Immunocytological Study Research Articles

Delayed muscle soreness and cytoskeletal alterations: an immunocytological study in man.

Delayed muscle soreness in man is known to be attributed to a disorganization of the myofibrils. Our aim in the present study was to investigate whether the intermediate filament system is affected as well. Biopsies were taken from the right m. vastus lateralis of six males (aged 23 +/- 3 years) suffering from severe post-exercise soreness of their thigh muscles. Intermediate filament protein was visualized by indirect immunofluorescence microscopy using an antibody specific to desmin. A typical finding in biopsies taken 3 days after exercise was abundant longitudinal "desmin" extensions. Moreover, strongly autofluorescent granules were observed beneath the sarcolemma. It is concluded that muscles subjected to unusual exercise contain myofibrillar and cytoskeletal disturbances accompanied by increased lysosomal activity.

Immunocytological study of the distribution of C-cells calcitonin in the thyroid gland of the normal adult gerbil.

The distribution of C-cells in the thyroid gland of the gerbil (Meriones inguiculatus) was studied on serial sections in the cranio-caudal direction. C-cells were visualized by immunofluorescence with an anti-calcitonin immune serum. A very heterogeneous distribution was found. C-cells first appear in the posterio-interior position. Then they progressively occupy the whole posterior part and finally are confined to the posterio-exterior position. No C-cells at all were detected in the upper and lower poles. According to the species, there is a wide variation in the relative distribution of C-cells in the thyroid gland. Our results add to the information available about the distribution of C-cells and its bearing on phylogenic evolution.

Organ culture of the anterior pituitary in a synthetic medium. An immunocytochemical study.

Anterior pituitaries removed from human foetuses and rat neonates were maintained in organ culture after using three different media. A comparative immunocytological study of the different cell-types was made in the three media used. In the first medium containing foetal calf serum the different cell-types were present but their frequency in the cultured explants depended upon their representation in the tissue at the beginning of the culture. In the medium without any adjunction, the most characteristic feature was the shrinkage of the cultured tissue. In the third medium in which foetal calf serum was replaced by insulin the evolution of the explants was the same as in the first medium. All the cell-types were represented with a particular development of gonadotrophic cells. In conclusion it seemed that the third synthetic medium can be used in order to study anterior pituitary cells in vitro.

Comparative Study in vivo and in vitro of the Differentiation of Immunoreactive Corticotropic Cells in Fetal Rat Anterior Pituitary

In order to study the mechanisms of the differentiation of adenohypophysial corticotropic cells, an immuno-cytological study was performed in fetal rat anterior pituitary in vivo and in vitro with antisera against beta-(1-24) and alpha-(17-39) ACTH and beta-LPH, alpha- and beta-endorphins. In vivo, these cells appeared at 16 days of gestation without any difference in the timing of appearance of the two immunoreactivities. The same immunoreactivity was also detected in adenohypophysial primordia explanted from 12 to 15 days of gestation and maintained in organ culture until 21 days by using either medium containing fetal calf serum or medium containing insulin and transferrin instead of fetal calf serum. These immunoreactive cells were first detected in the different experimental primordia after a minimal period of culture, corresponding to a final equivalent of 16 days as in vivo. However, the mean cytoplasmic area of immunoreactive cells increased in relation to the day of explantation whatever the duration of culture. These data suggest: (1) the nature of culture medium used in this study has no influence on the differentiation of the corticotropic cells; (2) this cell type seems to be committed precociously (before day 12) by one or several substances of unknown origin; (3) the normal development seems to require the presence of factors (before day 14) whose nature and origin remain to be elucidated.

Immunocytological study of the chronology of pituitary cytogenesis in the domestic pig (Sus scrofa) with special reference to the functioning of the hypothalamo-pituitary-gonadal axis.

