A Late Phase of Cerebellar Long-Term Depression Requires Activation of CaMKIV and CREB

AbstractRecently, it has been shown that cerebellar LTD has a late phase that may be blocked by protein synthesis inhibitors. To understand the mechanisms underlying the late phase, we interfered with the activation of transcription factors that might couple synaptic activation to protein synthesis. Particle-mediated transfection of cultured Purkinje neurons with an expression vector encoding a dominant inhibitory form of CREB resulted in a nearly complete blockade of the late phase. Kinases that activate CREB were inhibited, and LTD was assessed. Inhibition of PKA or the MAPK/RSK cascades were without effect on the late phase, while constructs designed to interfere with CaMKIV function attenuated the late phase. These results indicate that the activation of CaMKIV and CREB are necessary to establish a late phase of cerebellar LTD

Sonic hedgehog expressing and responding cells generate neuronal diversity in the medial amygdala

<p>Abstract</p> <p>Background</p> <p>The mammalian amygdala is composed of two primary functional subdivisions, classified according to whether the major output projection of each nucleus is excitatory or inhibitory. The posterior dorsal and ventral subdivisions of the medial amygdala, which primarily contain inhibitory output neurons, modulate specific aspects of innate socio-sexual and aggressive behaviors. However, the development of the neuronal diversity of this complex and important structure remains to be fully elucidated.</p> <p>Results</p> <p>Using a combination of genetic fate-mapping and loss-of-function analyses, we examined the contribution and function of <it>Sonic hedgehog </it>(<it>Shh</it>)-expressing and <it>Shh</it>-responsive (<it>Nkx2-1</it>⁺and <it>Gli1</it>⁺) neurons in the medial amygdala. Specifically, we found that <it>Shh- </it>and <it>Nkx2-1-</it>lineage cells contribute differentially to the dorsal and ventral subdivisions of the postnatal medial amygdala. These <it>Shh</it>- and <it>Nkx2-1</it>-lineage neurons express overlapping and non-overlapping inhibitory neuronal markers, such as Calbindin, FoxP2, nNOS and Somatostatin, revealing diverse fate contributions in discrete medial amygdala nuclear subdivisions. Electrophysiological analysis of the <it>Shh</it>-derived neurons additionally reveals an important functional diversity within this lineage in the medial amygdala. Moreover, inducible <it>Gli1^CreER(T2)</it>temporal fate mapping shows that early-generated progenitors that respond to <it>Shh </it>signaling also contribute to medial amygdala neuronal diversity. Lastly, analysis of <it>Nkx2-1 </it>mutant mice demonstrates a genetic requirement for <it>Nkx2-1 </it>in inhibitory neuronal specification in the medial amygdala distinct from the requirement for <it>Nkx2-1 </it>in cerebral cortical development.</p> <p>Conclusions</p> <p>Taken together, these data reveal a differential contribution of <it>Shh-</it>expressing and <it>Shh</it>-responding cells to medial amygdala neuronal diversity as well as the function of <it>Nkx2-1 </it>in the development of this important limbic system structure.</p

Gli3 Repressor Controls Cell Fates and Cell Adhesion for Proper Establishment of Neurogenic Niche

Neural stem cells (NSCs) in the subventricular zone (SVZ) rely on environmental signals provided by the neurogenic niche for their proper function. However, little is known about the initial steps of niche establishment, as embryonic radial glia transition to postnatal NSCs. Here, we identify Gli3 repressor (Gli3R), a component of the Sonic hedgehog (Shh) pathway, as a critical factor controlling both cell-type specification and structural organization of the developing SVZ. We demonstrate that Gli3R expressed in radial glia temporally regulates gp130/STAT3 signaling at the transcriptional level to suppress glial characteristics in differentiating ependymal cells. In addition, Gli3R maintains the proper level of Numb in ependymal cells to allow localization of cell adhesion molecules such as vascular cell adhesion molecule (VCAM) and E-cadherin. Thus, our findings reveal a role for Gli3R as a mediator of niche establishment and provide insights into the conditions required for proper SVZ neurogenic niche formation

