Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia

Tomina, Yusuke; Wagenaar, Daniel A.

doi:10.21769/bioprotoc.2751

Cited by 9 publications

(7 citation statements)

References 7 publications

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“…This network is reminiscent of a fast-conducting system (Mistick, 1978), but its causal role in any behavior is unclear (Sahley et al, 1994). To determine more neurons and their interactions involved in this preparatory network, VSD imaging with a double-sided-microscope would be an appropriate method (Tomina and Wagenaar, 2018). It would allow us to directly analyze functional relationships between neurons located on opposite surfaces and detect both action potentials and sub-threshold excitatory and inhibitory synaptic potentials (Miller et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Non-synaptic Plasticity in Leech Touch Cells

2019

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The role of Na+/K+-pumps in activity-dependent synaptic plasticity has been described in both vertebrates and invertebrates. Here, we provide evidence that the Na+/K+-pump is also involved in activity-dependent non-synaptic cellular plasticity in leech sensory neurons. We show that the resting membrane potential (RMP) of T cells hyperpolarizes in response to repeated somatic current injection, while at the same time their spike count (SC) and the input resistance (IR) increase. Our Hodgkin–Huxley-type neuron model, adjusted to physiological T cell properties, suggests that repetitive action potential discharges lead to increased Na+/K+-pump activity, which then hyperpolarizes the RMP. In consequence, a slow, non-inactivating current decreases, which is presumably mediated by voltage-dependent, low-threshold potassium channels. Closing of these putative M-type channels due to hyperpolarization of the resting potential increases the IR of the cell, leading to a larger number of spikes. By this mechanism, the response behavior switches from rapidly to slowly adapting spiking. These changes in spiking behavior also effect other T cells on the same side of the ganglion, which are connected via a combination of electrical and chemical synapses. An increased SC in the presynaptic T cell results in larger postsynaptic responses (PRs) in the other T cells. However, when the number of elicited presynaptic spikes is kept constant, the PR does not change. These results suggest that T cells change their responses in an activity-dependent manner through non-synaptic rather than synaptic plasticity. These changes might act as a gain-control mechanism. Depending on the previous activity, this gain could scale the relative impacts of synaptic inputs from other mechanoreceptors, versus the spike responses to tactile skin stimulation. This multi-tasking ability, and its flexible adaptation to previous activity, might make the T cell a key player in a preparatory network, enabling the leech to perform fast behavioral reactions to skin stimulation.

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Section: Discussionmentioning

confidence: 99%

Non-synaptic Plasticity in Leech Touch Cells

2019

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“…Detailed procedures have been described before ( Tomina and Wagenaar, 2018 ). Briefly, leeches ( Hirudo verbana , obtained from Niagara Leeches, Niagara Falls, NY) were maintained on a 12 hr:12 hr light:dark cycle in temperature-controlled aquariums filled with artificial pond water.…”

Section: Methodsmentioning

confidence: 99%

Anatomy and activity patterns in a multifunctional motor neuron and its surrounding circuits

Ashaber

Tomina

Kassraian

et al. 2021

eLife

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Dorsal Excitor motor neuron DE-3 in the medicinal leech plays three very different dynamical roles in three different behaviors. Without rewiring its anatomical connectivity, how can a motor neuron dynamically switch roles to play appropriate roles in various behaviors? We previously used voltage-sensitive dye imaging to record from DE-3 and most other neurons in the leech segmental ganglion during (fictive) swimming, crawling, and local-bend escape (Tomina and Wagenaar, 2017). Here, we repeated that experiment, then re-imaged the same ganglion using serial blockface electron microscopy and traced DE-3's processes. Further, we traced back the processes of DE-3's presynaptic partners to their respective somata. This allowed us to analyze the relationship between circuit anatomy and the activity patterns it sustains. We found that input synapses important for all of the behaviors were widely distributed over DE-3's branches, yet that functional clusters were different during (fictive) swimming vs. crawling.

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“…Procedures of dissection and dye loading for VSD imaging are described in more detail at Bio-protocol ( Tomina and Wagenaar, 2018 ). Medicinal leeches ( Hirudo verbana ) were obtained from Niagara Leeches (Niagara Falls, NY) and maintained in artificial pond water at 15°C.…”

Section: Methodsmentioning

confidence: 99%

A double-sided microscope to realize whole-ganglion imaging of membrane potential in the medicinal leech

Tomina

Wagenaar

2017

eLife

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Studies of neuronal network emergence during sensory processing and motor control are greatly facilitated by technologies that allow us to simultaneously record the membrane potential dynamics of a large population of neurons in single cell resolution. To achieve whole-brain recording with the ability to detect both small synaptic potentials and action potentials, we developed a voltage-sensitive dye (VSD) imaging technique based on a double-sided microscope that can image two sides of a nervous system simultaneously. We applied this system to the segmental ganglia of the medicinal leech. Double-sided VSD imaging enabled simultaneous recording of membrane potential events from almost all of the identifiable neurons. Using data obtained from double-sided VSD imaging, we analyzed neuronal dynamics in both sensory processing and generation of behavior and constructed functional maps for identification of neurons contributing to these processes.

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Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia

Cited by 9 publications

References 7 publications

Non-synaptic Plasticity in Leech Touch Cells

Non-synaptic Plasticity in Leech Touch Cells

Anatomy and activity patterns in a multifunctional motor neuron and its surrounding circuits

A double-sided microscope to realize whole-ganglion imaging of membrane potential in the medicinal leech

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