Biology subjects

Eriksen, J.

Publications and source records attributed to Eriksen, J..

6 recordsLinked to original sources

bioRxiv2023

Adaptor Protein-3 Produces Synaptic Vesicles that Release Phasic Dopamine

The burst firing of midbrain dopamine neurons releases a phasic dopamine signal that mediates reinforcement learning. At many synapses, however, high firing rates deplete synaptic vesicles (SVs), resulting in synaptic depression that limits release. What accounts for the increased release of dopamine by stimulation at high frequency? We find that adaptor protein-3 (AP-3) and its coat protein VPS41 promote axonal dopamine release by targeting vesicular monoamine transporter VMAT2 to the axon rather than dendrites. AP-3 and VPS41 also produce SVs that respond preferentially to high frequency stimulation, independent of their role in axonal polarity. In addition, conditional inactivation of VPS41 in dopamine neurons impairs reinforcement learning, and this involves a defect in the frequency dependence of release rather than the amount of dopamine released. Thus, AP-3 and VPS41 promote the axonal polarity of dopamine release but enable learning by producing a novel population of SVs tuned specifically to high firing frequency that confers the phasic release of dopamine. Significance statementReinforcement learning requires the phasic dopamine produced by burst firing but synaptic vesicle depletion limits the ability to convey information at high firing rates. We now find that AP-3 has two independent roles in dopamine release. First, AP-3 confers the axonal polarity of dopamine release by targeting vesicular monoamine transporter 2 (VMAT2) to the axon. Second, AP-3 acting locally at the nerve terminal produces synaptic vesicles that respond specifically to high frequency stimulation. Consistent with this, loss of AP-3 impairs reinforcement learning and this reflects the defect in release at high frequency, not the reduction in axonal dopamine.

neuroscience↗

bioRxiv2022

Allosteric Regulation of a Synaptic Vesicle Glutamate Transporter

Concentration of neurotransmitter inside synaptic vesicles (SVs) underlies the quantal nature of synaptic transmission. In contrast to many transporters, SV uptake of the principal excitatory neurotransmitter glutamate is driven by membrane potential. To prevent nonquantal efflux of glutamate after SV exocytosis, the vesicular glutamate transporters (VGLUTs) are allosterically inhibited by the neutral pH of the synaptic cleft. We have now determined high-resolution structures of VGLUT2 with a cyclic analog of glutamate bound that defines the mechanism of substrate recognition, a positively charged cytoplasmic vestibule that electrostatically attracts the negatively charged substrate, and modification by palmitoylation that promotes retrieval of the transporter after exocytosis. The structure also incorporates an extensive, cytoplasmic network of electrostatic interactions that acts as a gate. Functional analysis shows how this cytoplasmic gate confers the allosteric requirement for lumenal H+ required to restrict VGLUT activity to SVs.

biochemistry↗

bioRxiv2021

Allosteric Inhibition of a Vesicular Glutamate Transporter by an Isoform-Specific Antibody

The role of glutamate in excitatory neurotransmission depends on its transport into synaptic vesicles by the vesicular glutamate transporters (VGLUTs). The three VGLUT isoforms exhibit a complementary distribution in the nervous system and the knockout of each produces severe, pleiotropic neurological effects. However, the available pharmacology lacks sensitivity and specificity, limiting the analysis of both transport mechanism and physiological role. To develop new molecular probes for the VGLUTs, we raised six mouse monoclonal antibodies to VGLUT2. All six bind to a structured region of VGLUT2, five to the luminal face and one to the cytosolic. Two are specific to VGLUT2 whereas the other four bind to both VGLUT1 and 2; none detect VGLUT3. Antibody 8E11 recognizes an epitope spanning the three extracellular loops in the C-domain that explains the recognition of both VGLUT1 and 2 but not VGLUT3. 8E11 also inhibits both glutamate transport and the VGLUT-associated chloride conductance. Since the antibody binds outside the substrate recognition site, it acts allosterically to inhibit function presumably by restricting conformational changes. The isoform specificity also shows that allosteric inhibition provides a mechanism to distinguish between closely related transporters.

