ECEL1 mutation implicates impaired axonal arborization of motor nerves in the pathogenesis of distal arthrogryposis.

The membrane-bound metalloprotease endothelin-converting enzyme-like 1 (ECEL1) has been newly identified as a causal gene of a specific type of distal arthrogryposis (DA). In contrast to most causal genes of DA, ECEL1 is predominantly expressed in neuronal cells, suggesting a unique neurogenic pathogenesis in a subset of DA patients with ECEL1 mutation. The present study analyzed developmental motor innervation and neuromuscular junction formation in limbs of the rodent homologue damage-induced neuronal endopeptidase (DINE)-deficient mouse. Whole-mount immunostaining was performed in DINE-deficient limbs expressing motoneuron-specific GFP to visualize motor innervation throughout the limb. Although DINE-deficient motor nerves displayed normal trajectory patterns from the spinal cord to skeletal muscles, they indicated impaired axonal arborization in skeletal muscles in the forelimbs and hindlimbs. Systematic examination of motor innervation in over 10 different hindlimb muscles provided evidence that DINE gene disruption leads to insufficient arborization of motor nerves after arriving at the skeletal muscle. Interestingly, the axonal arborization defect in foot muscles appeared more severe than in other hindlimb muscles, which was partially consistent with the proximal-distal phenotypic discordance observed in DA patients. Additionally, the number of innervated neuromuscular junction was significantly reduced in the severely affected DINE-deficient muscle. Furthermore, we generated a DINE knock-in (KI) mouse model with a pathogenic mutation, which was recently identified in DA patients. Axonal arborization defects were clearly detected in motor nerves of the DINE KI limb, which was identical to the DINE-deficient limb. Given that the encoded sequences, as well as ECEL1 and DINE expression profiles, are highly conserved between mouse and human, abnormal arborization of motor axons and subsequent failure of NMJ formation could be a primary cause of DA with ECEL1 mutation.

Generation of App knock-in mice reveals deletion mutations protective against Alzheimer's disease-like pathology.

Although, a number of pathogenic mutations have been found for Alzheimer's disease (AD), only one protective mutation has been identified so far in humans. Here we identify possible protective deletion mutations in the 3'-UTR of the amyloid precursor protein (App) gene in mice. We use an App knock-in mouse model carrying a humanized Aß sequence and three AD mutations in the endogenous App gene. Genome editing of the model zygotes using multiple combinations of CRISPR/Cas9 tools produces genetically mosaic animals with various App 3'-UTR deletions. Depending on the editing efficiency, the 3'-UTR disruption mitigates the Aß pathology development through transcriptional and translational regulation of APP expression. Notably, an App knock-in mouse with a 34-bp deletion in a 52-bp regulatory element adjacent to the stop codon shows a substantial reduction in Aß pathology. Further functional characterization of the identified element should provide deeper understanding of the pathogenic mechanisms of AD.

Mouse Ability to Perceive Subjective Contours.

In contrast to the previously held notion that mice have a weak visual system, it is now generally accepted that mice can perceive rather complicated figures in various contexts such as in cognitive experiments and in social settings. Here, we show that mice could even be capable of perceiving a visual illusion--subjective contours. This illusion requires the visual system to compensate for a lack of visual information in compressed 2D images on the retina. In this experiment, we trained mice to respond appropriately to a rectangle-shaped rewarded figure of specific orientation in a two-choice visual discrimination task with a touchscreen monitor. In Transfer Test 1, mice could discriminate illusory rectangle-shaped figures significantly as compared with a figure, which did not induce illusory figures. In Transfer Test 2, the choice rate of targets decreased with imperfect illusory figures, which produced weak perception of rotated or deficient inducers. Moreover, in Transfer Test 3, mice could not discriminate the low-resolution illusory figure, which also induced weak perception. These results demonstrated the possibility that mice might be useful for investigating fundamental properties of the neural visual system.