No abstract

Pathological alterations of tau protein play a significant role in the emergence and progression of neurodegenerative disorders. Tauopathies are characterized by detachment of the tau protein from neuronal microtubules, and its subsequent aberrant hyperphosphorylation, aggregation and cellular distribution. The exact nature of tau protein species causing neuronal malfunction and degeneration is still unknown. In the present study, we used mice transgenic for human tau with the frontotemporal dementia with parkinsonism-associated P301S mutation. These mice are prone to develop fibrillar tau inclusions, especially in the spinal cord and brainstem. At the same time, cortical neurons are not as strongly affected by fibrillar tau forms, but rather by soluble tau forms. We took advantage of the possibility to induce formation of neurofibrillary tangles in a subset of these cortical neurons by local injection of preformed synthetic tau fibrils. By using chronic in vivo two-photon calcium imaging in awake mice, we were able for the first time to follow the activity of individual tangle-bearing neurons and compare it to the activity of tangle-free neurons over the disease course. Our results revealed strong reduction of calcium transient frequency in layer 2/3 cortical neurons of P301S mice, independent of neurofibrillary tangle presence. These results clearly point to the impairing role of soluble, mutated tau protein species present in the majority of the neurons investigated in this study.

Stable isotopes are used in ecology to track and disentangle different processes and pathways. Especially for studies focused on the gas exchange of plants, sensing techniques that offer oxygen (O2) and carbon dioxide (CO2) sensitivity with isotopic discrimination are highly sought after. Addressing this challenge, fiber-enhanced Raman gas spectroscopy is introduced as a fast optical technique directly combining 13CO2 and 12CO2 as well as 18O2 and 16O2 measurements in one instrument. We demonstrate how a new type of optical hollow-core fiber, the so-called revolver fiber, is utilized for enhanced Raman gas sensing. Carbon dioxide and oxygen isotopologues were measured at concentrations expected when using 13C- and 18O-labeled gases in plant experiments. Limits of detection have been determined to be 25 ppm for CO2 and 150 ppm for O2. The combination of measurements with different integration times allows the creation of highly resolved broadband spectra. With the help of calculations based on density functional theory, the line at 1512 cm–1 occurring in the oxygen spectrum is assigned to 18O16O. The relative abundances of the isotopologues 18O16O and nitrogen 15N14N were in good agreement with typical values. For CO2, fiber-enhanced Raman spectra show the Fermi diad and hotbands of 12C16O2, 13C16O2, and 12C18O16O. Several weak lines were observed, and the line at 1426 cm–1 was identified as originating from the (0 4 0 2) → (0 2 0 2) transition of 12C16O2. With the demonstrated sensitivity and discriminatory power, fiber-enhanced Raman spectroscopy is a possible alternative means to investigate plant metabolism, directly combining 13CO2 and 12CO2 measurements with 18O2 and 16O2 measurements in one instrument. The presented method thus has large potential for basic analytical investigations as well as for applications in the environmental sciences.