极北鲵消化道6种内分泌细胞的免疫组织化学研究

应用卵白素-生物素-过氧化物酶复合物(ABC)免疫组织化学法,对极北鲵(Salamandrella keyserlingii)消化道5-羟色胺(5-HT)、胃泌素(Gas)、生长抑素(SS)、胰高血糖素(Glu)、人胰多肽(PP)和P-物质(SP)6种内分泌细胞的分布进行了研究。文中采用Duncan多重比较的方法,对消化道内分泌细胞的分布密度进行统计学分析。结果显示,5-羟色胺细胞在消化道各部位均有分布,胃体(2.80±0.70)和十二指肠(2.60±0.75)分布密度最高,幽门最低(0.85±0.67);胃泌素细胞仅分布于小肠,十二指肠(1.85±0.75)密度最高;生长抑素细胞分布于食管、胃体、幽门、十二指肠和回肠各部位,且幽门(2.25±0.64)分布密度最高;胰高血糖素细胞只见于胃贲门部和胃体,人胰多肽细胞和P-物质细胞在整个消化道均未检测到。各种内分泌细胞的形态多样,呈圆形、椭圆形、锥体形和梭形等。极北鲵消化道内分泌细胞的这种密度分布和形态特点与其他两栖类动物及黑龙江产地的极北鲵相比,既有共性又有差异,其原因可能与物种、食性和栖息地环境等因素有关。     

极北鲵肠内5-羟色胺、胃泌素和生长抑素内分泌细胞胚后发育

目的 观察极北鲵胚后发育期间肠内5-羟色胺(5-HT)、胃泌素(gastrin,GAS)和生长抑素(somatostatin,SS)3种内分泌细胞的分布、形态和数量变化。方法 应用亲和素-生物素复合物(avidin-biotin complex,ABC)法免疫组织化学技术,对38-46期健康极北鲵肠内5-羟色胺(5-HT)、胃泌素(GAS)和生长抑素(SS)细胞进行染色,并统计分析不同时期极北鲵肠内3种内分泌细胞的密度。结果 5-HT免疫反应阳性细胞和GAS免疫反应阳性细胞最先在41期检出,而SS免疫反应阳性细胞在42期才检测到。5-HT细胞在41期最少,46期最多,呈年龄相关的递增趋势;GAS细胞在43期最少,44期最多,呈先减后增变化趋势;SS细胞在43期密度高于其它各时期。5-HT细胞最先分布在上皮基部,GAS细胞和SS细胞最先在肠上皮之间检测到。细胞形态多样,有椭圆形、三角形和锥体形等,但以锥体形为主。结论 极北鲵胚后不同发育时期肠内3种内分泌细胞的形态与其个体生理活动的改变相适应,且在数量变化上有其种属特异性。     

8周有氧运动对肥胖大鼠消化道嗜银 细胞形态及分布密度的影响

为了研究有氧运动对肥胖大鼠（Rattus norvegicus）消化道嗜银细胞形态及分布密度的影响,本实验采用Grimelius银染法观察8周运动组（n = 9）与对照组（n = 9）肥胖大鼠消化道嗜银细胞形态及分布密度。结果显示,大鼠消化道各部位均有嗜银细胞分布;两组大鼠消化道嗜银细胞形态上无差异,均以圆形、椭圆形、锥体形、梭形为主;两组大鼠消化道嗜银细胞分布密度高峰均位于胃体,而低谷有所不同,对照组大鼠消化道嗜银细胞的分布密度低谷位于食管、贲门,运动组大鼠位于食管、贲门、空肠、回肠、直肠;两组相比,食管和直肠两部位分布密度差异不显著（P > 0.05）,其余各部位均有差异,且运动组大鼠贲门、胃体、盲肠、结肠嗜银细胞分布密度极显著低于对照组（P < 0.01）,幽门、空肠嗜银细胞分布密度极显著高于对照组（P < 0.01）,运动组十二指肠、回肠嗜银细胞分布密度显著高于对照组（P < 0.05）。两组动物嗜银细胞分泌密度的这种改变与动物机体所处不同生理状态以及消化道各部位功能有关。     

