大鼠胰腺γ-氨基丁酸免疫组织化学定位

本文用ABC—GDN免疫组织化学方法,研究了γ-氨基丁酸(Gamma—Aminobutyric Acid,GABA)在大鼠胰腺的定位和分布,并用相邻切片法,观察它与胰岛素的共存关系。结果发现GABA免疫反应阳性细胞主要分布于胰腺内分泌部(胰岛)。在外分泌部亦有少许分布。大部分胰岛细胞呈GABA免疫反应阳性,集中位于胰岛的中央部。相邻连续切片免疫染色证实GABA与胰岛素共存于胰岛B细胞中。外分泌部胰腺GABA免疫反应阳性细胞,呈零散分布于腺泡和导管上皮间。本文为进一步探讨GABA在胰腺的生理作用提供了形态学依据。     

大鼠胰腺嗜铬颗粒素A分布的免疫组织化学研究

本研究用ＡＢＣ免疫组织化学方法,在Ｂｏｕｉｎ液固定的常规石蜡切片上,观察了啥铬颗粒素Ａ在大鼠胰腺内分泌细胞内的定位和分布,并用相邻切片双标记法,观察了它与胰高血糖素、胰岛素、生长抑素的共存关系。结果发现,大鼠胰腺嗜铬颗粒素Ａ样免疫反应细胞主要分布于胰岛的周边部,胰腺外分泌部的导管和腺泡等处均未见ＣｇＡ祥物质存在。用相邻薄切片免疫显色技术证明,大鼠胰腺中ＣｇＡ样物质与胰高血糖素共存。结果提示,ＣｇＡ可能是胰腺内分泌细胞的一个新的标志物,在胰腺功能调节上发挥着重要作用。     

胰岛淀粉样多肽在豚鼠胰腺分布的免疫组织化学研究

本文用免疫组织化学ＡＢＣ法,研究了胰岛淀粉样多肽（Ｉｓｌｅｔａｍｙｌｏｉｄｐｏｌｙｐｅｐｔｉｄｅ,ＩＡＰＰ或称Ａｍｙｌｉｎ）在豚鼠胰脏的分布,并用邻片免疫组织化学双标记法,观察了ＩＡＰＰ与胰岛素（Ｉｎｓｕｌｉｎ,ＩＮＳ）、生长抑素（ＳｏｍａｔｏｓｔａｔｉｎＳＳ）的共存关系。结果显示,豚鼠胰岛内绝大多数细胞都呈ＩＡＰＰ阳性免疫反应,在胰外分泌部的腺泡和导管内也散在分布有ＩＡＰＰ免疫反应阳性细胞。多数ＩＡＰＰ免疫反应阳性的细胞都显示ＩＮＳ免疫反应阳性,胰岛内少数ＩＡＰＰ阳性细胞也呈ＳＳ免疫反应阳性。说明ＩＡＰＰ主要分布在豚鼠的胰岛内．但也少量存在于外分泌部。ＩＡＰＰ主要和ＩＮＳ共存于Ｂ细胞内。但也和ＳＳ共存于Ｄ细胞内,提示ＩＡＰＰ可能通过自分泌途径调节ＩＮＳ和ＳＳ的分泌。     

人胰腺内分泌细胞中CD 57免疫反应性的表达

CD57是一种分子量为110 KD的糖蛋白,它是人类自然杀伤细胞(NK)和杀伤细胞(K)的特异性表面抗原。用它免疫小鼠产生的抗CD57单克隆抗体可特异性识别人的NK 和K 细胞表面抗原。本文应用ABC 免疫组织化学染色结合葡萄糖氧化酶DAB-硫酸镍铵显色技术研究了CD 57在人胰腺的表达和分布。结果发现,CD 57免疫反应(CD 57一IR)细胞主要分布在胰腺的内分泌部(胰岛),偶见于胰腺外分泌部的腺泡和导管上皮中。多数CD 57-IR 细胞呈典型内分泌细胞的形态特征。相邻切片法证明大多数Glu,SS,HPP 和PS 免疫反应细胞呈CD 57阳性。胰腺内神经纤维亦呈CD 57阳性。少数呈CD 57阳性的NK 和K 细胞散布在胰腺结缔组织中。本文对CD 57免疫反应性在人胰腺内分泌细胞中表达的意义进行了讨论。     

