Name: Cancer Borealis Stomatogastric Nervous System Dissection (Video) | JoVE Journal
Uploaded: 2009-03-23T00:00:00.000+00:00
Duration: 18 min 37 s
Description: Learn to dissect the stomatogastric nervous system from Cancer borealis for electrophysiological studies. This protocol prepares the STG and associated nerves for intracellular and extracellular recordings.

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5 minutes

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.

1. Gross Dissection
<ol>
	<li>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.</li>
	<li>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.</li>
	<li>Remove the crab from the ice bucket by its hind leg to prevent it from pinching and place it in the dissection pan.</li>
	<li>Remove the claws first by twisting each one inward. Then remove each leg using the same twisting motion.</li>
	<li>Using the rongeurs, remove the maxilla covering the mouth. Also remove the soft maxillule.</li>
	<li>Using the rongeurs again, start from the lateral posterior end of the carapace and get a good grip on the edge of it. Clamp down with the rongeurs and break that piece of carapace off. Make sure to remove enough of the carapace to allow the spatula to enter the opening. Continue removing the edge of the carapace up to the eyes and repeat on the other side.</li>
	<li>With the tapered side of the spatula, gently scrape the soft inner tissue away from the roof of the dorsal carapace. Keep the spatula in contact with the carapace to avoid damaging the STNS. Use the rongeurs to break away the parts of the carapace that have been separated from the tissue. Also separate some of the soft tissue from the ventral carapace making sure to separate any adductor muscles attaching the tissue to the carapace.</li>
	<li>Break away the dorsal carapace as far posterior as the heart where there are two small ossicles protruding from the carapace. Cut as far anterior as possible. Stabilize the cephalon (the “face”) with one finger while breaking away the most anterior parts of the dorsal carapace.</li>
	<li>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. Two long ossicles with muscles should be attached to the extracted mandible if done properly.</li>
	<li>Separate away the tissue from the sides of the cephalon near the eyes using the spatula. With small dissection scissors, cut across the tissue as near to the ridge of ossicle as possible layer by layer.</li>
	<li>Rest the crab against a wall of the dissection pan and prop it up securing it in place (the claws may be used for this). Cut the labrum (“upper lip”) away from the epistome making sure not to leave any tissue attached. Using bone-cutting scissors make two diagonal cuts into the ventral carapace to remove the cephalon.</li>
	<li>Take the labrum with toothed forceps and cut the labium (“lower lip”) away from the ossicle on the ventral side. Then gently lift the lip away from the remaining ventral carapace. Use the small scissors to separate the ventral part of the stomach from the tissue lining the carapace. When the pyloric ampullae are visible, cut the stomach away from the tissue on either side of it until the stomach is extracted from the crab.</li>
	<li>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. Cut from the opening at the mouth down between the pylorus right through the ossicle between the ampullae. Then cut diagonally down through the two cardial branches. Cut off the tips of the three teeth inside of the stomach.</li>
	<li>Place the stomach onto a large, black, Sylgard-coated dish with the inside of the stomach down. Pin down the five corners of the preparation with insect pins and fill the dish with cold saline, immersing the entire preparation (see figure 1).</li>
</ol>
2. Fine Dissection
<ol>
	<li>, Micro-dissection tools are needed for the fine dissection. In particular, forceps and spring scissors are needed as well as a pin holder with a tungsten needle. A clear, Sylgard-coated, petri dish and some fine wire pins should also have been prepared.</li>
	<li>Beginning on the lowest magnification, start by pinning down the flaps near the lip that used to be the mouth opening. Rearrange the rest of the pins to make the preparation taut and pin any other areas necessary.</li>
	<li>Carefully lift the spotted hypodermis covering the lower part of the preparation and trim it across leaving a patch of hypodermis with two white dots.</li>
	<li>Peel up the yellow, wispy, membranous tissue near the bottom of the preparation. The mvns are weakly attached to the underside of this tissue. Carefully separate the mvns from the yellow tissue as it is pulled up. Follow the yellow tissue up and cut alongside the brain.</li>
	<li>Follow the thick processes exiting the brain that extend laterally. These are the commisural nerves. Once the end of the commissural nerve is reached, remove the mass of spongy, off-white tissue that has been cut away from the side of the brain.</li>
	<li>Where the ions and sons meet the CoGs, cut away the muscle and tissue to reveal the length of the ions and sons.</li>
	<li>Return to the spotted hypodermis patch and cut it off. This should leave an opening in the artery surrounding the STG. Cut through the opening to separate the two muscles that flank the STG. Lift the ends of these muscles and cut right underneath them to separate them from the artery and to expose the alns and agns. Separate the flanking muscles as far up as the cartilage lying over the stn.</li>
