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Functional dissection of synaptic circuits: in vivo patch-clamp recording in neuroscience.

Tao C, Zhang G, Xiong Y, Zhou Y - Front Neural Circuits (2015)

Bottom Line: With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals.This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions.The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Chongqing Key Laboratory of Neurobiology, Third Military Medical University Chongqing, China.

ABSTRACT
Neuronal activity is dominated by synaptic inputs from excitatory or inhibitory neural circuits. With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals. This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions. Combining in vivo patch-clamp recording with other techniques, such as two-photon imaging or optogenetics, can provide even clearer functional dissection of the synaptic contributions of different neurons or nuclei. Here, we summarized current applications and recent research progress using the in vivo patch-clamp recording method and focused on its role in the functional dissection of different synaptic inputs. The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed.

No MeSH data available.


Related in: MedlinePlus

Combination of in vivo patch-clamp recording and other techniques. (A) Two-photon guided patch clamp can target specific neurons using fluorescent guidance. RFP, red fluorescent protein. Alexa 488, a green fluorescent dye widely used for two-photon imaging. (B) Optogenetic manipulation of neural circuits can isolate different sources of excitation. Blue light can activate ChR2-expressing PV+ inhibitory neurons in the visual cortex (red neurons) and silence cortical excitatory neurons. Then, the excitatory contribution from the thalamus only (thalamic EPSCs) can be isolated from the mixed input (thalamic + intracortical). Modified from Lien and Scanziani (2013), with permission. (C) Current-clamp and voltage-clamp recordings made from the same neuron in a living mouse. Current-clamp mode can record membrane potential changes and voltage-clamp mode can dissect synaptic currents into excitatory and inhibitory components by holding the cell membrane potential at different levels. The synaptic contributions to orientation selectivity can then be compared and quantified. The right panel depicts the orientation tuning curve for excitatory input (Ex), inhibitory input (In) and membrane potential (Vm). Error bar = SEM. The tuning width (delta) is denoted in the inset. *p < 0.01, paired t-test. Modified from Liu et al. (2011), with permission.
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Figure 2: Combination of in vivo patch-clamp recording and other techniques. (A) Two-photon guided patch clamp can target specific neurons using fluorescent guidance. RFP, red fluorescent protein. Alexa 488, a green fluorescent dye widely used for two-photon imaging. (B) Optogenetic manipulation of neural circuits can isolate different sources of excitation. Blue light can activate ChR2-expressing PV+ inhibitory neurons in the visual cortex (red neurons) and silence cortical excitatory neurons. Then, the excitatory contribution from the thalamus only (thalamic EPSCs) can be isolated from the mixed input (thalamic + intracortical). Modified from Lien and Scanziani (2013), with permission. (C) Current-clamp and voltage-clamp recordings made from the same neuron in a living mouse. Current-clamp mode can record membrane potential changes and voltage-clamp mode can dissect synaptic currents into excitatory and inhibitory components by holding the cell membrane potential at different levels. The synaptic contributions to orientation selectivity can then be compared and quantified. The right panel depicts the orientation tuning curve for excitatory input (Ex), inhibitory input (In) and membrane potential (Vm). Error bar = SEM. The tuning width (delta) is denoted in the inset. *p < 0.01, paired t-test. Modified from Liu et al. (2011), with permission.

Mentions: Compared with the blind-patch procedure, cell location and morphology can be visualized during the recording session in visually guided patch-clamp, which is very useful for recording from specific neurons (e.g., sparsely distributed inhibitory interneurons). To visualize a target neuron in vivo, the cell needs to be either brightened or shadowed. The brightening method uses transgenic or viral methods to visualize the neurons by adding fluorescent protein to the cell membrane (Trachtenberg et al., 2002; Komai et al., 2006; Häusser and Margrie, 2014; Li et al., 2014a). Then, a pipette filled with fluorescent dye can be used guide the hunt for the target neuron (Figure 2A). In the shadowing method, the extracellular matrix surrounding the target region is perfused with a fluorescent dye and brightened; thus, target neurons can be visualized by negative signals (Kitamura et al., 2008).