哺乳动物胰腺体部胰多肽（PP）免疫反应细胞的比较研究

采用SPA-GDN免疫组化染色技术,对人、大鼠、小鼠、豚鼠、猪、狗和猫等七种哺乳动物胰腺体部胰多肽(PP)免疫反应细胞的分布和形态进行比较研究,结果表明,上述七种动物PP细胞的分布和形态有明显的种间差异。人、大鼠和小鼠PP细胞主要位于胰岛周边部,形成环形结构,少量PP细胞散布在外分泌部的腺泡和导管;而豚鼠、猪和猫的PP细胞则主要分布在外分泌部腺泡和导管上皮间;狗的PP细胞在内、外分泌部均有分布。PP细胞的形态在上述动物间也有明显的差异,这可能与该细胞在不同动物的作用途径及功能不同有关。     

胆囊收缩素和胰岛素在发育中小鼠胰腺内的表达

目的观察昆明小鼠发育过程中胰腺内胆囊收缩素免疫反应阳性（CCK—IR）细胞和胰岛素免疫反应阳性（Ins—IR）细胞的发生、分布和形态。方法用免疫组织化学和免疫荧光双标法观察小鼠胚胎11d至出生后45d胰腺内的CCK—IR细胞和Ins—IR细胞。结果小鼠胚胎第11d,早期分化的胰腺中即出现CCK—IR细胞和Ins—IR细胞。自胚胎期至生后发育成熟,小鼠胰腺中均可见到Ins—IR细胞。CCK—IR细胞的形态不规则,与Ins—IR细胞相邻分布。在胚胎期免疫反应较强,生后胰腺内依然可见到发弱荧光的CCK—IR细胞。胰岛内还可见CCK—Ins—IR细胞。CCK—IR细胞和Ins—IR细胞主要分布于胰岛内,外分泌部的腺泡和导管处也偶可见到。结论发育中小鼠胰腺内的CCK—IR细胞具有旁分泌细胞的形态特征;免疫荧光双标法可见CCK与胰岛素共存于同一细胞内的现象。     

白细胞介素-1受体I型在正常大鼠胰腺的表达

目的研究白细胞介素-1的Ⅰ型受体(IL-1RI)蛋白在正常大鼠胰腺的表达。方法Western blotting和免疫荧光双重染色。结果胰腺组织的Western blotting结果表明,阳性反应条带出现在80 kD处,与IL-1RI分子量一致。免疫荧光双重染色证实,胰岛素(胰岛β细胞的标志物)免疫反应阳性的细胞均为IL-1RI阳性;胰腺外分泌部细胞呈胰岛素免疫反应阴性,但IL-1RI为阳性。结论大鼠胰腺胰岛的β细胞和外分泌部细胞均表达IL-1RI,提示促炎性细胞因子可能对胰腺的内、外分泌功能都有影响。     

IAPP与SS在大鼠和家兔背根神经节细胞内分布与共存的免疫组织化学研究

应用免疫组织化学ABC法,观察到在大鼠和家兔背根神经节内存在胰岛淀粉样多肽免疫阳性反应细胞.用相邻镜像邻片免疫双标记技术,证实胰岛淀粉样多肽存在于背根神经节内那些生长抑素免疫反应阳性的细胞中,说明胰岛淀粉样多肽在大鼠和家兔背根神经节内与生长抑素共存.实验结果为胰岛淀粉样多肽的胰腺外研究提供了新的资料,为进一步研究背根神经节内生物活性物质间的关系及生理作用提供了形态学证据.     