	<li>Cut away the arterial tissue surrounding the STG down to where the mvns, dgn, and dvn meet. Work down the dvn to the lvns and cut through the whitish, delicate tissue covering the lvns.</li>
	<li>Locate the psn over the ossicle near the bottom of the preparation. The psn can be used to locate the lvn and dlvn junction.</li>
	<li>Next follow the dlvn down to locate the pyn and pdn. The pyn often wraps over the pyloric ampulla to the opening. The pdn forks off of the dlvn and is between the pyloric ampulla and the cardio pyloric valve muscles. Leave some muscle attached to one of these nerves because they can be easily confused when the preparation is transferred to the clear Sylgard dish.</li>
	<li>Once all of the nerves have been uncovered, sever any remaining connections between the STNS and the tissue and unwanted nerves of the stomach. Carefully move the STNS away from the rest of the stomach, cutting any missed connections.</li>
	<li>Condition the clear Sylgard dish with the remaining mass of stomach tissue by rubbing it over the surface of the Sylgard until the Sylgard no longer feels too sticky or dry. Sylgard is hydrophobic and the STNS will adhere strongly to it if the Sylgard-coated dish is not conditioned in this manner.</li>
	<li>Add some cold saline to the dish. Grab the upper ends of the commissural nerves with forceps and bring the STNS to the clear Sylgard dish.</li>
	<li>Pin the STNS down onto the Sylgard. Sever the brain from the commissural nerves and use the minuten pins to secure the four ends of the CoGs down first. Ensure that the preparation is right side up by checking that the ivn is pointing up away from the Sylgard. The dgn should exit slightly beneath the STG when the STG is right-side up. Pin the rest of the nerve ends down with the fine wire pins.</li>
	<li>Clean away any remaining muscle or tissue from the STNS.</li>
	<li>Desheath the STG using a pin holder and a slightly hooked, fine, tungsten needle. Make a small hole in a corner of the STG sheath away from the cell bodies and use that hole to get between the layers of sheath. Carefully cut a flap of separated sheath to expose the STG neuropil and cell bodies.</li>
	<li>Pin any flaps of sheath down to increase accessibility to the cell bodies and to stabilize the STG for intracellular recordings. Re-pin the rest of the STNS ensuring that each nerve is taut and well spaced for extracellular recording (see figure 3).</li>
</ol>
3. Results
<ol>
	<li>Ideally, all of the nerves should be free of nicks and damage, particularly the ones that will be recorded from. None of the nerves should be tangled or twisted. The STG should be intact with all of the cells arranged in a beard formation around the neuropil. The intact STNS is bilaterally symmetrical and looks like a homunculus with the lvns as the legs, the mvns as the arms and the anterior end as the head.</li>
</ol>
Abbreviations:
<table border="1">
	<tbody>
		<tr>
			<td>STNS</td>
			<td>Stomatogastric nervous system</td>
		</tr>
		<tr>
			<td>STG</td>
			<td>Stomatogastric ganglion</td>
		</tr>
		<tr>
			<td>CoG</td>
			<td>Commissural ganglion</td>
		</tr>
		<tr>
			<td>OG</td>
			<td>Oesophageal ganglion</td>
		</tr>
		<tr>
			<td>mvn</td>
			<td>Median ventricular nerve</td>
		</tr>
		<tr>
			<td>ion</td>
			<td>Inferior oesophageal nerve</td>
		</tr>
		<tr>
			<td>son</td>
			<td>Superior oesophageal nerve</td>
		</tr>
		<tr>
			<td>agn</td>
			<td>Anterior gastric nerve</td>
		</tr>
		<tr>
			<td>aln</td>
			<td>Anterior lateral nerve</td>
		</tr>
		<tr>
			<td>dgn</td>
			<td>Dorsal gastric nerve</td>
		</tr>
		<tr>
			<td>dvn</td>
			<td>Dorsal ventricular nerve</td>
		</tr>
		<tr>
			<td>lvn</td>
			<td>Lateral ventricular nerve</td>
		</tr>
		<tr>
			<td>psn</td>
			<td>Posterior stomach nerve</td>
		</tr>
		<tr>
			<td>pyn</td>
			<td>Pyloric nerve</td>
		</tr>
		<tr>
			<td>pdn</td>
			<td>Pyloric dilator nerve</td>
		</tr>
		<tr>
			<td>dlvn</td>
			<td>Dorsal branch of the lateral ventricular nerve</td>
		</tr>
		<tr>
			<td>ivn</td>
			<td>Inferior ventricular nerve</td>
		</tr>
	</tbody>
</table>
<img alt="Dissection diagram of crustacean nervous system, showing nerve pathways and connections." src="/files/ftp_upload/1207/1207fig1.jpg" data-altupdatedat="1787630408000" data-alt="Figure 1">
Figure 1: Diagram illustrating the approximate placement of the STNS in the stomach prior to dissection.
<img alt="Crustacean gastric and pyloric muscle diagram; muscle groups labeled with anatomical abbreviations." src="/files/ftp_upload/1207/1207fig2.jpg" data-altupdatedat="1787630408000" data-alt="Figure 2">
Figure 2: Diagram of the various muscles in the lower part of the stomach and an overlay of the STNS in light green. Each cell type in the STG is listed with the muscles they are known to innervate. This map is helpful when planning to dissect a nerve which carries a signal from a given cell (courtesy of the Nadim lab).
<img alt="Neuronal recording diagram; intracellular soma, extracellular nerve recordings in STG setup." src="/files/ftp_upload/old/1207_2009_1_12_5/figure_3.jpg" data-altupdatedat="1787630408000" data-alt="Figure 3"> 
Figure 3: Schematic diagram of the STNS with intracellular electrodes and extracellular wells (from Marder and Bucher, 2007).

