小鼠生后不同发育阶段附睾组织中Crb3的表达

应用HE染色法和免疫荧光组织化学技术分别对小鼠生后不同发育阶段附睾组织结构及Crb3在不同发育时期附睾组织中的定位表达进行了研究。结果显示,附睾在4周龄时上皮为2层～3层的假复层纤毛柱状细胞,顶端纤毛细长;6周龄上皮细胞层数减少至1层～2层;8周龄附睾管上皮厚度增至最大,上皮细胞呈单层高柱状;12周龄后附睾管上皮开始变薄,呈单层柱状。免疫荧光染色结果显示,Crb3蛋白主要分布在附睾上皮柱状细胞的胞膜,在精子尾部有微弱的表达,这提示了Crb3不仅与附睾上皮细胞的极性建立和维持有关,而且对精子活力和血-附睾屏障的形成也可能具有潜在的影响。     

Cultivation of corneal epithelial cell sheets on canine amniotic membrane

Objective To develop and assess canine corneal epithelial cell sheets cultivated from limbal stem cells on amniotic membrane. Procedures Canine corneal limbal segments were obtained from six beagle dogs. Cryopreserved denuded amniotic membranes (obtained from Miniature Dachshund and Cavalier King Charles Spaniel breeds) from which the epithelial cells were removed were used as scaffolds. The limbal segments were cultured on these amniotic membranes with 3T3 feeder cells for 2 weeks. The harvested corneal epithelial cell sheets were stained with H&E for histologic analysis. The harvested sheets were analyzed immunohistochemically using a corneal epithelium‐specific marker keratin 3(K3) and putative stem cell markers ABCG2, p63, and vimentin. Results Cultivated cells from the corneal limbal tissues reached confluency in 7–8 days. The cultivated cells adhered to the denuded amniotic membrane and formed a sheet. The cultivated cell sheet was transparent and consisted of five to eight layers. K3 was observed in all layers and ABCG2, p63, and vimentin were notably present in the basal layer of the cultivated canine epithelium by immunofluorescence. Conclusions Canine corneal epithelial cells were successfully cultivated on the canine amniotic membrane. The cultivated epithelial sheets contained putative stem cells in the basal layer and had a stratified epithelium.     

Structure and Steroidogenesis of the Placenta in the Antarctic Minke Whale (Balaenoptera bonaerensis)

There are few reports describing the structure and function of the whale placenta with the advance of pregnancy. In this study, therefore, the placenta and nonpregnant uterus of the Antarctic minke whale were observed morphologically and immunohistochemically. Placentas and nonpregnant uteri were collected from the 15th, 16th and 18th Japanese Whale Research Programme with Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. In the macro- and microscopic observations, the placenta of the Antarctic minke whale was a diffuse and epitheliochorial placenta. The chorion was interdigitated to the endometrium by primary, secondary and tertiary villi, which contained no specialized trophoblast cells such as binucleate cells, and the interdigitation became complicated with the progress of gestation. Furthermore, fetal and maternal blood vessels indented deeply into the trophoblast cells and endometrial epithelium respectively with fetal growth. The minke whale placenta showed a fold-like shape as opposed to a finger-like shape. In both nonpregnant and pregnant uteri, many uterine glands were distributed. The uterine glands in the superficial layer of the pregnant endometrium had a wide lumen and large epithelial cells as compared with those in the deep layer. On the other hand, in the nonpregnant endometrium, the uterine glands had a narrower lumen and smaller epithelial cells than in the pregnant endometrium. In immunohistochemical detection, immunoreactivity for P450scc was detected in most trophoblast cells, but not in nonpregnant uteri, suggesting that trophoblast epithelial cells synthesized and secreted the sex steroid hormones and/or their precursors to maintain the pregnancy in the Antarctic minke whale.     

