Rapid Killing of Single Neurons by Irradiation of Intracellularly Injected Dye

Miller, John P.; Selverston, Allen I.

doi:10.1126/science.386514

Cited by 333 publications

(137 citation statements)

References 9 publications

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“…Standard intracellular techniques were used for voltage recordings using KC l-filled (3 M, 15-25 M⍀) microelectrodes. The AB -2xPD subnetwork was isolated by photoinactivation (Miller and Selverston, 1979) of the V D neuron in addition to pharmacological blockade (10 Ϫ5 M picrotoxin) of the L P 3 PD synapse (Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

“…A main advantage of the pyloric network is the ability to study isolated subsets of neurons, through pharmacological blockade of synapses (Bidaut, 1980) and photoinactivation to eliminate individual neurons ( Fig. 1) (Miller and Selverston, 1979;Selverston and Miller, 1980). Practically all the cellular and synaptic components of the pyloric network are targets of monoamine modulation: dopamine (DA), octopamine (Oct), and serotonin (5HT) have been found to have a bewildering variety of effects on the intrinsic electrical properties of all pyloric neurons, as well as on synaptic transmission between network members (Flamm and Harris-Warrick, 1986a,b; Harris-Warrick and Flamm, 1986Flamm, , 1987Harris-Warrick et al, 1995a,b;Johnson et al, 1995;Johnson and Harris-Warrick, 1997;.…”

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confidence: 99%

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Monoamine Control of the Pacemaker Kernel and Cycle Frequency in the Lobster Pyloric Network

Ayali

Harris‐Warrick

1999

J. Neurosci.

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The monoamines dopamine (DA), serotonin (5HT), and octopamine (Oct) can each sculpt a unique motor pattern from the pyloric network in the stomatogastric ganglion (STG) of the spiny lobster Panulirus interruptus. In this paper we investigate the contribution of individual network components in determining the specific amine-induced cycle frequency. We used photoinactivation of identified neurons and pharmacological blockade of synapses to isolate the anterior burster (AB) and pyloric dilator (PD) neurons. Bath application of DA, 5HT, or Oct enhanced cycle frequency in an isolated AB neuron, with DA generating the most rapid oscillations and Oct the slowest. When an AB-PD or AB-2xPD subnetworks were tested, DA often reduced the ongoing cycle frequency, whereas 5HT and Oct both evoked similar accelerations in cycle frequency. However, in the intact pyloric network, both DA and Oct either reduced or did not alter the cycle frequency, whereas 5HT continued to enhance the cycle frequency as before. Our results show that the major target of 5HT in altering the pyloric cycle frequency is the AB neuron, whereas DA's effects on the AB-2xPD subnetwork are critical in understanding its modulation of the cycle frequency. Octopamine's effects on cycle frequency require an understanding of its modulation of the feedback inhibition to the AB-PD group from the lateral pyloric neuron, which constrains the pacemaker group to oscillate more slowly than it would alone. We have thus demonstrated that the relative importance of the different network components in determining the final cycle frequency is not fixed but can vary under different modulatory conditions.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Monoamine Control of the Pacemaker Kernel and Cycle Frequency in the Lobster Pyloric Network

Ayali

Harris‐Warrick

1999

J. Neurosci.

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“…To determine if this was the case, we back-filled neuromodulatory axon terminals overnight with rhodamine-conjugated dextran beads or Lucifer yellow and illuminated the STG the next day to photoablate the terminals (Miller and Selverston 1979). We then recorded from the preparations overnight to determine if bouts occurred and the preparation recovered.…”

Section: Photoablation Of Neuromodulatory Terminals Had No Effect On mentioning

confidence: 99%

Episodic Bouts of Activity Accompany Recovery of Rhythmic Output By a Neuromodulator- and Activity-Deprived Adult Neural Network