<ol><li>Harris-Warrick, R. M., Marder, E., Selverston, A. I., Moulins, M. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aHarris-Warrick%2C+RM+%22Dynamic+Biological+Networks%3A+The+stomatogastric+nervous+system%22">Dynamic Biological Networks: The stomatogastric nervous system</a>. , MIT Press. Boston. (1992).</li>
<li>Marder, E., Bucher, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Understanding+circuit+dynamics+using+the+stomatogastric+nervous+system+of+lobsters+and+crabs%5BTitle%5D)+AND+%22Annu.+Rev.+Physiol%22%5BJournal%5D)">Understanding circuit dynamics using the stomatogastric nervous system of lobsters and crabs.</a> Annu. Rev. Physiol. 69, 291-316 (2007).</li>
<li>Maynard, D. M., Dando, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+structure+of+the+stomatogastric+neuromuscular+system+in+Callinectes+sapidus%2C+Homarus+Americanus+and+Panulirus+argus+%28Decapoda+crustacean%29%5BTitle%5D)+AND+%22Philos.+Trans.+R.+Soc.+Lond.+B+Biol.+Sci%22%5BJournal%5D)">The structure of the stomatogastric neuromuscular system in Callinectes sapidus, Homarus Americanus and Panulirus argus (Decapoda crustacean).</a> Philos. Trans. R. Soc. Lond. B Biol. Sci. 268 (892), 161-220 (1974).</li>
<li>Selverston, A. I., Russell, D. F., Miller, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+Stomatogastric+Nervous+System%3A+Structure+and+function+of+a+small+neural+network%5BTitle%5D)+AND+%22Prog.+Neurobiology%22%5BJournal%5D)">The Stomatogastric Nervous System: Structure and function of a small neural network.</a> Prog. Neurobiology. 7, 215-290 (1976).</li>
</ol>

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...

