Imaging calcium microdomains within entire astrocyte territories and endfeet with GCaMPs expressed using adeno-associated viruses.
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...It is important to note that the Ca transients that occur in astrocytes are of a much slower timescale than what occurs in neurons during neurotransmission (Shigetomi et al., 2016)....
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...It is important to note that the Ca2+ transients that occur in astrocytes are of a much slower timescale than what occurs in neurons during neurotransmission (Shigetomi et al., 2016)....
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"Imaging calcium microdomains within..." refers methods in this paper
...For membrane targeting in situ after in vivo expression, we used the N-terminal domain of Lck, a Src tyrosine kinase, that efficiently recruits proteins to the membrane (Shigetomi et al., 2010b, 2012) (Fig....
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...…(AAV)2/5 generation Plasmids encoding cytosolic GCaMP3 (cyto-GCaMP3) and membrane-targeted Lck-GCaMP3 were described previously (Tian et al., 2009; Shigetomi et al., 2012) (Addgene plasmids 22692 and 26974, Shigetomi et al. 635 were washed three times with PBS before being mounted on glass…...
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"Imaging calcium microdomains within..." refers background in this paper
...Moreover, the ability to measure endfeet Ca signals directly and simultaneously with other parts of the astrocyte will aid the exploration of neurovascular coupling mechanisms in health and disease (Attwell et al., 2010)....
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...Moreover, the ability to measure endfeet Ca2+ signals directly and simultaneously with other parts of the astrocyte will aid the exploration of neurovascular coupling mechanisms in health and disease (Attwell et al., 2010)....
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...It is now well established that astrocytes serve vital physiological roles in the functioning of the nervous system, including buffering of K around neurons, regulation of blood flow, clearance of neurotransmitters from synapses, as well as providing trophic factors and nutrients (Kofuji and Newman, 2004; Barres, 2008; Attwell et al., 2010)....
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...In addition to these vital contributions to the functioning of the brain as a tissue, evidence suggests that astrocytes may respond to, and regulate, neuronal function and blood flow (Araque et al., 2001; Haydon, 2001; Attwell et al., 2010; Halassa and Haydon, 2010)....
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...…vital physiological roles in the functioning of the nervous system, including buffering of K+ around neurons, regulation of blood flow, clearance of neurotransmitters from synapses, as well as providing trophic factors and nutrients (Kofuji and Newman, 2004; Barres, 2008; Attwell et al., 2010)....
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1,862 citations
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...Additionally, patch-mediated loading of dyes is known to dialyze and disrupt astrocyte functions (Nett et al., 2002) and can only be performed on cells one-by-one, whereas bulk loading results in uneven loading and is troublesome in adult tissue (Kang and Nedergaard, 2000; Tong et al., 2012)....
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...As recently discussed, we also considered the higher fluorescence of GCaMP3 desirable for imaging the highly ramified astrocytes in thick brain slices, which scatter light (Tong et al., 2012)....
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...…resonance energy transfer–based calcium sensor expressed in astrocytes in vivo (Atkin et al., 2009; Russell, 2011), and recently progress has been made using single-wavelength GECIs to study Ca2+ signals in astrocytes (Shigetomi et al., 2010a,b, 2012; Arizono et al., 2012; Tong et al., 2012)....
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...…cell physiology (Rand et al., 1994), and the high concentrations of Ca2+ indicator dyes often used to see Ca2+ signals in astrocyte processes (see Tong et al., 2012) are not Imaging calcium microdomains within entire astrocyte territories and endfeet with GCaMPs expressed using adeno-associated…...
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...…of the aforementioned problems may be addressed with the use of genetically encoded Ca2+ indicators (GECIs), which could be expressed in multiple cells without the need for dialyzing astrocytes with high concentrations of organic dyes via patch pipettes, as recently discussed (Tong et al., 2012)....
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1,385 citations
"Imaging calcium microdomains within..." refers background in this paper
...In addition to these vital contributions to the functioning of the brain as a tissue, evidence suggests that astrocytes may respond to, and regulate, neuronal function and blood flow (Araque et al., 2001; Haydon, 2001; Attwell et al., 2010; Halassa and Haydon, 2010)....
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