Exosome Proteome of U-87MG Glioblastoma Cells

Exosomes are small membrane vesicles between 30 and 100 nm in diameter secreted by many cell types, and are associated with a wide range of physiological and/or pathological processes. Exosomes containing proteins, lipids, mRNA, and microRNA contribute to cell-to-cell communication and cell-to-environment regulation, however, their biological functions are not yet fully understood. In this report, exosomes in the glioblastoma cell line, U-87MG, were isolated and the proteome was investigated. In addition, exosome proteome changes in U-87MG cells exposed to a low temperature were investigated to elucidate whether the exosome proteome could respond to an external stimulus. Cell culture medium was collected, and exosomes were isolated by continuous centrifugation eliminating cell debris, nucleic acids, and other particles. The morphology of exosomes was observed by cryo-tunneling electron microscopy. According to 2-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, certain proteins including collagen type VI alpha 1, putative RNA-binding protein 15B chain A, substrate induced remodeling of the active site regulates HTRA1, coatomer protein complex-subunit beta 2, myosin-heavy chain 1, and keratin-type I cytoskeletal 9 showed differences between the control proteome and the low temperature-exposed proteome

One-Step Gait Pattern Analysis of Hip Osteoarthritis Patients Based on Dynamic Time Warping through Ground Reaction Force

Osteoarthritis (OA) of the hip is a degenerative joint disease, which means it causes gradual damage to the joint, and its incidence rate continues to increase worldwide. Degenerative osteoarthritis can cause significant pain and gait disturbance in walking, affecting daily life. A diagnosis method for hip OA includes questioning and various walking movements to find abnormalities of gait patterns based on human observation. However, when multiple gait tests are performed to notice the gait, it can cause pain continuously, even during the examination. Suppose hip OA could be diagnosed with only a one-step gait; both patients and medical doctors would be benefited because the diagnosis time can be reduced and the burden on the patient is decreased dramatically. Therefore, in this paper, we aimed to propose a method to recognize the abnormality of the hip OA patient with a one-step gait pattern based on a dynamic time warping (DTW) algorithm through three directional ground reaction forces (GRFs). After a force plate measured three directional GRFs, the data of twenty-three hip OA patients and eighteen healthy people were classified using supervised machine learning algorithms. The results of the classification showed high accuracy and reliability. Then, the DTW algorithm was applied to compare the data of patients and healthy people to find out when patients may feel pain during the gait. By applying the DTW algorithm, it was possible to find out in which gait phase the patient’s gait showed the difference, such as when the heel first contacted the ground, in the middle of walking, or when the toe came off the ground. Through the results, the data of the one-step gait on the force plate enabled us to classify patients and healthy people with a high accuracy of over 70%, recognize the abnormal gait pattern, and determine how to relieve the pain during the gait

Development of Inclined Applicators for Flattened Beam Intraoperative Radiotherapy

A 3D-printed applicator for intraoperative radiation therapy (IORT) using the INTRABEAM system has been introduced, designed to protect normal tissue, expand indications, and assess its effectiveness in various intraoperative settings. The study involved designing and creating inclined applicators at 0° and 45° angles, which were capable of producing flattened and unflattened beams and shielding non-target areas from radiation. The applicator was evaluated based on dose uniformity, percentage depth dose, leakage, and dose rate. The findings showed that the applicator provided a uniform beam at various depths (1.23 to 1.30 for the flattened beam and 1.25 to 1.32 for the unflattened beam) and effectively shielded against leakage, with no leakage dose measured at 1 mm from the surface. Additionally, the dose rates after 3 min of irradiation for flattened and unflattened beams were 51.0 and 52.9 cGy for 0° and 48.5 and 54.8 cGy for 45° applicators, respectively. This suggests that the applicator can efficiently deliver IORT while minimizing exposure to normal tissues, particularly in treating chest and abdominal lesions near or on critical organs. The applicator has undergone dose calibration and has moved forward to clinical application