This paper reports an immunofluorescent study of the pituitary in the fetal pig (Sus scrofa). Fifteen antisera were used against most of the hormones present in the pituitary. Five types of hypophysial endocrine cells were observed in the anterior and intermediate lobes. We determined the sequential appearance of these various cell types in the fetus. The first hormones found at 33 days were ACTH, beta-MSH, beta- and gamma-LPH and alpha- and beta-endorphin; alpha-MSH appeared at 40 days and STH at 45 days. The glycoprotein hormones, LH (45 days), TSH (50 days) and FSH (60 days), appeared between 45 and 60 days. The density and staining of the gonadotropes increased up to 80 days, at which time they reached values similar to those of the adult. Prolactin was not found until 80 days. An anti-LHRH antiserum was used to study LHRH neuron differentiation between 30 and 70 days of pregnancy. The first immunoreactive perikaryons were found at 40 days but the immunoreactive fibers did not reach the median eminence until about 60 days. However, we observed differentiated capillary loops in the palisade layer of the median eminence only in the 70-day fetus. These results when compared with actual data on the differentiation of the reproductive function in the pig fetus permitted us to define an overall pattern of the differentiation and functioning of the fetal neuroendocrine hypothalamo-pituitary-gonadal system in the porcine species. This pattern includes, (i) autodifferentiation and autofunctioning of the gonads and (ii) autodifferentiation of the pituitary with (iii) later assumption by the hypothalamus followed by a phase during which the whole reproductive system functions.

Monoclonal antibodies (O1 to O4) to oligodendrocyte cell surfaces: An immunocytological study in the central nervous system

Four monoclonal antibodies are characterized that have been obtained from a fusion of mouse myeloma P3-NS1/1-Ag4-1 with spleen cells from BALB/c mice immunized with white matter from bovine corpus callosum. The corresponding antigens (O antigens) are designated O1, O2, O3, and O4. The localization of these antigens was investigated by indirect immunofluorescence in cultures of early postnatal mouse cerebellum, cerebrum, spinal cord, optic nerve, and retina. When tested on live cultures none of the O antibodies reacted with the surface of astrocytes, neurons, or fibroblasts, however, all are positive on the surface of oligodendrocytes. The identity of these cells was determined by double-immunolabeling experiments with indpendent cell-type-specific antigenic markers (glial fibrillary acidic protein, tetanus toxin receptors, fibronectin, and galactocerebroside). Antigen O1 is exclusively expressed on galactocerebroside-positive cells, whereas O2, O3, and O4 are expressed on additional cells that are negative for any of the markers tested. None of the O antigens is expressed on the surface of cultured retinal cells. In fresh-frozen sections of adult mouse cerebellum all O antigens are detectable in white matter tracts and in vesicular structures of the granular layer. O2 and O3 antigens are in addition detectable in GFA protein-positive radial fibers in the molecular layer. In fixed cerebellar cultures, where intracellular antigens are accessible, O1, O2, and O3 antibodies label astrocytes in a GFA protein-like pattern. O antigens are expressed in mouse, rat, chicken, and human central nervous systems. O antibodies belong to the IgM immunoglobulin subclass and have been used in complement-dependent cytotoxic elimination of cerebellar oligodendrocytes in culture. At limiting antibody dilutions all processes of oligodendrocytes are preferably lysed over cell bodies.

Immunocytological study on the differentiation of chick and quail adenohypophysis epithelial rudiments, grafted or cultivated in vitro: evidence for polypeptidic hormones.

Epithelial rudiments of adenohypohysis were removed from chick and quail embryos between days 3 and 5 of development. Chick rudiments were grafted for 11--13 days onto the chorioallantoic membrane of decapitated chick embryo hosts. Quail rudiments were cultivated in vitro for 6 days. Both grafted and cultivated Rathke's pouches differentiated into adenohypophyseal tissue. The adenohypophyseal tissue cultured on chorio-allantoic membrane exhibited cells reacting with the following immune sera: anti-beta-(1--24)ACTH, anti-alpha-(17--39)-ACTH, anti-alpha-endorphin, anti-beta-endorphin and anti-beta-LPH, which also gave a positive reaction when applied to adenohypophysis of corresponding age which had differentiated in situ. In situ, corticotrophs were located exclusively in the cephalic lobe of adenohypophysis. Therefore, the differentiation of corticotrophs in the whole graft, i.e., from both cephalic and caudal lobes of Rathke's pouch, showed that the cells of the caudal lobe, or at least some of them, were uncommitted when the rudiment was removed. In vitro, tissue derived from Rathke's pouch contained cells reacting with antibodies to beta-(1--24)-ACTH, alpha-(17--39)-ACTH, and beta-LPH, as did adenohypophysis from quail embryos of corresponding age (9--10 days), differentiated in situ. The differentiation of quail Rathke's pouch in vitro corroborates that differentiation can occur without influence from hypothalamus and, moreover, shows that at least some kinds of cells can differentiate without influence exerted by any other encephalic factors, and in the absence of mesenchyme. The question arises whether fibroblastic cells derived from Rathke's pouch cells act as feeder-cells and/or secrete some factors promoting differentiation.