Luther

Robie

Yarotsky

et al. 2003

Journal of Neurophysiology

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. Episodic bouts of activity accompany recovery of rhythmic output by a neuromodulator-and activity-deprived adult neural network. J Neurophysiol 90: 2720 -2730, 2003. First published July 2, 2003 10.1152/jn.00370.2003. The pyloric rhythm of the stomatogastric ganglion of the crab, Cancer borealis, slows or stops when descending modulatory inputs are acutely removed. However, the rhythm spontaneously resumes after one or more days in the absence of neuromodulatory input. We recorded continuously for days to characterize quantitatively this recovery process. Activity bouts lasting 40 -900 s began several hours after removal of neuromodulatory input and were followed by stable rhythm recovery after 1-4 days. Bout duration was not related to the intervals (0.3-800 min) between bouts. During an individual bout, the frequency rapidly increased and then decreased more slowly. Photoablation of back-filled neuromodulatory terminals in the stomatogastric ganglion (STG) neuropil had no effect on activity bouts or recovery, suggesting that these processes are intrinsic to the STG neuronal network. After removal of neuromodulatory input, the phase relationships of the components of the triphasic pyloric rhythm were altered, and then over time the phase relationships moved toward their control values. Although at low pyloric rhythm frequency the phase relationships among pyloric network neurons depended on frequency, the changes in frequency during recovery did not completely account for the change in phase seen after rhythm recovery. We suggest that activity bouts represent underlying mechanisms controlling the restructuring of the pyloric network to allow resumption of an appropriate output after removal of neuromodulatory input.

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“…The technique used in these experiments was a modification of the one developed by Miller and Selverston (1979) for killing whole cells in the stomatogastric ganglion and is described in detail elsewhere Kater et al, 1986). The light source was a 10 mW heliumcadmium laser (Liconix) directed through the optics of a stereo dissecting microscope (Wild MSAPO) using a specially designed epifluorescence attachment.…”

Section: Photoinactivation Of Individual Dendritesmentioning

confidence: 99%

Integrative mechanisms controlling directional sensitivity of an identified sensory interneuron

Jacobs¹,

Miller²,

Murphey³

1986

J. Neurosci.

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Several identified interneurons in the cricket cereal afferent system display directional sensitivity to wind stimuli: the spike frequency of these cells depends on the wind direction with respect to the animal's body. Factors determining the directional sensitivity of one of these identified interneurons (interneuron 10-3) were studied in detail. This cell has 3 dendritic branches that arborize in 3 distinct regions of the terminal abdominal ganglion. Using 2 independent methods, it was demonstrated that the dendrites have different receptive fields to wind stimuli. First, small patches of filiform hairs, whose afferents projected to individual dendrites, were isolated and selectively stimulated. In each case the response of the cell matched the receptive field of the afferents in the patch. Second, a laser beam directed through the stereo dissecting microscope was used to photoinactivate small portions of the cell in situ during intracellular recording. By isolating or ablating individual dendrites, the contributions of each of the 3 dendrites to the overall receptive field were assessed. Although the receptive field of the whole cell could be predicted by a summation of the receptive fields of all 3 dendrites, the precise directional sensitivity of the cell could not be predicted by a simple linear summation of the receptive fields of each dendrite. Two factors were found to account for this nonlinearity of summation. The first factor was polysynaptic inhibition from other interneurons within the terminal abdominal ganglion. Wind directions that activate inhibition in interneuron 10-3 were identified, and the specific classes of filiform afferents that activate the inhibitory pathway were determined. The net effect of the inhibition was to "sharpen" the directional sensitivity of 10-3 by selectively decreasing the cell's response to specific excitatory inputs. The second factor that contributed to directional sensitivity was the complex electroanatomy of the interneuron. The probable location of the spikeinitiating zone (SIZ) was determined by using the laser photoinactivation technique. The relative efficacies of synaptic inputs onto the 3 different branches were then interpreted with respect to their different electrotonic distances from the SIZ. On the basis of the data obtained in this report, we present a qualitative model for the basis of directional sensitivity in this cell.The relationship between the structural and functional properties of neurons has long interested theoretical and experimental neurobiologists. In very few instances, however, has it been possible to test directly how the distribution of excitatory and inhibitory inputs to different dendritic regions of a cell results in an integrated output that is relevant to the behavior of the animal. These anatomical relationships are of special interest

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Rapid Killing of Single Neurons by Irradiation of Intracellularly Injected Dye

Abstract: A simple technique for rapidly killing all or part of single neurons consists of filling the cell with Lucifer Yellow CH and irradiating all or part of it with intense blue light. Such treatment kills the irradiated part of the cell within a few minutes. Adjacent cells are not affected.

Cited by 333 publications

References 9 publications

Monoamine Control of the Pacemaker Kernel and Cycle Frequency in the Lobster Pyloric Network

Monoamine Control of the Pacemaker Kernel and Cycle Frequency in the Lobster Pyloric Network

Episodic Bouts of Activity Accompany Recovery of Rhythmic Output By a Neuromodulator- and Activity-Deprived Adult Neural Network

Integrative mechanisms controlling directional sensitivity of an identified sensory interneuron

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