Pathogenesis of experimental bovine mastitis following a small inoculum of Escherichia coli

Mastitis was produced in four quarters of two lactating cows by the inoculation of 50 or 200 viable Escherichia coli. Changes were investigated by light microscopy and scanning electron microscopy. After 10 hours the changes were confined to the superficial layer of the epithelium of the teat and lactiferous sinuses; single cells or small groups of cells were damaged and were being extruded from the tissue. By 14 hours there was extensive necrosis and sloughing of the epithelial cells which did not extend beyond the basement membrane, and an intense inflammatory response associated with the epithelial damage. The somatic cells in the milk, mainly polymorphonuclear leucocytes, increased 40 to 250 times and strongly inhibited the survival of E coli. Epithelial damage appeared first in the ventral portions of the gland and then extended through the lactiferous sinus to the large ducts and secreting tissue. This pattern of damage together with the failure to observe bacteria attached to the epithelia was consistent with the production of diffusible toxins by the organisms.     

Functional indispensability of fibronectin associated on the mesenchymal cell surface during the early period of feather development in the chick embryo in vitro

To clarify the role of fibronectin (FN) during the early period of feather development, reconstituted skin consisting of intact epithelium and isolated mesenchymal cells from embryonal chick skin was used. In early feather development, FN was localized around mesenchymal cells of the dermal condensation. Isolated mesenchymal cells had associated with FN on their surfaces. FN on the cell surface dissociated following EDTA treatment, and EDTA‐treated cells re‐associated with exogenous FN. The intact epithelium also bound to exogenous FN at the placode. When FN‐associated or FN‐reassociated mesenchymal cells were used, the reconstituted skin formed feather rudiments only at the positions where the epithelial placode existed originally, and the locality of tenascin transferred from the placode to the mesenchyme during the period of feather bud formation. However, in reconstituted skin using FN‐dissociated mesenchymal cells, feather rudiments did not form. Additionally, the epithelial placodes disappeared, and tenascin was distributed uniformly on the surface of the epithelium and not localized in the mesenchyme. These findings suggest that FN associated on the surfaces of mesenchymal cells maintains the functions of mesenchymal cells as dermal condensation and mediates epithelial‐mesenchymal interactions during the early period of feather development. The results also suggest that reconstituted skin is a useful tool for functional studies on the extracellular matrix.     

The Anatomy of the Cloacal Bursa (Bursa of Fabricius) in the Helmeted Guinea Fowl (Numida meleagris galeata)

The cloacal bursa (bursa of Fabricius) in the guinea fowls appeared either as an oval blind sac with a short thick stalk in one group or had a pointed cranial blind end with a slightly bulging middle part that was followed by a thick caudal stalk in the other group. Both groups of bursae originated from the proctodeal wall of the cloaca and were placed dorsal to the rectum. The average length of the bursa was 18 mm while the average width at die mid section was 15 mm. The internal surface showed about 12 – 14 primary folds. Histologically, the outline of the bursa was well established by day 18 of incubation. The primary folds had also been formed. Lymphocytes had already been encountered within the framework of the bursa at this day. The epithelium bordering the tunica propria was composed principally of two layers of cuboidal cells. Epithelial buds had also formed and some were already detached from the epithelial lining. The blood vessels present were positioned just beneath the outer covering. At day 19 of incubation, most of the epithelial buds had two layers of cells arranged in a circumscribed manner while a few had three layers of cells. Blood vessels had increased in number and were deeper placed inside the bursa than previously. At day 20, the cells of the upper layer of the epithelium were dorsoventrally flattened and stained paler than the cells of the lower layer. It was possible to distinguish the cortex from the medulla and the basement lining between both zones was distinct. Tiny vesicles within the cytoplasm of the epithelial cells at the mucosa and follicles were observed. Macrophges were also observed within the gland. At day 21, blood vessels were observed in the cortex of the follicles. The maximum number of primary folds (14) had been formed. At day 22, serveral follicles had severed connections with the mucosal epithelium. The mucosal lining had dropped to a single layer of cells in some areas. Goblet cells were observed amongst the mucosal cells. A plasma cell had first appeared. By day 25, dead cells had increased quite in number and there was also an increase in number of medium and small-sized lymphocytes within the gland. By day 26, the upper layer of the surface epithelium was composed primarily of tall columnar cells with numerous large vacuoles. Macrophages had suddenly increased within the thin interfollicu-lar spaces and most of them were crowded internally with various sizes of debris. By day one post-hatch, each fold was completely filled with follicles that were separated by thin connective tissue strands.     