酪氨酸羟化酶在成年大鼠胰腺中的定位

目的:酪氨酸羟化酶(tyrosine hydroxylase,TH)是儿茶酚胺类递质合成的限速梅,儿茶酚胺类递质对胰腺内分泌细胞的功能具有重要的调控作用,本研究拟探讨酪氨酸羟化酶(tyrosine hydroxylase,TH)在成年大鼠整个胰腺的具体定位和表达.方法:取雄性成年大鼠胰腺,冰冻组织切片,应用免疫荧光技术观察酪氨酸羟化酶在整个胰腺中的表达分布情况,并进一步运用免疫荧光双标技术鉴定酪氨酸羟化酶是否与胰岛素、胰高血糖素、生长抑素以及胰多肽分别共定位于β细胞;α细胞;δ细胞及PP细胞,进一步确定合成酪氨酸羟化酶确切的细胞类型.结果:①在胰腺腺泡细胞胞浆中存在酪氨酸羟化酶的阳性表达颗粒.②分布于胰腺外分泌腺的神经纤维和胰岛的神经纤维中都有酪氨酸羟化酶的表达.③酪氨酸羟化酶与胰岛的四种内分泌细胞所合成的肽之间均没有共定位关系.结论:在胰腺,酪氨酸羟化酶只存在于胰腺外分泌腺的腺泡细胞胞浆内以及胰腺中的神经纤维中,而胰岛四种内分泌细胞中没有酪氨酸羟化酶,说明胰腺儿茶酚胺类神经递质一方面由胰腺外分泌部的腺泡细胞合成,另一方面来源于神经末梢的释放,而胰岛细胞不能合成儿茶酚胺类递质;该结果为进一步研究胰腺内、外分泌部之间的关系和儿茶酚胺对胰腺分泌功能的调节提供形态学证据.     

葡萄糖氧化酶—二氨基联苯胺—硫酸镍铵法在免疫酶双重染色技术中应用

作者在豚鼠胰腺组织石蜡切片的PAP免疫酶双重标记染色中,分别使用葡萄糖氧化酶-二氨基联苯胺-硫酸镍铵(Glucose oxidase-DAB-Nickel,GDN)和单纯DAB显示生长抑素(Somatostatin,SOM)及5-羟色胺(5-hpdroxytryptamin,5-HT)免疫反应细胞,获得良好的染色结果。该方法步骤简单,结果明确,背景清楚,是取得理想的免疫酶双重染色结果的新途径。     

Immunocytochemical study of gastro-entero-pancreatic (GEP) endocrine cells in the vampire bat (Desmodos rotundus)

The distribution and frequency of gastro-entero-pancreatic (GEP) endocrine cells were studied in vampire bats by immunocytochemistry. Moderate numbers of somatostatin- and a few 5-hydroxytryptamine (5-HT)- and glucagon-immunoreactive cells were seen in the fundic cecum of the stomach. Numerous gastrin- and moderate numbers of somatostatin- and 5-HT-immunoreactive cells were found in the pyloric region. Moderate numbers of 5-HT-, somatostatin-, and gastrin-immunoreactive cells also were found in BRUNNER's glands. In addition to the above-mentioned 4 immunoreactive cell types, cells immunoreactive for glicentin, secretin, cholecystokinin (CCK), gastric inhibitory peptide (GIP), and neurotensin were found in the intestine. Numerous insulin-, moderate numbers of somatostatin- and glucagon-, and a few 5-HT-immunoreactive cells were detected in the pancreatic islets with lesser numbers scattered within the exocrine pancreas. Motilin- and pancreatic polypeptide-immunoreactive cells were not observed in this study.     

