New developments in celiac disease treatments.

Celiac disease (CeD), an autoimmune disorder triggered by gluten, affects around 1% of the global population. Standard treatment is a strict gluten-free diet (GFD), which poses significant challenges due to dietary restrictions, cross-contamination and subsequent persistent intestinal inflammation. This underscores the need for new treatment options addressing the complex pathophysiology of CeD.

Cortical circuit dysfunction in a mouse model of alpha-synucleinopathy .

Considerable fluctuations in cognitive performance and eventual dementia are an important characteristic of alpha-synucleinopathies, such as Parkinson's disease and Lewy Body dementia and are linked to cortical dysfunction. The presence of misfolded and aggregated alpha-synuclein in the cerebral cortex of patients has been suggested to play a crucial role in this process. However, the consequences of a-synuclein accumulation on the function of cortical networks at cellular resolution are largely unknown.

In vivo imaging reveals reduced activity of neuronal circuits in a mouse tauopathy model.

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 vivo Ca imaging of astrocytic microdomains reveals a critical role of the amyloid precursor protein for mitochondria.

The investigation of amyloid precursor protein (APP) has been mainly confined to its neuronal functions, whereas very little is known about its physiological role in astrocytes. Astrocytes exhibit a particular morphology with slender extensions protruding from somata and primary branches. Along these fine extensions, spontaneous calcium transients occur in spatially restricted microdomains.

Beta-Site Amyloid Precursor Protein Cleaving Enzyme 1 Inhibition Impairs Synaptic Plasticity via Seizure Protein 6.

Background: Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) is a promising drug target for the treatment of Alzheimer's disease. Prolonged BACE1 inhibition interferes with structural and functional synaptic plasticity in mice, most likely by altering the metabolism of BACE1 substrates. Seizure protein 6 (SEZ6) is predominantly cleaved by BACE1, and Sez6 knockout mice share some phenotypes with BACE1 inhibitor-treated mice.

Generation and Screening of Transgenic Mice with Neuronal Labeling Controlled by Thy1 Regulatory Elements.

Major progress has been made using in vivo imaging in mice to study mammalian nervous system development, plasticity, and disease. This progress has depended in part on the wide availability of two-photon microscopy, which is capable of penetrating deep into scattering tissue. Equally important, however, is the generation of suitable transgenic mouse models, which provide a "Golgi staining"-like labeling of neurons that is sparse and bright enough for in vivo imaging.

Generation of Thy1 Constructs for Pronuclear Injection.

With easy access to core facilities or commercial providers of pronuclear injections, generating simple Thy1-XFP transgenic mice (where XFP stands for any fluorescent protein) is now a possibility even for small laboratories. The generation of new Thy1 transgenic lines generally consists of five steps: (1) engineering and characterization of the desired fluorescent reporter protein, (2) cloning of the reporter protein into the Thy1 vector, (3) linearization and purification of the new Thy1 construct, (4) pronuclear injection to generate founders, and (5) screening of founder progeny to establish transgenic lines. Here, we provide a protocol for Steps 2 and 3.

Imaging Acute Neuromuscular Explants from Thy1 Mouse Lines.

Because core facilities that generate transgenic founder mice for a reasonable fee are now available at most major research institutions, generating new Thy1-XFP transgenic animals (in which XFP stands for any fluorescent protein) is an option even for relatively small laboratories. Here, we provide a protocol for screening offspring of Thy1 transgenic founders. Acute neuromuscular explants are obtained from 3-wk-old F1 mice that have been produced by crossing Thy1 transgenic founders and commercially obtained inbred mice.

Generation of Tissue Sections for Screening Thy1 Mouse Lines.

New generations of Thy1-XFP transgenic mice (where XFP stands for any fluorescent protein) can now be readily generated, given the availability of core facilities or commercial providers of Thy1 pronuclear injections. Here, we provide a protocol for screening founder progeny. Transcardial perfusion is performed on 3-wk-old F1 mice that have been produced by crossing Thy1 transgenic founders and commercially obtained inbred mice.

Pervasive axonal transport deficits in multiple sclerosis models.

Impaired axonal transport can contribute to axon degeneration and has been described in many neurodegenerative diseases. Multiple sclerosis (MS) is a common neuroinflammatory disease, which is characterized by progressive axon degeneration-whether, when, and how axonal transport is affected in this condition is unknown. Here we used in vivo two-photon imaging to directly assay transport of organelles and the stability of microtubule tracks in individual spinal axons in mouse models of MS.

An assay to image neuronal microtubule dynamics in mice.

Microtubule dynamics in neurons play critical roles in physiology, injury and disease and determine microtubule orientation, the cell biological correlate of neurite polarization. Several microtubule binding proteins, including end-binding protein 3 (EB3), specifically bind to the growing plus tip of microtubules. In the past, fluorescently tagged end-binding proteins have revealed microtubule dynamics in vitro and in non-mammalian model organisms.

Multiparametric optical analysis of mitochondrial redox signals during neuronal physiology and pathology in vivo.

Mitochondrial redox signals have a central role in neuronal physiology and disease. Here we describe a new optical approach to measure fast redox signals with single-organelle resolution in living mice that express genetically encoded redox biosensors in their neuronal mitochondria. Moreover, we demonstrate how parallel measurements with several biosensors can integrate these redox signals into a comprehensive characterization of mitochondrial function.

Sequential photo-bleaching to delineate single Schwann cells at the neuromuscular junction.

Sequential photo-bleaching provides a non-invasive way to label individual SCs at the NMJ. The NMJ is the largest synapse of the mammalian nervous system and has served as guiding model to study synaptic structure and function. In mouse NMJs motor axon terminals form pretzel-like contact sites with muscle fibers.

Axonal transport deficits and degeneration can evolve independently in mouse models of amyotrophic lateral sclerosis.

Axonal transport deficits have been reported in many neurodegenerative conditions and are widely assumed to be an immediate causative step of axon and synapse loss. By imaging changes in axonal morphology and organelle transport over time in several animal models of amyotrophic lateral sclerosis (ALS), we now find that deficits in axonal transport of organelles (mitochondria, endosomes) and axon degeneration can evolve independently. This conclusion rests on the following results: (i) Axons can survive despite long-lasting transport deficits: In the SOD(G93A) model of ALS, transport deficits are detected soon after birth, months before the onset of axon degeneration.

Changes of behavioral parameters during long-term food restriction in middle-aged Wistar rats.

Food restriction (FR) has a beneficial effect on aging process and exerts a significant effect on the responses of rodents to standard behavioral tasks. The aim of this study was to assess the cumulative influence of FR on the behavioral and biochemical parameters in Wistar rats. Six-month-old rats were subjected to restrictive feeding (50% of the daily food intake, every-other-day feeding regimen) for one month or for six months until ages of 7 and 12months, respectively.

Behavioral and biochemical effects of various food-restriction regimens in the rats.

In this paper we describe the effects of six different food restriction (FR) regimens on amphetamine (AMPH)-induced locomotor and nonlocomotor activities in male rats. Changes in serum corticosterone (CORT), insulin and glucose levels were also examined. Each regimen was implemented through different daily food allowance (50%, 25% and 12.