Co‐culture of Buffalo Preantral Follicles with Different Somatic Cells

The effect of co‐culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250–450 μm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 μm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co‐cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (μm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co‐cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co‐culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.     

Pathogenesis of Staphylococcus aureus mastitis: bacteriologic, histologic, and ultrastructural pathologic findings

Three lactating cows were experimentally infected with Staphylococcus aureus ATCC 29740 (Newbould 305 strain). Cows were euthanatized 2 to 216 hours after inoculation. Bacteriologic and microscopic examinations showed that S aureus attached to epithelial cells of the mammary gland in vivo. The histopathologic changes observed were progressive swelling, vacuolar degeneration of epithelial layers, and multiple foci of epithelial erosions and ulcers throughout the ductal system. The cellular response of the infected glands was demonstrated by a rapid increase in the number of somatic cells in the secretion and by accumulation of neutrophils below, within, and on the epithelium of the teat and lactiferous sinuses. The inflammatory response did not prevent infection nor subsequent pathologic changes in the inoculated glands.     

Aural carcinoma with chondroid metaplasia at metastatic sites in a dog

A case of aural carcinoma with chondroid metaplasia at metastatic foci in an 8-year-old male pug is described. Multiple metastases in both lungs and the right submandibular, parotid, retropharyngeal, cervical and prescapular lymph nodes were detected. Histologically, the skin of the right ear canal appeared to be diffusely infiltrated by cords and nests of neoplastic epithelial cells, showing multifocal contiguity with the overlying hyperplastic squamous epithelium. Most of the carcinomatous cells were arranged in a glandular-like pattern, with formation of lumens containing epithelial cells attached to the peripheral cell layer by elongated intercellular bridges. Scattered foci of keratinization with central accumulations of compact, laminated keratin were also observed, and histochemical stains failed to detect mucinous secretory material. Even though histological and histochemical findings were compatible with a diagnosis of acantholytic squamous cell carcinoma, CAM5.2 immunostaining was detectable in the majority, although not all, neoplastic cells, confirming a diagnosis of poorly differentiated ceruminous gland carcinoma. Pulmonary metastatic nodules revealed multifocal areas of cartilaginous metaplasia with apparent transition of carcinomatous cells to chondroid cells, showing nuclear atypia and focal cytokeratin immunostaining. Carcinomatous cells surrounding chondroid areas also revealed focal vimentin and S100 immunoreactivity. Histological evidence of transition between the two components, as well as the presence of intermediate cells displaying both epithelial and mesenchymal immunohistochemical features, strongly indicated a final diagnosis of carcinosarcoma, in which chondrosarcomatous elements were derived from carcinoma cells.     

贝氏莫尼茨绦虫感染绵羊对小肠局部黏膜淋巴组织的影响

为了研究贝氏莫尼茨绦虫自然感染绵羊对小肠黏膜免疫组织的影响,分别从宏观、微观及亚微观水平对自然感染贝氏莫尼茨绦虫的成年绵羊(感染组)肠道进行了细致地观察,并与正常成年绵羊(正常组)进行了比较.结果显示,感染组肠道所见虫体平均长度为1.5m,头节主要吸附在空肠淋巴集结分布丰富的部位,一般寄生数量为1～2条.眼观,虫体寄生部位黏膜增厚,表面有大量灰白色黏液附着,其间可见点状出血.镜下,局部黏膜上皮脱落,而在完整的黏膜上皮处,其上皮细胞、上皮内淋巴细胞、杯状细胞的数量都明显增多;固有层内毛细血管充血,淋巴细胞、浆细胞、弥散淋巴组织以及肠腺杯状细胞均有不同程度的增生,头节寄生处部分肠腺坏死;黏膜下层淋巴小结、淋巴集结显著增生,部分增生凸入固有层形成新的圆顶区;固有层与黏膜下层以及黏膜肌层可见大量嗜酸性粒细胞浸润.扫描电镜下,感染组肠黏膜上皮脱落;贝氏莫尼茨绦虫头节呈椭球状,有4个吸盘,无顶突,小沟,表面覆盖一层致密的微绒毛.研究结果表明,肠黏膜增厚,主要是局部黏膜免疫相关细胞在寄生虫虫体表面覆盖的微绒毛的不断刺激下,机体抗感染自身组织增生所致.成年绵羊对抗贝氏莫尼茨绦虫的感染可能是通过黏膜免疫相关组织增生来加强局部免疫力而实现的.     

