We are using a coination of parasite cell biology, live cell imaging and single molecule biophysics to elucidate the molecular mechanisms underlying dense granule transport and secretion. By understanding the mechanisms underlying this essential process it is our goal to identify new targets for the development of anti-parasitic drugs.
Title: Functional Evaluation of Neural Stem Cell Differentiation by Single Cell Calcium Imaging VOLUME: 6 ISSUE: 3 Author(s):Maria Francisca Eiriz, Sofia Grade, Alexandra Rosa, Sara Xapelli, Liliana Bernardino, Fabienne Agasse and Joao O. Malva Affiliation:Neuroprotection and Neurogenesis in Brain Repair Group, Center for Neuroscience and Cell Biology, Institute of Biochemistry, Faculty of
In a calcium imaging study the PGD 2 metabolite 15-deoxy-Δ 12,14-prostaglandin J 2 activated the human TRPA1 expressed in HEK293 cells and in a subset of chemosensitive mouse trigeminal neurons, where this activation was blocked by both the nonselective TRP channel blocker ruthenium red, and the TRPA1 inhibitor HC-030031.
Our recent paper on functional characterisation of granule cells has been published in the journal Current Biology, and we are following up on this research by examining other cell types, representations of this behavior in retinorecipient areas and in central brain regions using two-photon calcium imaging.
Oct 16, 2017· Intracellular calcium is an important ion involved in the regulation and modulation of many neuronal functions. From regulating cell cycle and proliferation to initiating signaling cascades and regulating presynaptic neurotransmitter release, the concentration and timing of calcium activity governs the function and fate of neurons. Changes in calcium transients can be used in high-throughput
Mar 13, 2016· “Here, we were able to demonstrate that adult-born granule cells act differently than their mature neighbors, and determine why that difference is so critical.” Using calcium imaging and a miniature microscope implanted into the brains of live mice, the researchers found that new neurons exhibited a burst of excitability after genesis.
Ca 2+ transients were recorded in Oregon-Green-BAPTA 1-loaded brain slices using a calcium imaging technique. For the detection, identifiion and characterisation of the Ca 2+ transients, a wavelet analysis-based method was developed. Granule cells were …
To address this question, we placed a stimulating electrode in the granule cell layer to activate centrifugal glutamatergic inputs that synapse onto granule cell dendrites (Balu et al. 2007). We delivered a train of stimuli (10 Hz, 30 s) to this pathway and used calcium imaging to monitor the activity of NPCs in the adjacent SEL of the
Labeled calcium indiors are molecules that exhibit an increase in fluorescence upon binding Ca 2+.They have uses in many calcium signaling investigations, including measuring intracellular Ca 2+, following Ca 2+ influx and release, and multiphoton excitation imaging of Ca 2+ in living tissues. Cells may be loaded with the AM ester forms of these calcium indiors by adding the dissolved
Calcium imaging is an extremely useful technique for investigating the variety of roles that calcium ions have in functioning neurons. Calcium ions generate a multitude of intracellular signals that control key functions, such as neurotransmitter release from synaptic vesicles.
Further electrophysiology and calcium imaging analyses demonstrated the maturation, connectivity and network properties of these cells in the astrocyte co-culture system (Fig. 5B). The isolated NPC population was also successfully transplanted into the developing DG of P10 animals to produce electrophysiologically active Prox1 + neurons.