biochemistry↗

bioRxiv2021

Early Detection of Tau Pathology

While a definitive Alzheimers disease (AD) diagnosis remains a post-mortem exercise, the ATN Research Framework proposed by the National Institute on Aging and the Alzheimers Association utilizes a score representing the presence of amyloid deposits (A), tau deposits (T) and neuronal degeneration markers (N), with A+T+ necessary for a positive diagnosis. Current detection of tau pathology lags amyloid detection by years and by the time both markers are detected the disease is fairly advanced. We describe the development of a new generation of molecular imaging probes for in vivo detection of cells undergoing abnormal phosphorylation representing the initial stages of pTau pathology, potentially enabling a very early stage diagnosis of AD. We describe a novel nanoparticle formulation that binds such abnormally phosphorylating cells in a mouse model of tau pathology, enabling in vivo visualization of the hyperphosphorylative state by magnetic resonance imaging. Our results demonstrate the potential of this novel platform to identify a correlative marker signifying the development of future tau pathology, and has implications for early-stage diagnosis of Alzheimers disease.

neuroscience↗

bioRxiv2021

Prostacyclin as a Negative Regulator of Angiogenesis in the Neurovasculature

In this study, multiple measures of angiogenic processes were assessed in murine brain endothelial (bEnd.3) cells after exposure to the stable prostacyclin analog, iloprost. Additionally, changes in the {gamma}-secretase enzyme were evaluated after activation of prostacyclin signaling using PGI2 overexpressing mouse brain tissue and immunohistology studies in bEnd.3 cells. A three-dimensional assay of tube formation revealed that iloprost inhibits normal formation by significantly reduced tube lengths and vessel mesh area. The iloprost-mediated inhibition of tube-like structures was ameliorated by a specific IP-receptor antagonist, CAY10449. Reductions in wound healing were observed with iloprost application in a dose-dependent manner and this effect was reversed using CAY10449. Iloprost did not exhibit anti-proliferative effects in the bEnd.3 cells. When subjected to a Transwell assay to evaluate changes in trans-epithelial electrical resistance (TEER), bEnd.3 cells displayed reduced TEER values in the presence of iloprost an effect that lasted over prolonged periods (24 hours). Again, CAY10449 was able to reverse iloprost-mediated reductions in TEER value. Surprisingly, the adenylyl cyclase activator, forskolin, produced higher TEER values in the bEnd.3 cells over the same time. The TEER results suggest that iloprost may not activating the Gs protein of the IP receptor to increase cAMP levels given by the opposing results seen with iloprost and forskolin. In terms of {gamma}-secretase expression, PGI2 overexpression in mice increased the expression of the APH-1 subunit in the hippocampus and cortex. In bEnd.3 cells, iloprost application slightly increased APH-1 subunit expression measured by western blot and interrupted the colocalization of Presenilin 1 and APH-1 subunits using immunohistochemistry. The results suggest that prostacyclin signaling within bEnd.3 cells is anti-angiogenic and further downstream events have effects on the expression and most likely the activity of the A{beta} cleaving enzyme, {gamma}-secretase.

neuroscience↗

bioRxiv2019

The Swedish example of food animal production without extensive use of antibiotics - or \"healthy animals do not need antibiotics\".

ObjectiveTo describe how stakeholders at different levels in food animal production in Sweden work to contain antibiotic resistance, with a special focus on poultry production. The stakeholders perceptions of antibiotic resistance and awareness of the One Health concept were also studied.\n\nMethodsThis is an interview study with thirteen informants. They represent policymakers, trade organisations, and veterinarians and farmers in the poultry industry. Interview transcripts were analysed using content analysis. The analysis continued until a latent theme emerged, and then the content was rearranged in four domains.\n\nFindingsA latent theme \"Working in unison\" emerged, based on the consistency expressed by the informants when they discussed antibiotic resistance, use of antibiotics and food animal production methods. The theme was built on four domains, representing the content of the interviews: Knowledge and engagement; Cooperation; Animal health concept; and Development in balance with economic prerequisites. The work for healthy animals started in Sweden already in the 1920-ies and continued step by step in cooperation and with support from the government. In 1986 Sweden became the first country to ban antibiotics for growth promotion. Veterinarians were considered important drivers of processes by spreading knowledge and working close to the farmers. Farmers felt involved in the development of production methods. The One Health concept was well known among stakeholders working at national level but not among veterinarians in production or farmers.\n\nConclusionsSweden has come far in work to contain antibiotic resistance in the animal sector by practicing restrictive use of antibiotics in food animal production. This practise is based on a long tradition of cooperation among stakeholders, from policymakers to farmers, and with a primary focus on animal health and welfare.

microbiology↗