Estudio Ontogenico Comparativo de la Lamina epitelial de los Compartimentos Gastricos no Glandulares en Ovinos Merinos

A total of 74 embryos and fetuses were used in a comparative analysis of the epithelium of the non-glandular stomach compartments of merino sheep during development. The mechanical protection showed by the tegumentary epithelium in the superficial layers of the rumen, reticulum and omasum is supported by a buffer system of neutral mucopolysaccharides secreted by the deeper strata. Neutral mucopolysaccharides first appeared in epithelial cells at 46 days of fetal life. Acid mucopolysaccharides, mucins, and mucoid compounds were not detected. Growth curves and formulas were constructed for the epithelial layers.     

Histomorphometric analysis of the irides of dogs, camels, buffalos and donkeys

The present study was carried out to investigate the morphological and histomorphometrical characters of irides in dogs, camels, buffalos, and donkeys. The findings of the study revealed that, morphologically, the irides were consisted of an anterior border layer, a middle layer of connective tissue stroma and a posterior layer of pigmented epithelium. Interestingly, the anterior borders of all investigated animals were not enveloped by a distinct epithelial or endothelial lining, but on contrary, the posterior border was covered by pigmented epithelium. The constrictor and dilator iridial muscles were well developed in dogs, weakly developed in donkeys, and with an intermediate position in camels and buffalos. Morphometric analysis revealed significant species differences in the mean total thickness of the iris and its different layers. In addition significant differences were also found between the ratio of the means of different layers to the total thickness of the iris at the pupillary, middle and ciliary borders. In conclusion, these variations might be related to the different lifestyles and visual behaviour of the investigated animals.     

鲫鱼和斑马鱼视网膜结构的比较组织学研究

为探讨鲫鱼(Carassius auratus)和斑马鱼(Brachydanio rerio)视网膜组织结构与其生活环境的适应关系,对这2种鱼视网膜各层厚度、3个核层的胞核层数进行了测量,并对数据进行了统计分析。结果表明:鲫鱼和斑马鱼视网膜均由4层细胞构成,在光镜下共分为10层。鲫鱼视网膜平均厚度196.57μm,斑马鱼视网膜平均厚度167.57μm。斑马鱼的视网膜内核层的细胞数目比外核层多,而鲫鱼的视网膜外核层细胞数目多于内核层数目。鲫鱼和斑马鱼这2种鱼类视网膜结构的差异,与其各自的生活环境相适应。     

Histology and morphometry of the testes of adult domestic cats (Felis catus)

Testicles of 30 mongrel cats were analyzed histologically and morphometrically, divided into three groups: G1 (1-2 years old), G2 (over 2 and up to 4 years old) and G3 (over 4 and up to 6 years old). After orchiectomy and histopathology, the morphometric parameters studied were: thickness of the tunica albuginea (72 μm) and seminiferous epithelium (77.19 μm), perimeter (53.81; 90.57 μm), (54.80; 101.07 μm); area (174.23; 494.55 μm(2)), (176.68; 629.70 μm(2)); maximum diameter (14.94; 28.02 μm), (14.76; 31.66 μm); minimum diameter (13.25; 21.92 μm), (13.30; 24.52 μm); and shape factor (index for regularity of the format) (1.36; 1.36), (1.39; 1.35) of the nucleus and cytoplasm of spermatogonia and Leydig cells, respectively. The results can be used for comparative studies and contribute knowledge concerning the height of the seminiferous epithelium, thickness of the tunica albuginea and size of spermatogonia and Leydig cells.     

