Method for preparing sample of nitric oxide synthase (NOS) neuron in myenteric nervous plexus of domestic pigeon

Abstract

The invention relates to a method for preparing a sample of a nitric oxide synthase (NOS) neuron in myenteric nervous plexus of a domestic pigeon. The method comprises the following steps: (1) preparing an intestinal tract to be observed; (2) taking the intestinal tract to be observed, and preparing a fixed sample; (3) cutting the fixed sample into pieces, peeling off the muscular layer, then stripping longitudinal muscle on the nerve plexus, putting into phosphate buffer for preserving, and preparing a sample to be stained; (4) washing the sample to be stained with the phosphate buffer, then dipping into confining liquid, and sealing at the room temperature; after that, dipping into a primary antibody solution, and carrying out incubation at the room temperature to prepare a primary antibody sample; (5) dipping the primary antibody sample into a goat anti-rabbit IgG solution, carrying out incubation on a shaking table, and carrying out color development under the condition of keeping out of the sun; then, dehydrating and sealing on a sheet so as to obtain the sample. The method adopts an intestinal wall full-layered sheeting-NOS immunohistochemical technique for the first time to prepare the sample of the nitric oxide synthase (NOS) neuron in the myenteric nervous plexus of the pigeon; in a preparation process, the separation between the circular muscle and the longitudinal muscle is more easily realized by injecting a muscle relaxant and stripping the longitudinal muscle before preparation of the sample.

Description

technical field [0001] The invention relates to a method for preparing NOS neuron samples in the myenteric plexus of the intestinal wall of domestic pigeons, and belongs to the technical field of biotechnology. Background technique [0002] The enteric nervous system (enteric nervous system, ENS) is composed of the myenteric plexus of the intestinal wall and the submucosal plexus, and participates in the regulation of the motility and secretion of the gastrointestinal tract. Gastrointestinal peristalsis mainly originates from and relies on smooth muscle contraction, and the excitation of smooth muscle is mainly mediated by excitatory transmitters such as acetylcholine, and the inhibition is mainly mediated by inhibitory transmitters such as nitric oxide (NO). In 1990, Bult first proposed the release of NO when the nitrogenergic neurons in the gastrointestinal tract were excited. Nitric oxide synthase (nitric oxide synthase, NOS) can catalyze L-arginine to generate NO in viv...

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As a typical representative of birds, domestic pigeons have not been studied on the morphology and charact

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