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Method Article

Cancer Borealis Stomatogastric Nervous System Dissection

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DOI:

10.3791/1207

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March 23rd, 2009

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In This Article

Summary

The stomatogastric nervous system (STNS) of the Jonah crab (C. borealis) can be used for electrophysiology, immunohistochemistry, and cell culture studies. The STNS extraction is done in two parts: the gross and fine dissection.

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.

Protocol

1. Gross Dissection

  1. Place a crab on ice for 30 minutes to dull the sensory and motor systems of the animal. Surround it on all sides with ice. Gloves should be worn when handling the crab.
  2. Prepare for the gross dissection by laying out the dissection pan and the tools you will use. Rongeurs, bone-cutting scissors, small scissors, toothed forceps, a spatula with a tapered edge, a black Sylgard-coated dish, and insect pins are needed.
  3. Remove the crab from the ice bucket by its hind leg to prevent it from pinching and place it in the dissection pan.
  4. Remove the claws first by twisting each one inward. Then remove each leg using th....

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Discussion

The STNS dissection is the first step in performing an electrophysiology or immunohistochemistry experiment, or for obtaining cells for cell culture. Regardless of the intended experiment, the gross dissection will remain unchanged. However, the fine dissection may vary. It is important to plan ahead of time which cells and which nerves you will record from. Most neurons in the STG have processes that innervate at least one muscle in the stomach (see figure 2) and are classified according to their innervated target. The.......

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Acknowledgements

We gratefully acknowledge the members of the Marder Lab. We thank our advisor, Dr. Eve Marder, for her support and encouragement. This research was supported by the National Institute of Neurological Disorders and Stroke Grants NS-17813 and NS-058110.

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Black Sylgard 170OtherDow Corning170Gross Dissection
Fine ScissorsSurgeryFine Science Tools14090-09
Insect Pins size 6SurgeryFine Science Tools26000-65
Mayo ScissorsSurgeryFine Science Tools14110-17
Micro-SpatulaSurgeryWARD’s Natural Science15 V 4313
RongeursSurgeryFine Science Tools16000-14
Tissue ForcepsSurgeryFine Science Tools11021-12
Fine Tungsten Wire (0.001 in. diameter)SurgeryCalifornia Fine Wire CompanyFine Dissection
Forceps #5SurgeryFine Science Tools91150-20
Minuten PinsSurgeryFine Science Tools26002-10
Moria Spring Scissors (7mm blades)SurgeryFine Science Tools15370-52
Petri Dish (100x15mm)OtherFisher Scientific08-757-12
Pin HolderSurgeryFine Science Tools26018-17
Super fine forceps #5SFSurgeryFine Science Tools11252-00
Sylgard 182OtherDow Corning182
Tungsten NeedlesSurgeryFine Science Tools10130-05
Dissecting MicroscopeMicroscopeWildMC3Misc.
Jonah Crab (C. borealis)AnimalCommercial Lobster

References

  1. Harris-Warrick, R. M., Marder, E., Selverston, A. I., Moulins, M. Dynamic Biological Networks: The stomatogastric nervous system. , MIT Press. Boston. (1992).
  2. Marder, E., Bucher, D. Understanding circuit dynamics using the stomatogastric nervous system of lobsters and crabs.<....

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