<table><tbody><tr><td>Black Sylgard 170</td><td>Other</td><td>Dow Corning</td><td>170</td><td>Gross Dissection</td></tr><tr><td>Fine Scissors</td><td>Surgery</td><td>Fine Science Tools</td><td>14090-09</td><td></td></tr><tr><td>Insect Pins size 6</td><td>Surgery</td><td>Fine Science Tools</td><td>26000-65</td><td></td></tr><tr><td>Mayo Scissors</td><td>Surgery</td><td>Fine Science Tools</td><td>14110-17</td><td></td></tr><tr><td>Micro-Spatula</td><td>Surgery</td><td>WARD&amp;rsquo;s Natural Science</td><td>15 V 4313</td><td></td></tr><tr><td>Rongeurs</td><td>Surgery</td><td>Fine Science Tools</td><td>16000-14</td><td></td></tr><tr><td>Tissue Forceps</td><td>Surgery</td><td>Fine Science Tools</td><td>11021-12</td><td></td></tr><tr><td>Fine Tungsten Wire (0.001 in. diameter)</td><td>Surgery</td><td>California Fine Wire Company</td><td></td><td>Fine Dissection</td></tr><tr><td>Forceps #5</td><td>Surgery</td><td>Fine Science Tools</td><td>91150-20</td><td></td></tr><tr><td>Minuten Pins</td><td>Surgery</td><td>Fine Science Tools</td><td>26002-10</td><td></td></tr><tr><td>Moria Spring Scissors (7mm blades)</td><td>Surgery</td><td>Fine Science Tools</td><td>15370-52</td><td></td></tr><tr><td>Petri Dish (100x15mm)</td><td>Other</td><td>Fisher Scientific</td><td>08-757-12</td><td></td></tr><tr><td>Pin Holder</td><td>Surgery</td><td>Fine Science Tools</td><td>26018-17</td><td></td></tr><tr><td>Super fine forceps #5SF</td><td>Surgery</td><td>Fine Science Tools</td><td>11252-00</td><td></td></tr><tr><td>Sylgard 182</td><td>Other</td><td>Dow Corning</td><td>182</td><td></td></tr><tr><td>Tungsten Needles</td><td>Surgery</td><td>Fine Science Tools</td><td>10130-05</td><td></td></tr><tr><td>Dissecting Microscope</td><td>Microscope</td><td>Wild</td><td>MC3</td><td>Misc.</td></tr><tr><td>Jonah Crab (&lt;em&gt;C. borealis&lt;/em&gt;)</td><td>Animal</td><td>Commercial Lobster</td><td></td><td></td></tr></tbody></table>

cancer borealis stomatogastric nervous system dissection

Cancer Borealis Stomatogastric Nervous System Dissection

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.

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.

The stomatogastric ganglion (STG) is an excellent model for studying cellular and network interactions because it contains a relatively small number of ...