The Gonadotropic Cell in Parr, Precocious Parr Male and Smolt of the Atlantic Salmon, Salmo salar. An Immunocytological, Light- and Electron Microscopical Study

Lindahl, K. 1980. The gonadotropic cell in parr, precocious parr male and smolt of the Atlantic salmon, Salmo salar. An immunocytological, light- and electron microscopical study. (Department of Zoology, University of Stockholm, Sweden.) — Acta zool. (Stockh.) 61(2): 117–125. Only one GTH cell type was identified in the pars distalis of the parr, smolt and precocious parr male. On a fine structural basis four phases with transitional forms of the GTH cell were recognized. In parr and smolt the GTH cells have numerous secretory granules and their cytology substantiate that they are in a storing phase with a possibly low release rate. On the contrary, in the precocious parr male all phases of the GTH cell are present. The great number of active cells and frequent degranulation and vacuolization indicate a high activity and release level.

Comparative distribution of somatostatin, LH-RH, neurophysin, and α-endorphin in the rainbow trout: An immunocytological study

Using immunofluorescence, evidence of a somatostatin (SRIF)-like antigen has been found in the brain and digestive tract of rainbow trout. In the diencephalon, periventricular SRIF immunoreactive hypendymocytes are located in the region dorsal to the nucleus preopticus (NPO). SRIF immunoreactive perikarya are concentrated anterior to the NPO in the nucleus preopticus periventricularis, scattered in small cells in the nucleus lateralis tuberis (NLT) pars anterior, and in a few cells located in an unnamed nucleus in the dorsomedial hypothalamus. In the pituitary SRIF immunoreactive material is located in the neurohypophysial tissue in the proximal pars distalis. In the gut, SRIF cells have been found in the endocrine pancreas and in the gastric mucosa. Comparatively, material immunoreactive for luteinizing hormone-releasing hormone has the same distribution in the pituitary as SRIF, whereas neurophysin immunoreactivity was found in only the neurophysial tissue of the neurointermediate lobe. A few cells reacting with anti-α-endorphin were seen in the NLT in the pituitary stalk region. All the pars intermedia cells in the neurointermediate lobe react with anti-α-endorphin.

Étude immuno-cytologique des neurones hypothalamiquesàLH-RH chez le foetus humain

The immunocytological study of LH-RH producing neurons was carried out on 3 newborns and 14 fetuses (from 10 to 36 weeks of age). Perikarya and fibers which were immunoreactive to anti-LH-RH IS were revealed by IF or IE in all the hypothalamus beginning with the 13th week. Important variations in neurons staining may translate “physiological” differences in their LH-RH charge but can be also the result of the diverse technical conditions. In three 16-week-old female fetuses, the large number of neurons (more than 150 per hypothalamus) permitted a good topographical and morphological study of them. They are scattered in vast areas of the anterior hypothalamus (lamina terminalis (LT) and septum), mediobasal and premammillary hypothalamus. The fibers which are particularly immunoreactive in semi-thin sections form a large hypothalamo-infundibular contingent in the posterior lip of the ME where they give rise to collaterals that terminate in contact with the capillaries of the mantelplexus, this taking place both before and after the apparition of the intra-eminential loops at the 16th week. Numerous in the LT, they terminate around the deep capillaries of the vascular organ or in contact with the ependymal epithelium. Some extra hypophyseal fibers go towards the epithalamus and the mesencephalon. To conclude, very early, in the human fetus the peptidergic LH-RH system resembles that described in adult primates; its role in the maturation and control of the gonadotropic cells is evoked.