Basic and applied histochemistry最新文献

Neuroendocrine cells (NE) occurring in the pulmonary epithelium of the fishes Polypterus delhezi and P. ornatipinnis are studied by electron microscopy and by immunostaining for serotonin which is often present in such cells in the mammalian lung. With the electron microscopy NE are found to occur single, resting upon the basement membrane and forming a narrow cytoplasmic extension towards the air lumen. They contain dense-cored vesicles of 80-165 nm which form exocytotic profiles at the level of the basal membrane. An immunoreactivity for serotonin is demonstrated for the first time in the NE of these species. The role of this mediator may involve a paracrine or endocrine function as postulated for the respiratory neuroendocrine mammalian cells. NE of the species studied are considered similar to those found within the wall of lung airways in mammals and submammalian vertebrates. Although much immunocytochemical investigations remain to be executed, they may also be included in the APUD (or DNES) cell system.

Langerhans cells (LCs) were shown up in normal cervical tissue obtained from 32 women whose age ranged from 25 to 68 years, using immunohistological methods. LCs were detected in metaplastic and native squamous epithelium of the cervix; they were positive to Dako-LC and OKDR antibodies and those located in the basal-suprabasal epithelial layers were also OKT6-positive. The density of the LCs was higher during the proliferative phase of the menstrual cycle. The investigation into the lymphocytes present in the stroma and in the squamous epithelium showed a population of T-lymphocytes identified as predominantly T-cytotoxic/suppressor cells, sometimes in contact with LCs. The significance of these findings is discussed.

The applicability of acetic anhydride (AA) in dimethyl sulfoxide (DMSO) for the oxidation of polysaccharide and their subsequent visualization with thiocarbohydrazide (TCH) and silver proteinate (SP) was evaluated on LR White-embedded thick and ultrathin liver sections. The results of these studies indicated that AA-DMSO-TCH-SP reaction is chemically specific on LR White-embedded tissues and that it offers distinct advantages for the localization of minute glycogen aggregates.

The mechanism of postlarval fish myotomal growth was investigated in trout (Salmo gairdneri) by means of morphometric and cytofluorometric analysis. The mechanism by which new fibres are added during postlarval growth (hyperplasia) is not fully understood. In histological cross sections these new fibres have a small diameter which give the muscle a "mosaic" appearance. One hypothesis suggested that they could be derived from the proliferative activity of satellite cells. DNA cytofluorometric analysis of nuclei suspensions obtained from trout white myotomal muscle during different developmental stages (eleutherembyronic; alevin; yearling and adult) showed a consistently low S-cytometric phase during all stage in which myofibres of small diameters were present. The percentage of such small fibres, determined by morphometric analysis, suggested that satellite cells are the proliferative population. In fact, their percentages, as determined by morphometric analysis in histological section, bear a linear relationship with the S-cytometric phase percent nuclei (R = 0.927). Only in adults (67 cm in size) there was a significant decrease in the S-cytometric phase. At this stage, in histological sections, the myotomal muscle no longer had a "mosaic" appearance because of the disappearance of the small fibres. It may, therefore, be supposed that in the cm 67 adult specimens, the proliferative population is entering the G0 phase. It is known, in fact, that muscle growth proceeds only by fibre hypertrophy in trout longer than 70 cm in length (Stickland, 1983).

The cytochemical pattern and the quantitative assay of DHFR on Daucus carota cell lines sensitive and resistant to methotrexate (MTX) are described. Cytoplasmatic enzyme activity is evidenced as granules of formazan, final reaction product, in the majority of the cells. DHFR activity appears low in Daucus carota cells used as control while in carrot cells resistant to MTX is very high, an observation which parallels previous biochemical studies. These results are supported by quantitative data of DHFR content in a random cell population. A possible correlation between overproduction of DHFR and gene amplification in MTX-resistant cells is discussed but the solution of this problem will depend on the availability of a dhfr plant probe.