This procedure begins with a gross dissection of the crab stomach in which the appendages and dorsal carpus are removed from the crab. The entire stomach is dissected from the animal. The stomach is cut open and pinned flat to a black sard dish.In preparation for the fine dissection, the Steamatic gastric nervous system is dissected from the stomach and is transferred to a clear Sard coated Petri dish where it is des sheathed in preparation for an electrophysiology experiment. Hi, my name is Rachel Au from the Martyr Lab at Brandeis University. And I'm Gabrielle Gutierrez, also from the Martyr Lab.Today we will show you a procedure for dissecting the somatic gastric nervous system from the Crab Cancer Borealis. We use the crab St NS in our laboratory for our electrophysiology, immunohistochemistry, and cell culture experiments. Using this neural network, we can investigate several fundamental principles of neurobiology, such as neuromodulation homeostasis, network variability, network development, and network recovery.So let's get started. Before starting the dissection, the crab must be numbed by burying it nice for 30 minutes. Be sure to surround it on all sides with ice.The ice steals the crab's, sensory and motor systems, but the crab might still be responsive when removed, while the crab chills. Prepare for the gross dissection by assembling the equipment required for this part of the protocol. The dissection pan Ron jaws bone cutting scissors, small scissors tooth forceps, a spatula with a tapered edge, a black sill guard coated dish, and insect pins Before beginning.Prepare yourself by dawning gloves. Once 30 minutes have elapsed, remove the crab from the ice bucket by its hin leg. To avoid being pinched and place in the dissection pan first, remove the claws by twisting each one inward.Next, remove each leg using the same twisting motion. Now that the animal is appendage list, use the Ron Jos to remove the maxilla covering the mouth and the soft maxil Using the Ron Jos again, start from the lateral posterior end of the carus and get a good grip on the edge of it. Clamp down with the Ron Jos and break that piece of carpus off.Make sure that you remove enough of the carpus to allow the spatula to enter the opening. Continue removing the edge of the carpus up to the eyes and repeat on the other side. With the tapered side of the spatula, gently scrape the soft inner tissue away from the roof of the dorsal carpus.While you do this, keep the spatula in contact with the carus. To avoid damaging the STNS, use the Ron Jos to break away parts of the carus that have been separated from the tissue. Break away the dorsal carus up to where there are two small S protruding from the carus.Cut as far anteriorly as possible. Stabilize cephalon with one finger while breaking away the most anterior parts of the dorsal carus. Separate some of the soft tissue from the ventral carus.Make sure to separate away any adductor muscles attaching the tissue to the carus on the ventral side of the carus. Pull out the mandibles one at a time by anchoring the wrong jaws onto a mandible twisting outwards and pulling away two long obstacles with muscles should be attached to the extracted mandible. If done properly, separate away the tissue from the sides of the cephalon near the eyes.Using the spatula with small dissection scissors, layer by layer cut across the tissue right next to the ridge of ossicle. Prop up what remains of the crab against the dissection pan. Using the detached claws, detach all of the labrum from the epistle.Using the small scissors using bone cutting scissors, make two diagonal cuts into the ventral carpus. To remove the cephalon, now take the labrum with tooth forceps and cut the labium away from the osci on the ventral side. Gently lift the lip away from the remaining ventral carus.Use small scissors to separate the ventral part of the stomach from the tissue lining the carus. When the pori canula are visible, cut the stomach away from the tissue on either side of it until the stomach is extracted from the crab, holding the stomach by the lip with forceps. Rest the stomach against your palm and pour saline into the mouth.This will expand the stomach and make it easier to cut open using the small scissors. Cut from the opening of the mouth down between the ula right through the obstacle. Then cut diagonally down through the two cardio branches.Lastly, cut off the tips of the three teeth inside the stomach. Place the stomach onto a black sard coated dish with the inside of the stomach down to prepare for the fine dissection. Pinned down the five corners of the stomach with insect pins and immerse the preparation in cold saline.The fine dissection is done under the stereoscope and requires forceps and spring scissors, as well as a pin holder with a tungsten needle. A clear sill guard coated Petri dish and some fine wire pins should already be prepared beginning on the lowest magnification, pin down the flaps near the lip that used to be the mouth opening. Now make the preparation taut by rearranging and adding pins as needed.With forceps, carefully lift the spotted hypodermis covering the lower part of the preparation and trim it across What should remain as a patch of hypodermis with two white dots. Now peel up the yellow wispy membranous tissue near the bottom of the preparation. The M vns are weak attached to the underside of this tissue.Using the blade of the scissors carefully separate the M VNS from the yellow tissue as it is pulled up. Cut away the yellow tissue alongside the few visible thick white processes of the brain. Most of the brain is currently buried in tissue.Follow the processes that extend laterally and toward the lip. These are the two commissural nerves. Once the end of the commissural nerve is found, remove the massive spongy off white tissue that has been cut away from the side of the brain.Locate the iOS and sos at the junction where they meet the commis ganglia. Cut away the muscle and tissue to reveal the length of the IO ns and so ns, these two nerves run medially from the commis ganglia and meet the esophageal nerve. Return to the spotted hypodermis patch and cut it off.This should leave an opening in the artery surrounding the STG Cut through the opening to separate the two muscles that flank the STG. Lift the ends of these muscles and cut underneath them to separate them from the artery. This step exposes the ALNs.Remove the flanking muscles as far up as the cartilage line over the STN. The TN is a nerve running up from the STG and it has a slight bulge. Next, cut away the arterial tissue surrounding the STG down to where the M vns DGN and DVN meet.Follow the DVN as it splits into the two LVNs. Cut through the whiteish delicate tissue until each LVN is completely exposed and meets the DLVN. Locate the PSN over the Osco.Near the bottom of the preparation, the PSN can be used to locate the LVN and DLVN junction. So the DV DLV goes this way. It comes down here.Yep.Then it splits and goes this way.Right. And so what I, what I normally do is I find them down here and I sort of work my way back up The PDN forks off of the DLVN and is between the ULA and the cardio Pylori valve muscles.Yeah. So here are the ampule ula, ula ampule, the P, the PYN right here.Be sure to leave some muscle attached to one of these nerves as a marker to identify them from each other. Once all of the nerves have been uncovered, sever any remaining connections between the STNS and the stomach. Finally, carefully move the STNS away from the rest of the stomach.Cutting any missed connections. Prepare the clear S guard dish by rubbing some stomach tissue over it until it feels wet and less sticky. The STNS will adhere strongly to the sogar.If the dish is not conditioned in this manner, then add some fresh cold saline to the dish. Grab the upper ends of the commissional nerves with forceps and move the st NS to the clear sogar dish. The brain will still be attached to the commissional nerves to prevent untangling.Now, prepare the STNS for pinning by severing the brain from the Commis nerves. Use minuchin pins to secure the four ends of the Commis nerves to the sill guard. Ensure that the preparation is right side up by checking that the IVN is pointing up and that the DGN exits slightly beneath the STG pin.The rest of the nerve ends down with the fine wire pins Using fine forceps and scissors. Carefully clean away any remaining muscle or tissue from the STNS. The STG must now be de sheathed using a pin holder in a slightly hooked fine tungsten needle.Make a small hole in the corner of the STG sheath away from the cell bodies. Gently poke the tungsten needle into the hole and get it between the sheath and the Samara. Cut or gently tear a flap of separated sheath to expose the STG neuro pill and cell bodies.Pin any flaps of sheath down to increase accessibility to the cell bodies and to stabilize the STG for intracellular recordings. Rein the rest of the STNS ensuring that each nerve is taught and well spaced for extracellular recording. Ideally, all of the nerves should be free of necks and damage, particularly the ones you will record from.None of the nerves should be tangled or twisted. The STG should be intact with all of its cells arranged in a beard formation around the NI bill. The TNS was bilaterally symmetrical and looks like a homunculus with the LVNs as the legs.The M vns is the arms and the anterior end is the head. We've just shown you how to dissect the somatic gastric nervous system of the crab in preparation for an electrophysiology experiment. When doing this procedure, it's important to remember to dissect the nerves you plan to record from by following the desired nerves into the muscles they're known to innervate.Ensure that the STNS is pinned taut onto the clear cigar dish with the de sheathed side of the STG facing up. So that's it. Thanks for watching and good luck with your experiments.

