CRMP4 is required for the positioning and maturation of newly generated neurons in adult mouse hippocampus.

Adult neurogenesis is a phenomenon in which neural stem cells differentiate and mature to generate new neurons in the adult brain. In mammals, the sites where adult neurogenesis occurs are limited to the subgranular zone (SGZ) of the hippocampal dentate gyrus and the subventricular zone. In the hippocampus, newly generated neurons migrate into the granule cell layer (GCL) and are integrated into neural circuits. Previous studies have revealed that CRMP4, a member of the CRMP family, is expressed in immature neurons in the hippocampal SGZ of the adult brain. However, the role of CRMP4 in adult neurogenesis is unknown. To study the role of CRMP4 in hippocampal adult neurogenesis, we compared adult neurogenesis between wild type and CRMP4-/- mice. In CRMP4-/- mice, the number of doublecortin (DCX)-positive cells was comparable to that in wild-type mice, and some DCX-positive cells were ectopically located in the granule cell layer, suggesting that CRMP4 is involved in the migration of adult neurogenesis. In addition, the number of calretinin-positive new neurons in the SGZ was significantly increased, whereas the number of EdU/NeuN-double positive neurons was decreased in CRMP4-/- mice, suggesting that CRMP4 plays an important role in neuronal maturation. Because CRMP4 is expressed in immature neurons, its expression may regulate the migration from the SGZ to the GCL during neuronal maturation in hippocampal adult neurogenesis.

Regulation of axon pruning of mossy fiber projection in hippocampus by CRMP2 and CRMP4.

Axon pruning facilitates the removal of ectopic and misguided axons and plays an important role in neural circuit formation during brain development. Sema3F and its receptor neuropilin-2 (Nrp2) have been shown to be involved in the stereotyped pruning of the infrapyramidal bundle (IPB) of mossy fibers of the dentate gyrus (DG) in the developing hippocampus. Collapsin response mediator proteins (CRMPs) were originally identified as an intracellular mediator of semaphorin signaling, and the defective pruning of IPB was recently reported in CRMP2-/- and CRMP3-/- mice. CRMP1 and CRMP4 have high homology to CRMP2 and CRMP3, and their expression in the developing mouse brain overlaps; however, their role in IPB pruning has not yet been examined. In this study, we report that CRMP4, but not CRMP1, is involved in IPB pruning during neural circuit formation in the hippocampus. Our genetic interaction analyses indicated that CRMP2 and CRMP4 have distinct functions and that CRMP2 mediates IPB pruning via Nrp2. We also observed the altered synaptic terminals of mossy fibers in CRMP2 and CRMP4 mutant mice. These results suggest that CRMP family members have a distinct function in the axon pruning and targeting of mossy fibers of the hippocampal DG in the developing mouse brain.

Simple methods for the lesion detection and severity grading of diabetic retinopathy by image processing and transfer learning.

Diabetic retinopathy (DR) has become one of the major causes of blindness. Due to the increased prevalence of diabetes worldwide, diabetic patients exhibit high probabilities of developing DR. There is a need to develop a labor-less computer-aided diagnosis system to support the clinical diagnosis. Here, we attempted to develop simple methods for severity grading and lesion detection from retinal fundus images. We developed a severity grading system for DR by transfer learning with a recent convolutional neural network called EfficientNet-B3 and the publicly available Kaggle Asia Pacific Tele-Ophthalmology Society (APTOS) 2019 training dataset, which includes artificial noise. After removing the blurred and duplicated images from the dataset using a numerical threshold, the trained model achieved specificity and sensitivity values â‰³ 0.98 in the identification of DR retinas. For severity grading, the classification accuracy values of 0.84, 0.95, and 0.98 were recorded for the 1st, 2nd, and 3rd predicted labels, respectively. The utility of EfficientNets-B3 for the severity grading of DR as well as the detailed retinal areas referred were confirmed via visual explanation methods of convolutional neural networks. Lesion extraction was performed by applying an empirically defined threshold value to the enhanced retinal images. Although the extraction of blood vessels and detection of red lesions occurred simultaneously, the red and white lesions, including both soft and hard exudates, were clearly extracted. The detected lesion areas were further confirmed with ground truth using the DIARETDB1 database images with general accuracy. The simple and easily applicable methods proposed in this study will aid in the detection and severity grading of DR, which might help in the selection of appropriate treatment strategies for DR.

Cortical transcriptome analysis after spinal cord injury reveals the regenerative mechanism of central nervous system in CRMP2 knock-in mice.

Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2 (CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function after spinal cord injury (SCI). We aimed to reveal the intracellular mechanism in axotomized neurons in the CRMP2 knock-in (CRMP2KI) mouse model by performing transcriptome analysis in mouse sensorimotor cortex using micro-dissection punching system. Prior to that, we analyzed the structural pathophysiology in axotomized or neighboring neurons after SCI and found that somatic atrophy and dendritic spine reduction in sensorimotor cortex were suppressed in CRMP2KI mice. Further analysis of the transcriptome has aided in the identification of four hemoglobin genes Hba-a1, Hba-a2, Hbb-bs, and Hbb-bt that are significantly upregulated in wild-type mice with concomitant upregulation of genes involved in the oxidative phosphorylation and ribosomal pathways after SCI. However, we observed substantial upregulation in channel activity genes and downregulation of genes regulating vesicles, synaptic function, glial cell differentiation in CRMP2KI mice. Moreover, the transcriptome profile of CRMP2KI mice has been discussed wherein energy metabolism and neuronal pathways were found to be differentially regulated. Our results showed that CRMP2KI mice displayed improved SCI pathophysiology not only via microtubule stabilization in neurons, but also possibly via the whole metabolic system in the central nervous system, response changes in glial cells, and synapses. Taken together, we reveal new insights on SCI pathophysiology and the regenerative mechanism of central nervous system by the inhibition of CRMP2 phosphorylation at Ser522. All these experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee at Waseda University, Japan (2017-A027 approved on March 21, 2017; 2018-A003 approved on March 25, 2018; 2019-A026 approved on March 25, 2019).