Radzimirska Malgorzata, Kuchinka Jacek, Nowak Elzbieta, Trybus Wojciech, Szczurkowski Aleksander
Department of Medical Biology, Institute of Biology, Jan Kochanowski University in Kielce, Poland, Poland.
Folia Histochem Cytobiol. 2020;58(1):54-60. doi: 10.5603/FHC.a2020.0005. Epub 2020 Mar 23.
Cholinergic and adrenergic innervation of the pancreas in chinchilla (Chinchilla Laniger Molina) was examined in this study. The pancreas is both an exocrine and endocrine gland with autonomic and sensory innervation presented by the numerous nerve fibers and small agglomerations of nerve cells.
Investigations were performed on 16 adult chinchillas of both sexes. The material was collected immediately after death of the animals. Histochemical methods: AChE and SPG were used, in addition to routine technique of single and double immunohistochemical (IHC) staining using whole mount specimens and freezing sections with a thickness of 8 to 12 μm. In the immunofluorescence staining, primary antibodies directed against markers used to identify cholinergic - ChAT and VAChT, and adrenergic - DbH and TH neurons. Secondary antibodies were coupled to Alexa Fluor 488 and Alexa Fluor 555 fluorophores.
Histochemical studies (AChE) revealed that chinchilla pancreatic cholinergic innervation consisted of ganglionic neurocytes and numerous nerve fibers. These structures are located in the parenchyma of the exocrine part of the organ in close proximity to blood vessels and are present within the walls of the pancreatic ducts and interstitial connective tissue. A delicate fiber network around the Langerhans islets was also observed. The most numerous cholinergic structures were found in the head and tail, and the least numbers were found in the body of the pancreas. The SPG method revealed that adrenergic fibers form a network in the adventitia of blood vessels, and individual fibers run throughout the pancreatic parenchyma. Moreover, adrenergic nerve fibers were observed around the ganglionic neurocytes. This innervation was similar in all parts of the investigated organ. IHC investigations allowed observations of both the cholinergic and adrenergic activities of autonomic nerve structures. Additionally, using ChAT/DbH double staining, colocalization of these substances was observed in the fibers of the pancreatic parenchyma that passed through the cholinergic ganglia. Colocalization of VAChT and TH was found in nerve fibers of the exocrine part, in the walls of blood vessels, and in individual nerve cells. Colocalization of ChAT/DbH and VAChT/TH was observed in the single nerve cells and in the small (2-3 cell) ganglia. ChAT- and DbH-immunopositive nerve fibers were found in the area of the islets of Langerhans.
The results indicate a more intense cholinergic innervation of the chinchilla's pancreas, which is represented by both ganglia and nerve fibers, while adrenergic structures are mainly represented by fibers and only single neurocytes. This arrangement of the investigated structures in this species may imply a major role for hormonal control of exocrine secretion in rodents.
本研究对毛丝鼠(Chinchilla Laniger Molina)胰腺的胆碱能和肾上腺素能神经支配进行了检查。胰腺既是外分泌腺又是内分泌腺,有由众多神经纤维和神经细胞小聚集体构成的自主神经和感觉神经支配。
对16只成年雌雄毛丝鼠进行了研究。材料在动物死后立即采集。组织化学方法:除了使用整装标本和厚度为8至12μm的冰冻切片进行单重和双重免疫组织化学(IHC)染色的常规技术外,还使用了乙酰胆碱酯酶(AChE)和嗜银蛋白(SPG)方法。在免疫荧光染色中,使用针对用于识别胆碱能——胆碱乙酰转移酶(ChAT)和囊泡乙酰胆碱转运体(VAChT),以及肾上腺素能——多巴胺β-羟化酶(DbH)和酪氨酸羟化酶(TH)神经元的标志物的一抗。二抗与Alexa Fluor 488和Alexa Fluor 555荧光团偶联。
组织化学研究(AChE)显示,毛丝鼠胰腺胆碱能神经支配由神经节神经细胞和众多神经纤维组成。这些结构位于器官外分泌部的实质内,靠近血管,存在于胰管壁和间质结缔组织内。还观察到围绕胰岛的精细纤维网络。在胰腺头部和尾部发现的胆碱能结构最多,而在胰腺体部发现的数量最少。SPG方法显示,肾上腺素能纤维在血管外膜形成网络,个别纤维贯穿胰腺实质。此外,在神经节神经细胞周围观察到肾上腺素能神经纤维。在所研究器官的所有部位,这种神经支配都是相似的。免疫组织化学研究能够观察到自主神经结构的胆碱能和肾上腺素能活性。此外,使用ChAT/DbH双重染色,在穿过胆碱能神经节的胰腺实质纤维中观察到这些物质的共定位。在腺泡部分的神经纤维、血管壁和单个神经细胞中发现了VAChT和TH的共定位。在单个神经细胞和小(2 - 3个细胞)神经节中观察到ChAT/DbH和VAChT/TH的共定位。在胰岛区域发现了ChAT和DbH免疫阳性神经纤维。
结果表明毛丝鼠胰腺的胆碱能神经支配更为密集,由神经节和神经纤维共同代表,而肾上腺素能结构主要由纤维和仅有的单个神经细胞代表。该物种中这些被研究结构的这种排列可能意味着激素对啮齿动物外分泌分泌的控制起主要作用。