八点黑獭兔不同发育时期子宫组织结构变化的研究

观察母兔妊娠20、26、29天和15、30、60、90日龄八点黑獭兔的子宫组织,以探讨八点黑獭兔子宫的形态学发育特点。结果表明：獭兔的子宫随着年龄的增长．其内膜的粘膜上皮由假复层柱状纤毛上皮逐渐分化成紧密规则的单层柱状上皮;固有层在15～30日龄之间出现．内含大量间质细胞;妊娠期固有层发生水肿和充血;子宫肌层在幼龄时分层不明显,各层肌纤维互相交织,随平滑肌细胞分裂增生,3月龄时肌层出现明显的内环、外纵两层,中间形成一层环状排列的血管：獭兔出生后其子宫内膜腺由子宫内膜上皮细胞内陷形成,随后腺上皮细胞经有丝分裂增生,15日龄幼兔已形成单管子宫腺。该研究结果可为兔的解剖学、生殖生理学增添新的内容,也为进一步研究兔生殖系统疾病的治疗与预防提供参考资料。     

Acute airsacculitis in turkeys inoculated with Pasteurella multocida

Thirty female turkeys, inoculated into the caudal thoracic air sacs with Pasteurella multocida were examined from 0 to 6 hours post-inoculation (PI). The air sac reacted rapidly and intensely with exudation of heterophils. Circulating leukocyte and thrombocyte numbers remained normal except for an absolute lymphopenia by 6 hours PI. P. multocida was initially isolated from blood at 3 hours PI. Total cell counts increased markedly in air sac lavage fluids by 1.5 hours PI and continued to increase until 6 hours PI. Heterophils predominated in lavage fluids (greater than 94%), with macrophages comprising the remaining cells. Microscopically occasional heterophils were present within air sac blood vessels and perivascularly by 0.5 hour PI. They became more numerous by 1.5 and 3 hours PI when transepithelial migration into the air sac lumen was seen. By 6 hours PI, there was diffuse, severe swelling of air sac epithelium and mesothelium, and bacteria were located in air sac interstitium. Ultrastructurally, endothelial and air sac epithelial cells were swollen and vacuolated Interdigitating processes of air sac epithelial cells were separated. These results indicate that air sacs can be the portal of entry for P. multocida into the systemic circulation, probably via damaged air sac epithelium.     

Interspecies comparative morphological evaluation of the corneal epithelial stem cell niche: a pilot observational study

BackgroundThe corneal and limbal morphology relevant to corneal epithelial maintenance in ten different species was examined using histological methods.ObjectivesThe presence of a Bowman’s layer, limbal epithelial cell, and superficial stromal morphology was examined in the following species to evaluate the differences in corneal thickness and epithelium: Java sparrows, frogs, macaws, spoonbills, red pandas, penguins, horses, Dobermans, orangutans, and humans.MethodsCorneal sections (4 µm) were obtained from ten ocular globes from three different animal classes: Aves, Amphibia, and Mammalia. All sections were stained with hematoxylin and eosin and periodic acid-Schiff reaction. After microscopy, all stained slides were photographed and analyzed.ResultsSignificant morphological differences in the corneal and limbal epithelia and their underlying stroma between species were observed. The number of corneal epithelial cell layers and the overall corneal epithelial thickness varied significantly among the species. The presence of a Bowman’s layer was only observed in primates (orangutans and humans). Presumed supranuclear melanin caps were noted in four species (orangutans, macaws, red pandas, and horses) in the limbal basal epithelial layer (putative site of corneal epithelial stem cells). The melanin granules covered the apex of the cell nucleus.ConclusionsSupranuclear melanin capping has been described as a process within the epidermis to reduce the concentration of ultraviolet-induced DNA photoproducts. Similarly, there may be a relationship between limbal stem cell melanin capping as a protective mechanism against ultra-violet radiation.     