The thyroid tumours and background goiterous and adenomatous lesions induced in rats with diisopropanolnitrosamine (DIPN) plus methylthiouracil (MTU), and regenerative thyroid tissues after wounding were studied by lectin histochemistry. Ten weeks after cessation of the carcinogen treatments, carcinomas invading the surrounding tissues and blood vessels (13/20) and papillary micronodules (11/20) were formed in the thyroid tissues. In general, the carcinoma lesion was solitary, and the papillary micronodules were multiple in a single thyroid gland. Among the lectins tested, Maclura pomifera (MPA) and Solanum tuberosum (STA) showed specific binding with both carcinoma and papillary micronodule lesions, but not with the background goiterous and adenomatous lesions and regenerative thyroid tissues. The former both lesions showed higher labelling indices with BUdR or 3H-thymidine and poorer thyroglobulin accumulation than the latters, thereby indicating their enhanced proliferative capability and depressed potency of cyto-differentiation. The common cytological and histochemical properties of carcinoma lesions and papillary micronodules allow us to regard the latter as pre-invading carcinoma lesions. The lectins MPA and STA may be, therefore, used as the specific markers of malignancy in rat thyroid carcinogenesis.

Enzyme and carbohydrate histochemical methods were used to study the secretory activities and secretion properties of the eccrine tubular glands in the foot pad of the cat. The activity spectra of the different oxidative and hydrolytic enzymes investigated indicate high metabolic rates within the secretory epithelium. Additionally the enzyme reactions emphasize a double innervation of the glands by cholinergic and adrenergic nerve fibres. The carbohydrate histochemical differentiation reveals mostly neutral and very few acidic glycoproteins in the secretory cells and the secretion, respectively. Clear (basal) cells contain distinct amounts of glycogen, and dark (superficial) cells show neutral glycoproteins, which reveal after PO-lectin-DAB procedures the following saccharide residues: alpha-D-mannose, alpha-D-galactose, N-acetyl-alpha-D-glucosamine, alpha-L-fucose, beta-D-galactose-D-N-galactosamine, beta-D-galactose, and sialic acid. The results obtained confirm the view that the normal biological functions of the eccrine glands of the feline foot pad are to improve the frictional capacities of the paw and to leave typical scent marks.

Prostate Specific Antigen (PSA) is regarded as a specific marker of prostatic epithelium and has never been detected by immunocytochemistry in extra-prostatic tissues. The casual finding of a strong positivity for polyclonal antisera to PSA in a sweat gland carcinoma prompted a study on a series of skin adnexial and breast specimens (normal and neoplastic). Normal axillary and perineal apocrine sweat glands, some apocrine foci in fibrocystic breast disease and two sweat gland and two breast apocrine carcinomas were stained by several PSA antisera; a recently introduced monoclonal to PSA, however, was unreactive. These observations cast doubt on the specificity of PSA for prostatic epithelium, especially when polyclonal antisera are employed. Immunocytochemical reactions obtained with PSA, in the investigation of skin, lesions must be interpreted with caution and confirmed if necessary with monoclonals to PSA and with PAP.

Lectin-binding patterns were examined in epithelial walls of 65 jaw cysts (30 post-operative maxillary cysts: POMCs, 20 radicular and 15 follicular cysts), and characteristic lectin staining for each kind of jaw cysts is presented. Between squamous and columnar epithelia, the staining intensity of WGA, Con A and UEA-I was not different, but SBA bound more remarkably to squamous than to columnar epithelia. In both epithelia the outer layers did react more strongly with the lectins examined. Concerning odontogenic cysts, the lectin-binding affinities of outer and intermediate layer cells were nearly the same in both follicular and radicular cysts. Basal cells of radicular cyst walls were however, more markedly positive for lectin binding than of follicular cysts. Furthermore, basal cells of keratinized (RKSE 60 keratin-positive) epithelium were inferior to those of non-keratinized linings in the bindings. Lectin-binding patterns of metaplastic squamose epithelia of POMCs which were positive for RGE53-keratin (principally columnar epithelium-specific keratin) were similar to originally squamous linings of odontogenic cysts. Columnar linings of unusual radicular cysts were positively stained with SBA. By these results, lectin-binding sugar residues of the epithelium seem to be related to the epithelial morphology.