue (HT) is essential to understand the basic immune functions of the immune system in any species. For reptiles, it is extremely limited, hence we undertook an in-depth in situ investigation of the HT (bone marrow, thymus, spleen, lymphatic tissue of the alimentary tract) in the common boa (Boa constrictor). We also assessed age- and disease-related changes, with a special focus on Boid Inclusion Body Disease, a highly relevant reptarenavirus-associated disease in boid snakes. The HT was subjected to gross, histological and ultrastructural examination, including special stains, immunohistochemistry, in situ hybridization and morphometric analyses. In general, the HT was dominated by T cells and lacked a clear structural organization, such as follicle formation. BIBD was associated with significantly higher cellularity and a granulomatous response in the spleen, and the presence of virus-infected haematopoietic cells in the bone marrow, suggesting the latter as a persistent source of viremia. HighlightsO_LILymphatic tissues of B. constrictor lack organization and are dominated by T cells C_LIO_LIThe thymus of B. constrictor shows slow age-related involution C_LIO_LIBIBD-positive boas show a granulomatous response in the spleen C_LIO_LIReptarenaviruses infect haematopoietic cells in the bone marrow C_LIO_LIReptarenavirus infected bone marrow cells might represent a source of viremia C_LI

pathology↗

bioRxiv2024

Dynamic regulation of CeA gene expression during acute and protracted abstinence from chronic binge drinking of male and female C57BL/6J mice.

Binge alcohol consumption is a major risk factor for developing Alcohol Use Disorder (AUD) and is associated with alcohol-related problems like accidental injury, acute alcohol poisoning, and black-outs. While there are numerous brain regions that have been shown to play a role in this AUD in humans and animal models, the central nucleus of the amygdala (CeA) has emerged as a critically important locus mediating binge alcohol consumption. In this study, we sought to understand how relative gene expression of key signaling molecules in the CeA changes during different periods of abstinence following bouts of binge drinking. To test this, we performed drinking in the dark (DID) on two separate cohorts of C57BL/6J mice and collected CeA brain tissue at one day (acute) and 7 days (protracted) abstinence after DID. We used qRTPCR to evaluate relative gene expression changes of 25 distinct genes of interest related to G protein-coupled receptors (GPCRs), neuropeptides, ion channel subunits, and enzymes that have been previously implicated in AUD. Our findings show that during acute abstinence CeA punches collected from female mice had upregulated relative mRNA expression of the gamma-aminobutyric acid receptor subunit alpha 2 (Gabra2), and the peptidase, angiotensinase c (Prcp). CeA punches from male mice at the same time point in abstinence had upregulated relative mRNA encoding for neuropeptide-related molecules, neuropeptide Y (Npy) and somatostatin (Sst), as well as the neuropeptide Y receptor Y2 (Npyr2) but downregulated, Glutamate ionotropic receptor NMDA type subunit 1 (Grin1). After protracted abstinence CeA punches collected from female mice had increased mRNA expression of corticotropin releasing hormone (Crh) and Npy. While CeA punches collected from male mice at the same timepoint had upregulated relative mRNA expression of Npy2r and downregulated mRNA expression of Gabra2, Grin1 and opioid receptor kappa 1 (Oprk1). Our findings support that there are differences in how the CeA of male and female respond to binge-alcohol exposure, highlighting the need to understand the implications of such differences in the context of AUD and binge drinking behavior.

neuroscience↗

bioRxiv2023

Immobilization of Lipophilic and Amphiphilic Biomarker on Hydrophobic Microbeads

BackgroundLipids and amphiphilic molecules are ubiquitous and play a central role in cell signalling, cell membrane structure, and lipid transport in the human body. However, they also appear in many diseases such as atherosclerosis, cardiovascular diseases, infections, inflammatory diseases, cancer, and autoimmune diseases. Thus, it is necessary to have detection systems for lipids and amphiphilic molecules. Microbeads can be one of these systems for the simultaneous detection of different lipophilic biomarkers. MethodsBased on the fundamentals of microbead development, novel hydrophobic microbeads were produced. These not only have a hydrophobic surface, but are also fluorescently encoded and organic solvent resistant. The challenge after the development of the hydrophobic microbeads was to immobilise the amphiphilic molecules, in this study phospholipids, on the microbead surface in an oriented direction. After successful immobilisation of the biomarkers, a suitable antibody based detection assay was established. ResultsBy passive adsorption, the phospholipids cardiolipin, phosphatidylethanolamine and phosphatidylcholine could be bound to the microbead surface. With the application of the enzymes phospholipase A2 and phospholipase C, the directional binding of the phospholipids to the microbead surface was demonstrated. The detection of directional binding indicated the need for the hydrophobic surface. Microbeads with no hydrophobic surface bound the phospholipids non-directionally (with the hydrophilic head) and were thus no longer reactively accessible for detection. ConclusionWith the newly developed hydrophobic, dual coded and solvent stable microbeads it is possible to bind amphiphilic biomolecules directionally onto the microbead surfaces.

biochemistry↗