中国黄羽鹌鹑消化道嗜银细胞胚后发育的分布及形态学观察

采用Grimelius银染法,对中国黄羽鹌鹑(Coturnix aponica)消化道嗜银细胞胚后发育的分布规律和形态结构进行了观察。结果显示,口腔、食管、嗉囊和泄殖腔中未发现嗜银细胞,其余部位均有不同数量的分布,其分布呈波浪形,大多数日龄段在腺胃和结直肠中存在2个分布高峰,回肠次之,十二指肠、空肠、盲肠较少,随着日龄增加,不同部位嗜银细胞数量均先增加,后减少,在100 d时达高峰。嗜银细胞主要分布在腺胃腺叶内细胞之间、肠黏膜上皮细胞之间及固有层内,形态多呈圆形、椭圆形、锥形及梭形等。结合嗜银细胞形态与功能间的联系,发现消化道内广泛分布着4种类型的嗜银细胞。我们认为中国黄羽鹌鹑消化道的嗜银细胞具有内分泌、外分泌及旁分泌3种功能,其分布特点可能与动物的食性及生活环境有关。     

猫儿山两种有尾两栖类消化道嗜银细胞比较观察

为探究有尾两栖类消化道内分泌细胞的特点,以分布于猫儿山保护区的猫儿山小鲵（Hynobius maoershanensis）和瑶山肥螈（Pachytriton inexpectatus）为研究对象,采用Grimelius银染法,对二者消化道嗜银细胞的形态特征、分布区域与分布密度进行了比较研究,为有尾两栖类消化道比较组织学和内分泌学研究积累基础资料,并为两栖类消化生理学研究提供形态学数据基础。结果显示,两种动物从食道至直肠均有嗜银细胞分布,分布型都呈波浪形,均在食道处分布密度（个/视野）最高,猫儿山小鲵为2.00 ± 0.69,瑶山肥螈为3.42 ± 0.31,在猫儿山小鲵回肠（1.45 ± 0.50）和瑶山肥螈在直肠（1.63 ± 0.97）分布密度最低。瑶山肥螈消化道各部位嗜银细胞的分布密度都高于猫儿山小鲵,两种动物嗜银细胞分布密度在食道、贲门、十二指肠和回肠中差异显著,但在胃体、幽门和直肠中差异不显著。对同一物种消化道不同部位嗜银细胞进行比较,猫儿山小鲵及瑶山肥螈的嗜银细胞分布密度高峰均出现在食道,其中,猫儿山小鲵食道与十二指肠、回肠、直肠差异显著（P < 0.05）,瑶山肥螈食道与其他各部位差异显著（P < 0.05）。嗜银细胞形状多样,有圆形、椭圆形、锥形和梭形等,可分为闭合型和开放型细胞。猫儿山小鲵生活环境多为沼泽地,富含淤泥、落叶,喜食蛙类蝌蚪、蚯蚓等个体较小且易消化的食物,瑶山肥螈生活环境为石块粗砂较多的山溪,以虾、蟹、螺等为食,这两种动物消化道嗜银细胞的形态特征和分布密度各具自身特点,可能与其生境及食性等因素有关。     

褐家鼠消化道嗜银细胞的分布及形态学观察

应用Grimelius银染法观察了褐家鼠Arattus norvegicus消化道嗜银细胞的分布密度及形态.结果 显示,在褐家鼠的消化道除食管未见嗜银细胞外,其它部位均有嗜银细胞的分布,其分布密度为胃贲门部最高,回肠部最低.嗜银细胞多分布于消化道粘膜上皮细胞之间或腺泡上皮之间,有时可见于上皮基部和固有膜内.嗜银细胞形态多样,主要以锥体形为主,其次还有圆形、梭形和椭圆形.根据其嗜银细胞形态,认为褐家鼠消化道内嗜银细胞具有内、外分泌、旁分泌3种功能.     