Fine Structure of the Choriocapillaris, Bruch''s Membrane and Retinal Epithelium of the Cow

The fine structure of the choriocapillaris, Bruch's membrane and retinal epithelium was investigated in both the tapetal and non-tapetal fundus of the bovine eye. In ail locations the retinal epithelium consists of a single layer of cuboidal cells. The epithelial cells are joined laterally by apically located tight junctions and throughout the retina display numerous basal infoldings and fine apical processes which enclose rod outer segments. All retinal epithelial cells are rich in smooth endoplasmic reticulum and mitochondria and contain phagosomes. Although not as abundant, profiles of rough endoplasmic reticulum and polysomes are also noted in all locations. In non-tapetal areas, melano-somes are numerous whereas over the central tapetum fibrosum they are absent. The absence of melanosomes over a functional tapetum is to be expected. While lysosomes are present throughout the epithelial layer, over the tapetal region they appear to be more numerous. The apparent increase in lysosomal numbers in this location may indicate an enhanced shedding of outer segment material over the tapetum. Although some retinal epithelial cells are modified to accomodate a tapetum lucidum their morphology is basically similar throughout the retina and probably indicates that ail regions of the retinal epithelium are capable of the normal functions of this indispensible retinal layer. The choriocapillaris is heavily fenestrated on the border facing the retina and overlying the tapetum also displays fenestrae on its choroidal edge. Bruch's membrane ( complexus basalis ) is pentalaminate throughout the retina and is slightly thicker in the posterior fundus.     

Expression and adhesive ability of gicerin, a cell adhesion molecule, in the pock lesions of chorioallantoic membranes infected with an avian poxvirus.

The expression and adhesive activities of gicerin, a cell adhesion protein, in the pock lesions on chicken chorioallantoic membranes (CAM) infected with an avian poxvirus were studied. In normal CAMs, gicerin was found on the flattened epithelial cells, and neurite outgrowth factor (NOF) was in the basement membrane. However, in the pock lesions on infected CAMs, gicerin was overexpressed on the cell membranes of hyperplastic epithelial cells forming thick epithelial layers. Neurite outgrowth factor was also found mainly in the basement membrane, but occasionally showed aberrant expression among hyperplastic cells. In vitro analyses, using the dissociated cells from pock lesions, demonstrated that an anti-gicerin polyclonal antibody inhibit cell aggregation activity and cell adhesion to NOF. These results suggest that gicerin might promote the cell-cell and cell-extracellular matrix protein bindings of the hyperplastic epithelial cells by its homophilic and heterophilic adhesive activities, and contribute to pock formation on the infected CAMs.     

Evaluation of commercially available antibodies to cytokeratin intermediate filaments and laminin in normal cat pinna.

The pattern of distribution of cytokeratin (CK) intermediate filaments can be used to characterize subsets of epithelial tissues. The purpose of the study was to examine the CK expression of feline pinna skin. Six normal feline pinnae were routinely processed in formalin. An immunohistochemical method was used to stain the pinnae with 8 commercially available anti-human CK antibodies (Abs) (PKK1, CAM 5.2, UCD 10/11, 35BH11, 34BE12, AE1/AE3, MAK 6, A575) and an anti-human laminin Ab. All the CK Abs selectively localized to epithelium except 35BH11, which did not react with any part of the pinna. Some epithelial subsets were identified by their unique staining pattern with CK Abs. Basal cells but not suprabasal cells of the epidermis stained with PKK1; basal but not lumenal cells of apocrine glands stained with 34BE12. Apocrine glands stained with all CK Abs except 35BH11. All epithelial structures were stained with A575. Basal lamina of epithelial and mesenchymal tissues was clearly identified by the anti-laminin Ab. The results indicate that in cat pinna some commercially available anti-human CK Abs selectively stain subsets of epithelium and adnexa. PKK1, 34BE12, and A575 were the CK Abs with the most consistent staining patterns, the other Abs stained more variably from pinna to pinna. The pattern of epithelial and adnexal staining was similar but not identical to that reported for humans.