The influence of thyrotropin and growth hormone on the thyroid gland in the hereditary dwarf mouse: a morphometric study.

Snell-type dwarf mice were injected with TSH, GH, or both hormones together for 6 days. GH induced an increase in body weight but not in the weight of the thyroid gland itself; on the contrary, TSH caused an increase in the weight of the thyroid but no increase in body weight. After TSH injection, the relative volume of the thyroid parenchyme was enhanced by 45% compared to that in untreated dwarf mice, and the radius of the follicles and follicular lumina increased by 50% and 48%, respectively. The major effect of TSH was an increase in cellular volume (+93%), and the mean number of cells in the average follicle was doubled, without a reduction in the number of follicles. GH had almost the same effect as TSH on the relative volume of the parenchyme and caused the radius of follicles and of the follicular lumina to increase by 61% and 69%, respectively. However, GH did not influence cellular volume. Its primary effect was to stimulate cellular division (cells were increased about 5 times in the average follicle) and to reduce the number of follicles. The nucleo-cytoplasmic ratio increased with GH but decreased with TSH. T4 serum levels increased to a much lesser extent with GH than with TSH, while normal values were obtained with both hormones together. At a morphological level, the combined administration of TSH and GH produced the same qualitative effects as separate administration, inducing an increase in cell volume and number which was less than the sum of the effects of each hormone administered separately.

Flow cytometric detection of erythropoietic cytotoxicity in mouse bone marrow.

Erythroblast proliferation and maturation in bone marrow are the processes leading to the formation of polychromatic erythrocytes (PE) and normochromatic erythrocytes (NE), respectively. PE contain RNA but no DNA, and can therefore be distinguished both from NE (which lack both RNA and DNA) and from nucleated cells (which contain both DNA and RNA). Cytotoxic agents that induce impairment of the maturation process change the PE:NE ratio. We have developed a simple and rapid method of determining the PE:NE ratio, based on flow cytometric analysis of formaldehyde-fixed, acridine orange (AO)-stained cells. The effects of cyclophosphamide (CP), mitomycin C (MMC), and vincristine (VC) were tested and the PE:NE ratio was evaluated over 7 days of treatment. In this study we monitored the kinetics of these compounds and were able to demonstrate both a time- and a dose-dependent effect. We detected a difference between the effects of the alkylating agents tested and those induced by the spindle inhibitor tested. Flow cytometry of fixed bone marrow samples stained with AO provides more information, better and more rapid statistical analysis, than conventional microscopic methods for counting the PE:NE ratio.

Morphometry of nuclear pore complexes in thyroid cells during hyperplasia and involution.

Nuclear pore complexes were analyzed in freeze-fractured replicas of thyroid follicular cells of C3H mice in different physiological states. Thyroid stimulation induced a rapid and simultaneous increase of the nuclear surface and volume and of the total number of pore complexes. The numerical density (Na) of pore complexes increased at the 6th day of stimulation, but after that time the proportion of cells with an increased Na was always higher than the proportion of 3H-labelled nuclei. During thyroid involution, all the nuclear parameters, including the Na, returned to normal values. These results indicate that the total number of pore complexes and their Na are correlated with the cellular activity rather than with the cell cycle. They also suggest that 2 different mechanisms are involved in the generation of pore complexes: first, an addition of new membranes with a low density of pore complexes; later, a formation of new pore complexes in preexisting membranes. However, during involution, parts of the nuclear membranes and pore complexes in the remaining parts disappear synchronously. In freeze-fractured thyroid nuclear membranes, 2 neighbouring pore complexes were always separated by a distance of 105 nm. Clusters of pore complexes were not observed. A comparison of the distances between pore complexes and between randomly generated points never showed any significant differences indicating that pore complexes were randomly distributed.

The thymus of Nu/+ mice.

At one month, the thymus weight of NMRI mice heterozygous for the Nu gene was lower than that of the wild type. However, the thymic structure was normal and no significant anomalies at the ultrastructurallevel could be found either in epithelial cells or in thymocytes. Stereologic data showed that the loss of weight was only due to a decrease of the lymphoid population, whereas the absolute number of epithelial cells was unmodified. Several hypotheses that could explain the reduced number of thymocytes were discussed.

Relationship between ciliary rootlets and smooth endoplasmic reticulum.

In ciliated cells of thymic cysts in Nude mice, ciliary rootlets are constantly and closely related to smooth endoplasmic reticulum and clear vesicles. This special association suggests that this structure does not play only an anchoring role but must be involved in the general metabolism of the cilium.

Ultrastructure of the cilia of thymic cysts in "nude" mice.

The ciliary ultrastructure is studied in the ciliated cells of the thymic cysts of the "nude" mouse. The cilium is made up of two segments: the intracellular basal body and the extracellular cilium shaft. The basal body extends in a rootlet and demonstrates three annexes: basal foot, anchoring system and tonofibrillar web. The rootlet, composed of one central and nine peripheral chambers, is closely and constantly associated with smooth endoplasmic reticulum. The classical periodicity of the basal foot can be explained by its internal structure. The anchoring system binds the upper part of the basal body to the cell membrane. The cilium shaft is divided into four segments: implantation base, proximal segment, intermediate segment and tip. The details of particular structures in the implantation base and in the proximal segment are given on transverse and longitudinal sections. In the intermediate part, a peculiar desmosome-like structure, binds some doublets with the ciliary membrane. At the tip level, A tubules end up in an opaque plate and membranes are covered with several intracated cristae.