循环饥饿投喂对大鼠消化道嗜银细胞 形态及分布的影响

为了研究循环饥饿投喂对大鼠(Rattus norvegicus)消化道内嗜银细胞的分布密度及形态功能的影响,本实验采用Grimelius银染法观察和测定循环饥饿投喂(饥饿1 d,投喂1 d,周期为14 d)期间大鼠消化道嗜银细胞形态功能及密度分布。结果表明,实验组(即循环饥饿投喂组)和对照组(正常喂食组)大鼠消化道嗜银细胞除食管外均有分布,形态上对照组嗜银细胞以椭圆形和锥体形为主,实验组嗜银细胞则主要以锥体形为主;两组大鼠消化道嗜银细胞的分布密度高峰都位于胃,密度低谷对照组位于空肠、盲肠、回肠和直肠,实验组位于空肠到直肠各段;实验组大鼠消化道嗜银细胞分布密度在贲门和幽门部极显著低于对照组(P 0.01),结肠显著低于对照组(P 0.05),说明循环饥饿投喂会明显改变消化道嗜银细胞的形态并降低细胞数量,这可能与内分泌细胞的功能改变有关。     

北方狭口蛙消化道嗜银细胞的分布与形态

应用Grimelius银染法观察北方狭口蛙消化道嗜银细胞的形态学特点,并统计分析嗜银细胞的分布密度。结果表明,消化道嗜银细胞可分为开放型细胞和闭合型细胞,两者的比值变化范围为0.20～0.49,从食管至直肠都以闭合型细胞为主。分布密度食管和胃体最高,空肠最低。北方狭口蛙消化道嗜银细胞的形态学特点和分布密度有自身的独特性,这可能与其善于穴居掘土的生活习性有关。     

凹耳蛙消化道组织学和嗜银细胞形态观察

为了揭示凹耳蛙(Odorrana tormota)消化道的基本特征,运用石蜡切片法和龙桂开银浸法对凹耳蛙消化道组织学结构及嗜银细胞的形态与分布密度进行了观察。结果显示:①凹耳蛙的胃壁具明显的纵行皱襞和胃小凹,胃腺发达,小肠可分为十二指肠和回肠,杯状细胞分散在十二指肠上皮细胞之间,十二指肠中未见十二指肠腺分布。②凹耳蛙嗜银细胞见于消化道全长,呈毛笔头样、锥体形、梭形、椭圆形和长条形等;幽门腺上皮和十二指肠绒毛上皮中的嗜银细胞具指向腺泡腔或肠腔的突起,提示其可能具有腔分泌的功能。嗜银细胞的分布密度胃幽门部最高,十二指肠和胃体其次,食道最低。据此认为胃既是凹耳蛙的主要消化器官,也是消化道中主要的内分泌器官;十二指肠是凹耳蛙消化道中的主要吸收部位,同时也具有内分泌功能;消化道嗜银细胞具有内分泌的功能,还可能具有腔分泌的功能。     

Phylogenetic relationships of two Salamandrella species as revealed by mitochondrial DNA and allozyme variation (Amphibia: Caudata: Hynobiidae)

We conducted molecular phylogenetic analyses to confirm taxonomic relationships and to delimit distributional ranges of Siberian salamanders, Salamandrella keyserlingii and Salamandrella schrenckii, and to elucidate the origin of the isolated population of this species complex on Hokkaido, Japan. Phylogenetic trees constructed by MP, NJ, ML, and Bayesian methods, using complete sequences of mitochondrial cytochrome b genes, all indicated monophyly of Salamandrella and of each of the two species. Identical relationships were found on UPGMA, NJ, and CONTML trees derived from electrophoretic analysis of variation in 18 inferred allozyme loci. Populations from Hokkaido and northeastern China proved to be S. keyserlingii, while populations from Khabarovsk and Lazovsky are S. schrenckii. Genetic differentiations of S. keyserlingii within Sakhalin, and between Sakhalin and Hokkaido, are substantial. The Hokkaido population is hypothesized to have been isolated on the island since early Pleistocene, much earlier than isolation of sympatric anuran populations from their Sakhalin relatives. In contrast, the continental populations of S. keyserlingii are only slightly differentiated from some Sakhalin populations, and are thought to have expanded their ranges in the late Pleistocene.     

Phylogeography of the Siberian newt (Salamandrella keyserlingii) by mitochondrial DNA sequence analysis

Differentiation of geographical populations of the Siberian newt Salamandrella keyserlingii throughout the species range was analyzed using a fragment of the cytochrome b gene. The population of the Primorye region (Russian Far East) is separated to the greatest extent; the Japanese and South Kuril populations are the next most separate. These populations are possibly subspecies. Geographical differentiation of populations in the Siberian part of the species range is lower, lacks a clinal variation, and is irregular. The molecular variation of S. keyserlingii supports the hypothesis that several primary vicarious refugia of pre-Pleistocene differentiation of a common ancestor of Salamandrella occurred in the southeastern part of its current distribution range and that northern and western regions were gradually colonized via repeated steps of expansion and retreat in the Siberian part of the modem species range.     

