A novel, biophysically realistic model for early-stage, acetylcholine-mediated retinal waves is presented. In this model, neural excitability is regulated through a slow after-hyperpolarization (sAHP) operating on two different temporal scales. As a result, the simulated network exhibits competition between a desynchronizing effect of spontaneous, cell-intrinsic bursts, and the synchronizing effect of synaptic transmission during retinal waves. Cell-intrinsic bursts decouple the retinal network through activation of the sAHP current, and we show that the network is capable of operating at a transition point between purely local and global functional connectedness, which corresponds to a percolation phase transition. Multielectrode array recordings show that, at this point, the properties of retinal waves are reliably predicted by the model. These results indicate that early spontaneous activity in the developing retina is regulated according to a very specific principle, which maximizes randomness and variability in the resulting activity patterns.
We have investigated the use of carbon nanotube coated microelectrodes as an interface material for retinal recording and stimulation applications. Test devices were micro-fabricated and consisted of 60, 30 μm diameter electrodes at spacing of 200 μm. These electrodes were coated via chemical vapor deposition of carbon nanotubes, resulting in conducting, three dimensional surfaces with a high interfacial area. These attributes are important both for the quality of the cell-surface coupling as well as for electro-chemical interfacing efficiency. The entire chip was packaged to fit a commercial multielectrode recording and stimulation system. Electrical recordings of spontaneous spikes from whole-mount neonatal mouse retinas were consistently obtained minutes after retinas were placed over the electrodes, exhibiting typical bursting and propagating waves. Most importantly, the signals obtained with carbon nanotube electrodes have exceptionally high signal to noise ratio, reaching values as high as 75. Moreover, spikes are marked by a conspicuous gradual increase in amplitude recorded over a period of minutes to hours, suggesting improvement in cell-electrode coupling. This phenomenon is not observed in conventional commercial electrodes. Electrical stimulation using carbon nanotube electrodes was also achieved. We attribute the superior performances of the carbon nanotube electrodes to their three dimensional nature and the strong neuro-carbon nanotube affinity. The results presented here show the great potential of carbon nanotube electrodes for retinal interfacing applications. Specifically, our results demonstrate a route to achieve a reduction of the electrode down to few micrometers in order to achieve high efficacy local stimulation needed in retinal prosthetic devices.
Abstract. To study the mechanisms responsible for the regulation of flagellar length, we examined the effects of colchicine and Cytochalasin D (CD) on the growth and maintenance of Chlamydomonas flagella on motile wild type cells as well as on pf 18 cells, whose flagella lack the centrM microtubules and are immobile. CD had no effect on the regeneration of flagella after deflagellation but it induced fully assembled flagella to shorten at an average rate of 0.03 t~m-min I. Cells remained fully motile in CD and even stubby flagella continued to move, indicating that flagellar shortening did not selectively disrupt machinery necessary for motility. To observe the effects of the drug on individual cells, pf 18 cells were treated with CD and flagella on cells were monitored by direct observation over a 5-hour period. Flagella on control pf 18 cells maintained their initial lengths throughout the experiment but flagella on CD-treated cells exhibited periods of elongation, shortening, and regrowth suggestive of the dynamic behavior of cytoplasmic microtubules observed in vitro and in vitro. Cells behaved individually, with no two cells exhibiting the same flageltar behavior at any given time although both flagella on any single cell behaved identically.The rate of drug-induced flagellar shortening and elongation in pf 18 cells varied from 0.08 to 0.17 #m-min -t, with each event occurring over 10-60-min periods. Addition of colchicine to wiM type and pf 18 cells induced flagella to shorten at an average rate of 0.06 #m-min -~ until the flagella reached an average of 73% of their initial length, after which they exhibited no further shortening or elongation. Cells treated with colchicine and CD exhibited nearly complete flagellar resorption, with little variation in flagellar length among cells. The effects of these drugs were reversible and flagella grew to normal stable lengths after drug removal. Taken together, these results show that the distal half to one-third of the Chlamydomonas flagellum is relatively unstable in the presence of colchicine but that the proximal half to two-thirds of the flagellum is stable to this drug. In contrast to colchicine, CD can induce nearly complete flagellar microtubule disassembly as well as flagellar assembly. Flagellar microtubules must, therefore, be inherently unstable, and flagellar length is stabilized by factors that are sensitive, either directly or indirectly, to the effects of CD.
Fluorescent nanodiamonds (fNDs) containing nitrogen vacancy (NV) centers are promising candidates for quantum sensing in biological environments. This work describes the fabrication and implementation of electrospun poly lactic‐co‐glycolic acid (PLGA) nanofibers embedded with fNDs for optical quantum sensing in an environment, which recapitulates the nanoscale architecture and topography of the cell niche. A protocol that produces uniformly dispersed fNDs within electrospun nanofibers is demonstrated and the resulting fibers are characterized using fluorescent microscopy and scanning electron microscopy (SEM). Optically detected magnetic resonance (ODMR) and longitudinal spin relaxometry results for fNDs and embedded fNDs are compared. A new approach for fast detection of time varying magnetic fields external to the fND embedded nanofibers is demonstrated. ODMR spectra are successfully acquired from a culture of live differentiated neural stem cells functioning as a connected neural network grown on fND embedded nanofibers. This work advances the current state of the art in quantum sensing by providing a versatile sensing platform that can be tailored to produce physiological‐like cell niches to replicate biologically relevant growth environments and fast measurement protocols for the detection of co‐ordinated endogenous signals from clinically relevant populations of electrically active neuronal circuits.
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