Functional dissection of synaptic circuits: in vivo patch-clamp recording in neuroscience.

Tao C, Zhang G, Xiong Y, Zhou Y - Front Neural Circuits (2015)

Combination of in vivo patch-clamp recording and other techniques. (A) Two-photon guided patch clamp can target specific neurons using fluorescent guidance. RFP, red fluorescent protein. Alexa 488, a green fluorescent dye widely used for two-photon imaging. (B) Optogenetic manipulation of neural circuits can isolate different sources of excitation. Blue light can activate ChR2-expressing PV+ inhibitory neurons in the visual cortex (red neurons) and silence cortical excitatory neurons. Then, the excitatory contribution from the thalamus only (thalamic EPSCs) can be isolated from the mixed input (thalamic + intracortical). Modified from Lien and Scanziani (2013), with permission. (C) Current-clamp and voltage-clamp recordings made from the same neuron in a living mouse. Current-clamp mode can record membrane potential changes and voltage-clamp mode can dissect synaptic currents into excitatory and inhibitory components by holding the cell membrane potential at different levels. The synaptic contributions to orientation selectivity can then be compared and quantified. The right panel depicts the orientation tuning curve for excitatory input (Ex), inhibitory input (In) and membrane potential (Vm). Error bar = SEM. The tuning width (delta) is denoted in the inset. *p < 0.01, paired t-test. Modified from Liu et al. (2011), with permission.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440909&req=5

Figure 2: Combination of in vivo patch-clamp recording and other techniques. (A) Two-photon guided patch clamp can target specific neurons using fluorescent guidance. RFP, red fluorescent protein. Alexa 488, a green fluorescent dye widely used for two-photon imaging. (B) Optogenetic manipulation of neural circuits can isolate different sources of excitation. Blue light can activate ChR2-expressing PV+ inhibitory neurons in the visual cortex (red neurons) and silence cortical excitatory neurons. Then, the excitatory contribution from the thalamus only (thalamic EPSCs) can be isolated from the mixed input (thalamic + intracortical). Modified from Lien and Scanziani (2013), with permission. (C) Current-clamp and voltage-clamp recordings made from the same neuron in a living mouse. Current-clamp mode can record membrane potential changes and voltage-clamp mode can dissect synaptic currents into excitatory and inhibitory components by holding the cell membrane potential at different levels. The synaptic contributions to orientation selectivity can then be compared and quantified. The right panel depicts the orientation tuning curve for excitatory input (Ex), inhibitory input (In) and membrane potential (Vm). Error bar = SEM. The tuning width (delta) is denoted in the inset. *p < 0.01, paired t-test. Modified from Liu et al. (2011), with permission.
Mentions: Compared with the blind-patch procedure, cell location and morphology can be visualized during the recording session in visually guided patch-clamp, which is very useful for recording from specific neurons (e.g., sparsely distributed inhibitory interneurons). To visualize a target neuron in vivo, the cell needs to be either brightened or shadowed. The brightening method uses transgenic or viral methods to visualize the neurons by adding fluorescent protein to the cell membrane (Trachtenberg et al., 2002; Komai et al., 2006; Häusser and Margrie, 2014; Li et al., 2014a). Then, a pipette filled with fluorescent dye can be used guide the hunt for the target neuron (Figure 2A). In the shadowing method, the extracellular matrix surrounding the target region is perfused with a fluorescent dye and brightened; thus, target neurons can be visualized by negative signals (Kitamura et al., 2008).

Bottom Line: With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals.This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions.The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Chongqing Key Laboratory of Neurobiology, Third Military Medical University Chongqing, China.

ABSTRACT
Neuronal activity is dominated by synaptic inputs from excitatory or inhibitory neural circuits. With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals. This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions. Combining in vivo patch-clamp recording with other techniques, such as two-photon imaging or optogenetics, can provide even clearer functional dissection of the synaptic contributions of different neurons or nuclei. Here, we summarized current applications and recent research progress using the in vivo patch-clamp recording method and focused on its role in the functional dissection of different synaptic inputs. The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed.

No MeSH data available.


Related in: MedlinePlus