扬子鳄消化道嗜银细胞的分布及形态学观察

该文用龙桂开浸银法对扬子鳄消化道嗜银细胞的分布及形态进行了观察。结果表明：嗜银细胞分布于整个消化道中,从食道到直肠。其中,十二指肠和回、直肠交接处密度很高,胃体及直肠很低。嗜银细胞形态多样。食道嗜银细胞位于上皮基部和固有膜中,呈椭圆形或不规则形。胃嗜银细胞位于胃腺部,圆形或椭圆形,有的可见明显的胞突。肠嗜银细胞位于上皮细胞之间,呈长柱形、纺锤形、长颈瓶形或锤状。多数细胞两端有较长胞突,分别与固有膜     

鲤稚幼早期发育过程中粘液细胞的发生和变化

利用酸性条件下的阿新蓝染色和过碘酸－雪夫氏试剂反应相接合的方法（AB－PAS染色）,对鲤早期发育过程中粘液细胞的变化作了初步研究。结果表明,鲤的粘中液细胞在受精卵孵化前一天出现,密度随个体的而增大,成分随发育过程的进行而趋于复杂,鲤的粘液细胞可分为四种类型,Ⅰ型和Ⅱ型含单一成分,为幼稚型,Ⅲ型和Ⅳ型含复合成分,为成熟型,鲤粘液细胞在不同组织器官中的分布是不均匀的,其主要分布区是皮肤、口腔、鳃、消化道等部位。     

Comparative analysis of gastrin-immunoreactive and argyrophil cells in bony fish larvae

A comparative study was made of enteroendocrine cells in the larvae of bony fishes (Teleostei): carp (Cyprinus carpio L.), big head (Aristichyts nobilis Richardson), silver carp (Hypophthalmichthys molitrix Valenciennes), and atlantic salmon (salmo salar L.). Argyrophil cells demonstrated by the method after Grimelius were found in all examined fish species. They are located in the pyloric appendices, stomach and intestinal mucosa. Gastrin-immunoreactive cells demonstrated by the immunocytochemical method were located in the pyloric part of the stomach in all examined fishes, except for the atlantic salmon in which they were not identified. The first argyrophil cells were identified at the age of about 10 days, while gastrin-immunoreactive cells proved to appear 5-7 days later.     

Postnatal development of the brush cells in the common bile duct of the rat

Summary The postnatal development of brush cells in the distal segment of the common bile duct of the rat was examined with respect to cell number and immuno-reactivity for liver fatty acid-binding protein (L-FABP). The brush cells, distinguishable from the principal cells by scanning electron microscopy, first appeared in the common bile duct 4 weeks after birth. They showed a remarkable increase in number, with a sex difference in time, i.e., between 8 and 12 weeks in the male and between 10 and 14 weeks in the female. In both sexes, the frequency of brush cells reached approximately 30% of total epithelial cells by 16 weeks and remained constant until 40 weeks of age. Cells with positive immuno-reactivity for L-FABP first appeared in small numbers at 8 weeks. Immuno-electron microscopy revealed that all immunoreactive cells were brush cells. They increased in number gradually from 16 to 40 weeks with no sex difference. At 40 weeks, the immunoreactive cells reached approximately 7.5% of total epithelial cells, corresponding to one-fourth of the number of brush cells. These results indicate that the occurrence of the brush cell population in the common bile duct is a late event in the postnatal development of the rat and that its functional maturation progresses with aging.     