Origin of necklace particles in thymic ciliating cells.

The formation of ciliary necklaces during ciliogenesis in a thymic cyst was observed in freeze-etched replicas. The necklaces first appear as clusters of particles arranged in concentric circles on a flat area of the cell membrane. As soon as the cilium begins to grow, the particles move to the periphery.

Nude mouse embryo: ectodermal nature of the primordial thymic defect.

Sterile female homozygous nude mice were rendered fertile through a graft of thymic epithelium from heterozygous littermates 3 days after birth. Homozygous nude embryos were compared with embryos age (8-18 days). In nude mice, the embryological anomaly is noticeable from the 11th day of gestation and essentially consists of a failure of the ectoderm from the 3rd branchial cleft to proliferate and differentiate into a cervical vesicle. As a result, the endoderm from the 3rd pouch degenerates into cystic formations instead of differentiating.

Epithelial cell proliferation in thymic hyperplasia induced by triiodothyronine.

A significant hyperplasia of the thymus was induced in mice, treated with triiodothyronine during the first month of life. Stereological data showed that, in both treated and control mice, mononucleate epithelial cells were four times more numerous in the medulla that in the cortex. After triiodothyronine treatment, their absolute number in both cortex and medulla increased two-fold. The number of thymic epithelial cells could thus be regulated by thyroid hormones. The cortical volume in treated mice was also twice that of controls but medullar volume showed and increase of only fifty percent. Cortical epithelial cells increased at the same rate of the cortex volume by medullary epithelial cells grew more rapidly. In fact the medullary volume enlargement could be be explained mainly by the growth of the epithelium. Medullary lymphocytes did thus not preliferate in the same way as cortical lymphocytes after thyroid hormone administration. The recently described multinucleate epithelial cells were not modified in number and were thus insensitive to thyroid hormones.

Ultrastructure of the thyroid in dwarf mice.

The thyroid gland of Snell's dwarf mice consists of small follicles with flattened epithelium, partly differentiated cords and undifferentiated masses. Many adipocytes are found. The ultimobranchial cysts are well developed. Parafollicular cells are normal. In the partly organized cords, microfollicular cells are some small follicles limited by two or three cells are seen. The presence of these structures led us to think that they represent the first stages of normal folliculogenesis, described as the fusion of two adjacent unicellular microfollicles. Their further growth is the result of the coalescing of small adjacent follicles or of cellular multiplication. The presence of undifferentiated masses and partly differentiated cords, in dwarf mice, seems due to a developmental arrest rather than to aberrant development. This disorder of organogenesis is ascribed mainly to a lack of pituitary growth hormone.

Thyroid gland in dwarf mice: a stereological study.

Stereological methods were used to compare thyroids of dwarf mice and of their heterozygote littermates. In the thyroid of dwarf mice unorganized cellular masses, adipous tissue and ultimobranchial cysts are abundant. Follicles are small and their distribution function is unimodal. The number of cells per follicle is considerably lowered if compared with the normal. In control mice the distribution function of thyroid follicles is bimodal. These data show that origin of the thyroid anomaly in dwarf mice is due to a drastic diminution of cell divisions, probably resulting from the lack of growth hormone.

The influence of fixation procedure, embedding medium and section thickness on morphometric data in thyroid gland.

In this study, the effects of fixation procedures, embedding medium and section thickness on stereological measurements of normal thyroid were analysed. The following conclusions were drawn: A) the use of a single section for the analysis of a lobe is sufficient if this section is located in the central part of the lobe. B) fixation and embedding with glutaraldehyde-Epon leads to a larger shrinkage than Bouin-paraplast, but the difference between the two procedures is not significant. C) osmium post-fixation reduces the shrinkage induced by glutaraldehyde and lowers the axial deformation produced by sectioning. D) Bouin's fixative and paraplast embedding induce considerable shrinkage of the interstitial tissue. The shrinkage obtained with glutaraldehyde-Epon is less. However, it is still not known whether this difference is due to the fixative, or to the embedding procedure or to both. E) only in glutaraldehyde and osmium-fixed material, embedded in Epon, can follicles and colloids be assumed to be spherical in shape without significant errors.

Ultrastructural aspects of crystal-like inclusions in a case of IgA plasma cell proliferation.

A case of apparently benign plasma cell proliferation is described. Needle-like paracrystalline inclusions were observed in numerous plasma cells outside the cisternae of the rough endoplasmic reticulum. They had a dense protein core surrounded by a layer of carbohydrate. They were found only in plasma cells secreting IgA-kappa immunoglobulin, but did not react with labelled specific antisera. A small amount of paraprotein IgA-kappa was found in the serum but no Bence-Jones protein could be detected in the urine. These observations suggest that these plasma cells synthesize normal IgA immunoglobulin and accumulate abnormal protein in the form of paracrystalline inclusions.