<em> In Vitro</em> Подготовка к Выявление и записи программ кормления двигателя с Физиологические движения в<em> Aplysia саЩогтса</em
Summary
Мы описываем технику внеклеточно записывать и стимулировать от нервов, мышц и отдельных нейронов определены<em> В пробирке</em> В то время как выявление и наблюдение за различными типами кормления поведения в кормлении аппарата<em> Aplysia</em>.
Abstract
Multifunctionality, the ability of one peripheral structure to generate multiple, distinct behaviors1, allows animals to rapidly adapt their behaviors to changing environments. The marine mollusk Aplysia californica provides a tractable system for the study of multifunctionality. During feeding, Aplysia generates several distinct types of behaviors using the same feeding apparatus, the buccal mass. The ganglia that control these behaviors contain a number of large, identified neurons that are accessible to electrophysiological study. The activity of these neurons has been described in motor programs that can be divided into two types, ingestive and egestive programs, based on the timing of neural activity that closes the food grasper relative to the neural activity that protracts or retracts the grasper2. However, in isolated ganglia, the muscle movements that would produce these behaviors are absent, making it harder to be certain whether the motor programs observed are correlates of real behaviors. In vivo, nerve and muscle recordings have been obtained corresponding to feeding programs2,3,4, but it is very difficult to directly record from individual neurons5. Additionally, in vivo, ingestive programs can be further divided into bites and swallows1,2, a distinction that is difficult to make in most previously described in vitro preparations.
The suspended buccal mass preparation (Figure 1) bridges the gap between isolated ganglia and intact animals. In this preparation, ingestive behaviors – including both biting and swallowing – and egestive behaviors (rejection) can be elicited, at the same time as individual neurons can be recorded from and stimulated using extracellular electrodes6. The feeding movements associated with these different behaviors can be recorded, quantified, and related directly to the motor programs. The motor programs in the suspended buccal mass preparation appear to be more similar to those observed in vivo than are motor programs elicited in isolated ganglia. Thus, the motor programs in this preparation can be more directly related to in vivo behavior; at the same time, individual neurons are more accessible to recording and stimulation than in intact animals. Additionally, as an intermediate step between isolated ganglia and intact animals, findings from the suspended buccal mass can aid in interpretation of data obtained in both more reduced and more intact settings. The suspended buccal mass preparation is a useful tool for characterizing the neural control of multifunctionality in Aplysia.
Protocol
Representative Results
Discussion
Предыдущая работа характеризуется Aplysia двигателя программ в интактных животных и в условиях ограниченной препараты, такие как изолированные ганглиев. В интактных животных, хотя записи отдельных нейронов было получено 5, такие эксперименты очень трудно, и электроды не могут …
Disclosures
The authors have nothing to disclose.
Acknowledgements
Это исследование было поддержано грантом NS047073 NIH и NSF гранта DMS1010434.
Materials
| Name | Company | Catalog Number | Comments |
| Sodium chloride | Fisher Scientific | S671 | Biological, Certified |
| Potassium chloride | Fisher Scientific | P217 | Certified ACS |
| Magnesium chloride hexahydrate | Acros Organics | 19753 | 99% |
| Magnesium sulfate heptahydrate | Fisher Scientific | M63 | Certified ACS |
| Calcium chloride dihydrate | Fisher Scientifc | C79 | Certified ACS |
| Glucose (dextrose) | Sigma-Aldrich | G7528 | BioXtra |
| MOPS buffer | Acros Organics | 17263 | 99% |
| Carbachol | Acros Organics | 10824 | 99% |
| Sodium hydroxide | Fisher Scientific | SS255 | Certified |
| Hydrochloric acid | Fisher Scientific | SA49 | Certified |
| Single-barreled capillary glass | A-M Systems | 6150 | |
| Flaming-Brown micropipette puller model P-80/PC | Sutter Instruments | Filament used: FT345B | |
| Enamel coated stainless steel wire | California Fine Wire | 0.001D, coating h | |
| Household Silicone II Glue | GE | ||
| Duro Quick-Gel superglue | Henkel corp. | ||
| A-M Systems model 1700 amplifier | A-M Systems | Filter settings: 300-500 Hz nerves,10-500 Hz I2 muscle | |
| Pulsemaster Multi-Channel Stimulator | World Precision Instruments | A300 | |
| Stimulus Isolator | World Precision Instruments | A360 | |
| AxoGraph X | AxoGraph Scientific | ||
| Veeder-Root Totalizing Counter | Danaher | C342-0562 | |
| Gold Connector Pins | Bulgin | SA3148/1 | |
| Gold Connector Sockets | Bulgin | SA3149/1 | |
| Sylgard 184 Silicone Elastomer | Dow Corning | ||
| 100 x 50 mm Crystalizing Dish | Pyrex | ||
| High Vacuum Grease | Dow Corning | ||
| Pipet Tips | Fisher Scientific | 21-375D | |
| Minutien Pins | Fine Science Tools | 26002-10 | |
| Modeling Clay | Sargent Art | 22-4400 | |
| Silk Sutures | Ethicon | K89OH | |
| Whisper Air Pump | Tetra | 77849 | |
| Aquarium Tubing | Eheim | 7783 | 12/16 mm |
| Elite Airstone | Hagen | A962 | |
| Vannas Spring Scissors | Fine Science Tools | 15000-08 | |
| Dumont #5 Fine Forceps | Fine Science Tools | 11254-20 | |
| Yaki Sushi Nori Seaweed | Rhee Bros | ||
| Kimwipes | Kimberly-Clark | 34155 |
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