Abstract

The stomatogastric ganglion (STG) is an excellent model for studying cellular and network interactions because it contains a relatively small number of cells (approximately 25 in C. borealis) which are well characterized. The cells in the STG exhibit a broad range of outputs and are responsible for the motor actions of the stomach. The stomach contains the gastric mill which breaks down food with three internal teeth, and the pylorus which filters the food before it reaches the midgut. The STG produces two rhythmic outputs to control the gastric mill and pylorus known as central pattern generators (CPGs). Each cell in the STG can participate in one or both of these rhythms. These CPGs allow for the study of neuromodulation, homeostasis, cellular and network variability, network development, and network recovery.The dissection of the stomatogastric nervous system (STNS) from the Jonah crab (Cancer borealis) is done in two parts; the gross and fine dissection. In the gross dissection the entire stomach is dissected from the crab. During the fine dissection the STNS is extracted from the stomach using a dissection microscope and micro-dissection tools (see figure 1). The STNS includes the STG, the oesophageal ganglion (OG), and the commissural ganglia (CoG) as well as the nerves that innervate the stomach muscles. Here, we show how to perform a complete dissection of the STNS in preparation for an electrophysiology experiment where the cells in the STG would be recorded from intracellularly and the peripheral nerves would be used for extracellular recordings. The proper technique for finding the desired nerves is shown as well as our technique of desheathing the ganglion to reveal the somata and neuropil.

Highlights

  • The stomatogastric ganglion (STG) is an excellent model for studying cellular and network interactions because it contains a relatively small number of cells which are well characterized

  • The STG produces two rhythmic outputs to control the gastric mill and pylorus known as central pattern generators (CPGs)

  • Each cell in the STG can participate in one or both of these rhythms. These CPGs allow for the study of neuromodulation, homeostasis, cellular and network variability, network development, and network recovery

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Summary

Gross Dissection

Bone-cutting scissors, small scissors, toothed forceps, a spatula with a tapered edge, a black Sylgard-coated dish, and insect pins are needed. Use the rongeurs to break away the parts of the carapace that have been separated from the tissue. 9. On the ventral side of the carapace, pull out the mandibles one at a time by anchoring the rongeurs onto a mandible, twisting them outwards to dislodge the mandible from the attached epistome, and pulling it away. Cut the labrum (“upper lip”) away from the epistome making sure not to leave any tissue attached. Use the small scissors to separate the ventral part of the stomach from the tissue lining the carapace. Still holding the stomach by the lip with forceps, rest the stomach against the palm of the hand and pour saline into the mouth This expands the stomach and makes it easier to cut open.

Fine Dissection
Results
Discussion
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