Sep 25, 2017· Two-photon calcium imaging reveals the in vivo activity of multiple neurons at cellular and subcellular resolution (Jia et al., 2010; Ohki et al., 2005).Recent work demonstrates that by exciting red-fluorescent calcium indiors with a laser at wavelengths of 1000–1100 nm through a cranial window, it is possible to image neural activity in the mouse sensory cortex at depths of 800–900 µm
Apr 23, 2019· Two-photon calcium imaging is a standard technique of neuroscience laboratories that records neural activity from individual neurons over large populations in awake-behaving animals. Automatic and accurate identifiion of behaviorally relevant neurons from these recordings is a critical step toward complete mapping of brain activity. To this end, we present a fast deep learning …
May 16, 2016· Calcium-imaging is a sensitive method for monitoring calcium dynamics during neuronal activity. As intracellular calcium concentration is correlated to physiological and pathophysiological activity of neurons, calcium imaging with fluorescent indiors is one of the most commonly used techniques in neuroscience today. Current methodologies for loading calcium dyes …
Because each system or cell type has a different calcium signaling tool kit, a GECI with high sensitivity in one setting might not be the best fit when conditions change. The GECI performance in vitro correlates poorly with that in more intact preparations (Tian et al. 2009). Therefore, we sought to establish a standardized, multistep screening
of calcium imaging datasets, which includes non-gaussian noise, non-cell background activity (e.g. neuropil), and overlapping cells not captured by algorithms (out-of-focus or foreground). As a consequence, the impact of such impurities inherent in calcium imaging on the accuracy of extracted signals has not been thoroughly investigated previously.
The laboratory research is both experimental and computational, with strong emphasis on optical brain imaging and machine learning applied to data analysis. The NEL lab develops tools for simultaneous recording, modulation and real time extraction of neural dynamics at different brain loions and scales.
May 06, 2019· Tu MK, Borodinsky LN. (2014) Spontaneous calcium transients manifest in the regenerating muscle and are necessary for skeletal muscle replenishment. Cell Calcium, 56: 34-41. Spitzer NC, Borodinsky LN, Root CM. (2013) Imaging and manipulating calcium transients in developing Xenopus spinal neurons. Cold Spring Harbor Protocols. doi:10.1101/pdb
gyrus granule cells (GCs) generated postnatally (Lledo et al. 2006). In rodents, it was shown that the synaptic integration and survival of the young GCs is strongly dependent on the experience of the animal during a Calcium imaging Dendritic Ca2+ signals were recorded either on proximal
Mar 02, 2009· However, the cells would need to be re-stained with a calcium indior dye (by inserting a micropipette into the brain) each time before imaging.” She noted that a recent study published in Nature Methods ( Mank et al., 2008 ) described the advance of a genetically encoded calcium indior TN-XXL, which allowed calcium increases in neurons
January: Our paper on in vivo characterization of hippocampal mossy cells with calcium imaging is published in Neuron. our collaborative paper with the Hen lab on in vivo imaging dentate gyrus granule cells was selected for the Best of Neuron 2015-16; 2015. February: Congratulation to Jeff Zarea on his NIH NRSA Predoctoral Fellowship!
Other major interests of the group are the mechanisms of the input integration in granule cells (#5 published in 2016, #6 2013). To directly measure the propagation of signals along the dendritic arbors of granule cells we employ dendritic recordings, optical stimulation and calcium imaging, and verifiion of the obtained data by
To monitor depolarization-induced intracellular calcium increases in neurons, cells expressing the genetically encoded calcium reporter YC3.60 (Nagai et al., 2004, Cao et al., 2005) were washed with calcium-containing Locke’s buffer either manually or using the Wellwash Versa, and kept in a 100 μl volume before performing live cell imaging.
In a mouse model of temporal lobe epilepsy, multicellular calcium imaging revealed that disease emergence was accompanied by massive amplifiion in the normally sparse, afferent stimulation-induced activation of hippocampal dentate granule cells. Patch recordings demonstrated reductions in local inhibitory function within the dentate gyrus at time points where sparse activation was …
Calcium Imagingbutton . A new tab Calcium Imaging appears with two formu-las for calcium calculations. The upper formula is used for calculation of the Fluorescence Intensity Ratio. R = F (340 nm)/ F (380 nm) The second formula is used for evaluation of the Calcium Concentration according to Grynkiewicz (see Reference).
Cell 2019 Dec 12;179(7):1590-1608.e23. doi: 10.1016/j.cell.2019.11.004. Y. Mircheva, M. R. Peralta III and K. Tóth (2019) Recruitment of feedforward inhibition in the molecular layer of the dentate gyrus drives long lasting hyperpolarization in granule cells and alters the entorhinal input integration. J Neurosci.