Appearance and distribution of fetal brain macrophages in mice

Summary A study on the localization of fetal and neonatal brain macrophages of mice from embryonic day 10 (E10) to postnatal day 21 (P21) was carried out immunohistochemically using a monoclonal antibody against a macrophage differentiation antigen (Mac-1) and the labeled avidin-biotin technique. In the central nervous system, the macrophages recognized first were mainly located in the choroid plexuses of the fourth and lateral ventricles at E14. Their number increased at E17–P3 and gradually decreased thereafter. In the cerebral parenchyma, a few macrophages appeared at E14 in the matrix cell layer. They were also detected in the migrating zone at E15, E17 and in the cortical plate at E19. Mapping of positive cells at the stage of neuroblast formation (E15, E17, E19) disclosed the precise distribution of cerebral macrophages. The macrophages that appeared first in the choroid plexuses at E15 may be derived from the subarachnoid vessels, which extend into the stroma of the choroid plexuses when the matrix cell layer invaginates into the lateral ventricle to form the choroid plexuses. Almost all of the macrophages recognized in the cerebral parenchyma disappeared at P9 when the cytoarchitecture seemed to be completed. In the cerebellum, which develops later than the cerebrum, macrophages appeared after birth and were located mainly in the internal granular layer. The brain macrophages always appeared in the regions where cell proliferation and brain remodeling are most active at each stage. These findings suggest that fetal and neonatal brain macrophages may play an important role in scavenging degenerated cells and cell debris during histogenesis of the central nervous system.     

Transient appearance of Ca-binding protein (spot 35-calbindin) in bronchial epithelial cells,thyroid parafollicular cells and thymic epithelial cells during the development of rats

Summary Tracheobronchial epithelium, thyroid organ, thymus, of the developing rats were examined by immunohistochemistry using anti-spot 35 calbindin-antiserum. At E 14, weak to moderate immunoreactivity for spot 35-calbindin was detected in the airway epithelia of the distal half of the trachea and the extrapulmonary bronchus. The immunoreactive cells increased in intensity at E 16–E 21, but decreased markedly after birth. These cells were non-ciliated cells and comprised a majority of the epithelial cells especially in the ventral/cartilaginous portion of the airway. They were characterized by microvilli, vacuoles, granular and agranular endoplasmic reticulum. Typical ciliated cells, which were much less numerous than the immunopositive non-ciliated cells, were immunonegative. In thyroid gland, calbindin-immunoreactive cells first appeared at E 18. They increased in number at E 20-P 1 and decreased gradually after P 7. These cells were the parafollicular cells characterized by numerous secretory granules and situated in close proximity to the basal surface of the follicular cells. In the thymus, immunoreactive cells appeared in the thymic medulla at E 20. They increased in number at P 1, but decreased gradually after P 7. They were stellate in shape and had vesicles, vacuoles, intermediate filaments and represented a subpopulation of thymic reticular epithelial cells. Such a transient appearance of spot 35-calbindin in these cells suggests that this protein may be involved in the regulation of differentiation or may be involved in the process of secretion during the limited developmental period.     

Transient appearance of Ca-binding protein (spot 35-calbindin) in bronchial epithelial cells, thyroid parafollicular cells and thymic epithelial cells during the development of rats.

Tracheobronchial epithelium, thyroid organ, thymus, of the developing rats were examined by immunohistochemistry using anti-spot 35 calbindin-antiserum. At E 14, weak to moderate immunoreactivity for spot 35-calbindin was detected in the airway epithelia of the distal half of the trachea and the extrapulmonary bronchus. The immunoreactive cells increased in intensity at E 16-E 21, but decreased markedly after birth. These cells were non-ciliated cells and comprised a majority of the epithelial cells especially in the ventral/cartilaginous portion of the airway. They were characterized by microvilli, vacuoles, granular and agranular endoplasmic reticulum. Typical ciliated cells, which were much less numerous than the immunopositive non-ciliated cells, were immunonegative. In thyroid gland, calbindin-immunoreactive cells first appeared at E 18. They increased in number at E 20-P 1 and decreased gradually after P 7. These cells were the parafollicular cells characterized by numerous secretory granules and situated in close proximity to the basal surface of the follicular cells. In the thymus, immunoreactive cells appeared in the thymic medulla at E 20. They increased in number at P 1, but decreased gradually after P 7. They were stellate in shape and had vesicles, vacuoles, intermediate filaments and represented a subpopulation of thymic reticular epithelial cells. Such a transient appearance of spot 35-calbindin in these cells suggests that this protein may be involved in the regulation of differentiation or may be involved in the process of secretion during the limited developmental period.