cancer-borealis-stomatogastric-nervous-system-dissection

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In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

Perineural invasion (PNI) is a common feature of head and neck squamous cell carcinoma (HNSCC), conferring lower survival rates. Its mechanisms are poorly understood. Utilizing neurites generated from murine dorsal root ganglia confined to a semisolid matrix, the pathways involved in the PNI of HNSCC cell lines can be investigated.

A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma

Here we present a protocol for minimally invasive surgical lesioning of muscles intrinsic to the feeding apparatus of the marine mollusk Aplysia californica to understand the roles of these muscles during feeding behavior.

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

This protocol presents a method for imaging neuronal population activity with single-cell resolution in non-transgenic invertebrate species using absorbance voltage-sensitive dyes and a photodiode array. This approach enables a rapid workflow, wherein imaging and analysis can be pursued over the course of a single day.

Photodiode-Based Optical Imaging for Recording Network Dynamics with Single-Neuron Resolution in Non-Transgenic Invertebrates

The present protocol describes a mouse model of the ablation of adrenergic innervation by identifying and resecting the superior cervical ganglion.

Surgical Technique for Superior Cervical Ganglionectomy in a Murine Model

Learn to dissect the stomatogastric nervous system from Cancer borealis for electrophysiological studies. This protocol prepares the STG and associated nerves for intracellular and extracellular recordings.

Cancer Borealis Stomatogastric Nervous System Dissection

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