Super-resolution 2-photon microscopy reveals that the morphology of each dendritic spine correlates with diffusive but not synaptic properties

The structure of dendritic spines suggests a specialized function in compartmentalizing synaptic signals near active synapses. Indeed, theoretical and experimental analyses indicate that the diffusive resistance of the spine neck is sufficient to effectively compartmentalize some signaling molecules in a spine for the duration of their activated lifetime. Here we describe the application of 2-photon microscopy combined with stimulated emission depletion (STED-2P) to the biophysical study of the relationship between synaptic signals and spine morphology, demonstrating the utility of combining STED-2P with modern optical and electrophysiological techniques. Morphological determinants of fluorescence recovery time were identified and evaluated within the context of a simple compartmental model describing diffusive transfer between spine and dendrite. Correlations between the neck geometry and the amplitude of synaptic potentials and calcium transients evoked by 2-photon glutamate uncaging were also investigated

Orbital and meteorological factors pertinent to satellite transmissions of facsimile weather charts Final report

Automatic picture transmission system on Nimbus and earth synchronous satellites for transmission of weather chart

Treatment of Congenital Myogenic Ptosis with Frontalis Suspension Surgery

Ptosis repair is a challenging oculoplastic surgical procedure that requires correct diagnosis, thoughtful planning, thorough understanding of eyelid anatomy, experience, and good surgical technique. Ptosis can causefunctional problem and cosmetic issue.Thisstudywasreport 2 cases of congenital myogenic ptosis which were treated with frontalis suspension surgery.Case report,a 11-year old and a 20-year old girl came to our clinic with chief complain drooping in the upper eyelid RE since their childhood. Patients also complained difficulty of reading. The degree of ptosis during the day was the same. No diplopia, dysphonia, dyspnea, and dysphagia. VA 20/20 in the both eyes. Ocular motility were normal. MRD1 0 mm, FP 5 mm, no upper eyelid crease, LF 4 mm, no lagophthalmos, eyelid lag present, upper eyelid RE higher than LE in down gaze, and Bells phenomenon normal. Light reflex of pupil positive and diameter 3 mm in standard room illumination. Schimer test and TBUT were within normal. We diagnosed these patients with congenital myogenic ptosis RE and performed frontalis suspension surgery with autogenous tensor fascia lata.Postoperative, for first patient we got MRD1 2 mm and FP 7 mm. For second patient we got MRD14 mm and FP 9 mm. Patients had no complain drooping in the upper eyelid RE and difficulty of reading anymore.Frontalis suspension surgery is a way to treat patient with congenital myogenic ptosis who have poor levator function

A PKA activity sensor for quantitative analysis of endogenous GPCR signaling via 2-photon FRET-FLIM imaging

Neuromodulators have profound effects on behavior, but the dynamics of their intracellular effectors has remained unclear. Most neuromodulators exert their function via G-protein-coupled receptors (GPCRs). One major challenge for understanding neuromodulator action is the lack of dynamic readouts of the biochemical signals produced by GPCR activation. The adenylate cyclase/cyclic AMP/protein kinase A (PKA) module is a central component of such biochemical signaling. This module is regulated by several behaviorally important neuromodulator receptors. Furthermore, PKA activity is necessary for the induction of many forms of synaptic plasticity as well as for the formation of long-term memory. In order to monitor PKA activity in brain tissue, we have developed a 2-photon fluorescence lifetime imaging microscopy (2pFLIM) compatible PKA sensor termed FLIM-AKAR, which is based on the ratiometric FRET sensor AKAR3. FLIM-AKAR shows a large dynamic range and little pH sensitivity. In addition, it is a rapidly diffusible cytoplasmic protein that specifically reports net PKA activity in situ. FLIM-AKAR expresses robustly in various brain regions with multiple transfection methods, can be targeted to genetically identified cell types, and responds to activation of both endogenous GPCRs and spatial-temporally specific delivery of glutamate. Initial experiments reveal differential regulation of PKA activity across subcellular compartments in response to neuromodulator inputs. Therefore, the reporter FLIM-AKAR, coupled with 2pFLIM, enables the study of PKA activity in response to neuromodulator inputs in genetically identified neurons in the brain, and sheds light on the intracellular dynamics of endogenous GPCR activation

CRISPR/Cas9-Mediated Gene Knock-Down in Post-Mitotic Neurons

The prokaryotic adaptive immune system CRISPR/Cas9 has recently been adapted for genome editing in eukaryotic cells. This technique allows for sequence-specific induction of double-strand breaks in genomic DNA of individual cells, effectively resulting in knock-out of targeted genes. It thus promises to be an ideal candidate for application in neuroscience where constitutive genetic modifications are frequently either lethal or ineffective due to adaptive changes of the brain. Here we use CRISPR/Cas9 to knock-out Grin1, the gene encoding the obligatory NMDA receptor subunit protein GluN1, in a sparse population of mouse pyramidal neurons. Within this genetically mosaic tissue, manipulated cells lack synaptic current mediated by NMDA-type glutamate receptors consistent with complete knock-out of the targeted gene. Our results show the first proof-of-principle demonstration of CRISPR/Cas9-mediated knock-down in neurons in vivo, where it can be a useful tool to study the function of specific proteins in neuronal circuits

Emergence of Synchronous Oscillations in Neural Networks Excited by Noise

The presence of noise in non linear dynamical systems can play a constructive role, increasing the degree of order and coherence or evoking improvements in the performance of the system. An example of this positive influence in a biological system is the impulse transmission in neurons and the synchronization of a neural network. Integrating numerically the Fokker-Planck equation we show a self-induced synchronized oscillation. Such an oscillatory state appears in a neural network coupled with a feedback term, when this system is excited by noise and the noise strength is within a certain range.Comment: 12 pages, 18 figure

A Direct Projection from Mouse Primary Visual Cortex to Dorsomedial Striatum

The mammalian striatum receives inputs from many cortical areas, but the existence of a direct axonal projection from the primary visual cortex (V1) is controversial. In this study we use anterograde and retrograde tracing techniques to demonstrate that V1 directly innervates a topographically defined longitudinal strip of dorsomedial striatum in mice. We find that this projection forms functional excitatory synapses with direct and indirect pathway striatal projection neurons (SPNs) and engages feed-forward inhibition onto these cells. Importantly, stimulation of V1 afferents is sufficient to evoke phasic firing in SPNs. These findings therefore identify a striatal region that is functionally innervated by V1 and suggest that early visual processing may play an important role in striatal-based behaviors