Immunohistochemical study of the developing endocrine pancreas of the opossum (Didelphis virginiana)

Cells immunoreactive for insulin, glucagon, somatostatin, bovine pancreatic polypeptide and 5-hydroxytryptamine are found in the pancreas of the newborn opossum and of all later stages examined. All immunoreactive cell types are present in primary and secondary islets and within elements of the exocrine pancreas. Cells immunoreactive for glucagon, bovine pancreatic polypeptide, somatostatin and 5-hydroxytryptamine generally are confined to the periphery of secondary (intralobular) islets, whereas insulin-immunoreactive cells occupy the central region. Endocrine cells within primary (interlobular) islets are randomly scattered. A small number of pancreatic-polypeptide-immunoreactive cells are reactive for the amine 5-hydroxytryptamine also, but the reverse is not observed. The endocrine pancreas continues to differentiate and develop throughout postnatal life and into adulthood. Little difference was observed between the head and tail regions of the opossum pancreas for the measurements made.     

Impaired differentiation of endocrine and exocrine cells of the pancreas in transgenic mouse expressing the truncated type II activin receptor

Activin A is expressed in endocrine precursor cells of the fetal pancreatic anlage. To determine the physiological significance of activins in the pancreas, a transgenic mouse line expressing the truncated type II activin receptor under the control of beta-actin promoter was developed. Histological analyses of the pancreas revealed that the pancreatic islets of the transgenic mouse were small in size and were located mainly along the pancreatic ducts. Immunoreactive insulin was detected in islets, some acinar cells, and in some epithelial cells in the duct. In addition, there were abnormal endocrine cells outside the islets. The shape and the size of the endocrine cells varied and some of them were larger than islets. These cells expressed immunoreactive insulin and glucagon. In the exocrine portion, there were morphologically abnormal exocrine cells, which did not form a typical acinar structure. The cells lacked spatial polarity characteristics of acinar cells but expressed immunoreactive amylase, which was distributed diffusely in the cytoplasm. Plasma glucose concentration was normal in the transgenic mouse before and after the administration of glucose. The insulin content of the pancreas in transgenic and normal mice was nearly identical. These results suggest that activins or related ligands regulate the differentiation of the pancreatic endocrine and exocrine cells.     

中华大蟾蜍多种组织内5—羟色胺免疫染色细胞的分布

The distribution of 5-hydroxytryptamine (5-HT) immunostaining cells in the digestive tracts (hibernation and nonhibernation), the brain and other various tissues of Bufo bufo gargarizans was studied by peroxidase anti-peroxidase immunocytochemical method. In the brain, 5-HT immunostaining cells were localized in the raphen nuclear area of brain stem and in the ependyma cell area of the ventriclus tertius of diencephalon. These immunostaining cells were round or oval. The cells usually possess processes which were filled with immunoreactive substance. Some of the processes were contact with the processes of other cells. A few 5-HT positive reactive nerve fibers were observed in the brain stem and the diencephalon. The density of 5-HT immunostaining cells in the digestive tubes were the highest in the pylorus, fundus, cardia of gaster, and moderate in the esophagus and duodenum and the lowest in the large intestine and the small intestine. The density of 5-HT immunostaining cells in the digestive tubes were higher in nonhibernant toads than in hibernant toads. By the statistical method, the difference of the density between the two sorts of toads were notable (P less than 0.05). The 5-HT immunostaining cells were visualized to distribute between the epithelium cells of the mucosa or the epithelium cells of gland. These positive cells usually had one or more processes which contained 5-HT immunoreactive substance. Some were reached into lumen surface of the gland or intestine. Some were extended into lamina propria through the basal membrane. These results indicate that the 5-HT immunostaining cells in digestive tubes could release 5-HT by both endocrine and exocrine ways.     

