What is gcamp6s?
Summary. Fluorescent calcium sensors are widely used to image neural activity. Using structure-based mutagenesis and neuron-based screening, we developed a family of ultra-sensitive protein calcium sensors (GCaMP6) that outperformed other sensors in cultured neurons and in zebrafish, flies, and mice in vivo.
What are genetically encoded calcium indicators?
The genetically encoded calcium indicator (GECI), which binds to calcium ions and emits fluorescence visualizing intracellular calcium concentration, enables detection of in vivo neuronal firing activity. Various GECIs have been developed and can be chosen for diverse purposes.
Who invented GCaMP?
Junichi Nakai
GCaMP is a genetically encoded calcium indicator (GECI) initially developed in 2001 by Junichi Nakai. It is a synthetic fusion of green fluorescent protein (GFP), calmodulin (CaM), and M13, a peptide sequence from myosin light-chain kinase.
What is in vivo fiber photometry?
Fiber photometry is an in vivo calcium imaging method that detects average fluorescence intensity changes. Thus, this method is used to measure population neural activity in a freely-behaving animal (Cui et al. 2014).
What is neuronal calcium imaging?
Calcium imaging enables researchers to investigate the highly synchronous network activity of neurons. Here, neuroscientists used calcium imaging to assess the fluorescent signal of 40 neurons. With this information, network properties such as signal propagation and neural correlations can be determined.
Why are calcium ions important for signaling?
Calcium signaling is the use of calcium ions (Ca2+) to communicate and drive intracellular processes often as a step in signal transduction. Ca2+ is important for cellular signalling, for once it enters the cytosol of the cytoplasm it exerts allosteric regulatory effects on many enzymes and proteins.
Why is calcium imaging important?
Calcium ions generate versatile intracellular signals that control key functions in all types of neurons. Imaging calcium in neurons is particularly important because calcium signals exert their highly specific functions in well-defined cellular subcompartments.
What is 2 photon calcium imaging?
Two-photon calcium imaging is a powerful means for monitoring the activity of distinct neurons in brain tissue in vivo. In the mammalian brain, such imaging studies have been restricted largely to calcium recordings from neurons that were individually dye-loaded through microelectrodes.
What does GCaMP measure?
GECIs measure calcium, the key signal molecule underlying cell functions such as heart, vessel, and airway contraction, lung secretion, autonomic neurotransmission, and immunocyte function. GCaMP, originally developed by Junichi Nakai is an example of a GECI.
Is fiber a photometry imaging?
Fiber photometry is an imaging method that enables scientists to image population-level neural activity in the brain of freely-behaving animals.
How does fiber photometry work?
Fiber photometry, a technique related to optogenetics, provides detailed insight into the activity and behavior of neuronal populations. This technique stimulates neurons with light and measures fluorescence signals that correspond to calcium dynamics.
Which gcamp5s are expressed in larval Drosophila?
Characterization of GCaMP5s in larval Drosophila Both GCaMP3 and 5G were expressed in Drosophila melanogasterlarvae using a motor-neuron promoter (OK6-Gal4). Type 1b NMJ boutons of third instar larvae were imaged following activity evoked from electrically stimulating motor neuron axons using a suction electrode (Macleod et al., 2002) (Fig. 6A).
What is the GCaMP6 family?
The GCaMP6 family of GECIs is a collection of ultrasensitive, green fluorescent indicator proteins that enable reliable detection of single action potential responses in vivo and facilitate the measurement of synaptic calcium signals.
What is the GCaMP protein?
The GCaMP protein is a fusion of the calmodulin-binding domain from the myosin light chain kinase (which is called an M13 peptide), permutated EGFP, and the calmodulin and increases its intensity of green fluorescence upon binding to calcium ions (Nakai, Ohkura, & Imoto, 2001).
What are GCaMP6 indicators?
GCaMP6 indicators cross important performance thresholds. They have higher sensitivity than commonly used synthetic calcium dyes (e.g., OGB1) and detect individual action potentials with high reliability at reasonable microscope magnifications.