Ontogeny of immunoreactive glicentin in the human gastrointestinal tract and endocrine pancreas

The gestational time of appearance and distribution of immunoreactive glicentin was compared to that of immunoreactive glucagon in the gastrointestinal tract and endocrine pancreas of human fetuses, aged between 5 and 24 weeks, by an indirect immunoperoxidase method. With the glicentin antiserum No. R 64, the first immunoreactive cells were detected at the 10th week of gestation in the oxyntic mucosa and proximal small intestine, at the 8th week in the ileum and at the 12th week in the colon. In the endocrine pancreas, the first immunoreactive cells were observed as early as 8 weeks within the walls of the primitive pancreatic ductules. At a more advanced stage of development (12 weeks), they were found interspersed among the islet cell clusters and still later (16 weeks) inside the recognizable islets of Langerhans. With the glucagon antiserum No. GB 5667, no immunoreactive cells were demonstrated in the gastrointestinal tract whatever the age of the fetuses. In the endocrine pancreas, the first immunoreactive cells were observed at the 8th week of gestation in the pancreatic parenchyma. The distribution of glucagon-containing cells in the pancreas was similar to that of glicentin immunoreactivity throughout ontogenesis. In the pancreatic islets of one 18-week-old human fetus, the study of consecutive semithin sections treated by both antisera showed that the same cells were labelled. The significance of these findings concerning the role of glicentin as a glucagon precursor is discussed.     

A light-microscopic immunocytochemical study of the endocrine pancreas in the Australian brush-tailed possum (Trichosurus vulpecula).

The endocrine pancreas of the Australian brush-tailed possum (Trichosurus vulpecula) was investigated by means of immunocytochemistry using the avidin-biotin-peroxidase technique. This was a light microscopic study using this established technique. Serial paraffin sections were stained individually with primary antibodies for glucagon, insulin, somatostatin, and pancreatic polypeptide (PP), showing the same islet. Cells immunoreactive to glucagon, insulin, somatostatin and PP were found in endocrine islets. PP cells appear to be scattered amidst the exocrine portion also. Insulin immunoreactive cells were located in the central region of islet, glucagon in the periphery, somatostatin in periphery and had elongated processes. PP cells were more sparse and located both in the periphery of islet and amidst the exocrine tissue. These results can then be related to a similar study in the same marsupial, but using the immunofluorescence technique and to studies in other marsupials such as grey kangaroo (Macropus fuliginosus) fat-tailed dunnart (Sminthopsis crasicaudata) and the American opossum (Didelphis virginiana). These investigations are part of a study in Australian mammals.     

Immunocytochemical study of tyrosine hydroxylase and dopamine β-hydroxylase immunoreactivities in the rat pancreas

An immunohistochemical and immunoelectron microscopic study was used to demonstrate tyrosine hydroxylase (TH) and dopamine -hydroxylase (DBH) immunoreactivities in the rat pancreas. Small TH immunoreactive cells were found in close contact with large TH immunonegative ganglion cells among the exocrine glands and were occasionally found in some islets. Some of these TH immunoreactive cells were also DBH immunopositive. The immunoreaction product was seen diffusely in the cytoplasm and in the granule cores of TH immunoreactive cells. All intra-pancreatic ganglion cells were immunoreactive for DBH, but not for TH. The TH immunoreactive cells were identified as small intensely fluorescent (SIF) cells due to their localization and morphological characteristics and showed no insulin, glucagon, somatostatin or pancreatic polypeptide immunoreactivities. These results indicate that SIF cells may release dopamine or noradrenaline to adequate stimuli while the intra-pancreatic ganglion cells with only DBH may not synthesize catecholamines in a normal biosynthetic pathway. TH immunoreactive nerve bundles without varicosities and fibers with varicosities, associated or unassociated with blood vessels, were found in both the exocrine and endocrine pancreas. Close apposition of TH immunoreactive nerve fibers to the smooth muscle and endothelial cells of the blood vessels was observed. A close apposition between TH immunoreactive nerve fibers and exocrine acinar cells and islet endocrine cells was sometimes found in the pancreas. The immunoreaction product was seen diffusely in the axoplasm and in the granular vesicles of the immunoreactive nerve fibers. Since no TH immunoreactive ganglion cells were present in the rat pancreas, the present study suggests that noradrenergic nerve fibers in the pancreas may be extrinsic in origin, and may exert an effect on the regulation of blood flow and on the secretory acitivity of the acinar cells, duct cells and endocrine cells.