Anatomical Record最新文献

Enucleation is the last event in the development of a definitive erythroid line, and extruded nuclei are phagocytosed by macrophages. Both colchicine and cytochalasin have been known to exert a great influence on the enucleation process, but the relationship between enucleation and these agents has not yet been clearly revealed in vivo. Our aim was to clarify the significance of the enucleation in liver erythropoiesis and macrophage phagocytosis by colchicine and cytochalasin administration to embryonic mice. Pregnant mice were intraperitoneally injected with colchicine or cytochalasin at 13 days of gestation. Embryonic livers were removed at intervals of 3, 6 and 12 h after injection for processing for light and electron microscopy, and, to obtain three-dimensional morphology of erythroids at enucleation, computer-aided reconstructions were performed by light microscopy. Colchicine injections had cytolytic effects on hepatocytes and macrophages, and numerous erythroblasts were observed in the process of enucleation after colchicine injection. However, the extruding nuclei were irregularly shaped, and some erythroblasts at mitosis showed extreme peripheralization of their chromosomal masses and cell membrane constriction. Enucleation behavior could also be observed in immature erythroblasts. Liver macrophages engulfed extruded nuclei and erythroblasts in mitosis. Cytochalasin injections, on the other hand, had no significant effect on embryonic livers. The progress of erythroblast mitosis was clearly stopped by colchicine injection, and numerous erythroblasts at mitosis were extruding their nuclear compartment. Following colchicine injection, erythroid enucleation also took place in immature erythroblasts, and mitotic erythroids were phagocytosed. In enucleation, more attention should be paid to hematopoietic environmental factors than to hemopoietic cell factors.

Background: Maternal diabetes influences fetal pancreas development. As there are some controversial reports, we studied the morphometric changes of the fetal insular pancreas and insulin immunostain of beta cells as well as the proliferative activity of insular cells in 21-day-old fetuses from control, diabetic, and insulin-treated diabetic pregnant rats.

Methods: Streptozotocin was injected into 7-day-pregnant rats (controls were not injected). Some rats were either left untreated (diabetic) or injected with insulin. Animals were killed at 21 days of gestation. Fetal pancreas were fixed in toto for the morphometry and immunohistochemistry studies using anti-insulin, anti-Ki-67 and anti-proliferating cell nuclear antigen (PCNA) antibodies.

Results: Diabetic status was determined by measuring maternal and fetal serum glucose and insulin levels. The morphometric studies showed hyperplasia of the diabetic fetal insular tissue which had not been normalized by insulin therapy. Diabetes caused an increase of both insulin-positive and insulin-negative cells. The increase in insulin-positive cells was not corrected by insulin treatment, although the number of non-beta cells became normal. The nuclear area in beta cells increased in diabetic rats but was not corrected by insulin. The cytoplasmic area decreased in diabetic rats and was normalized by insulin administration. Diabetes increased the expression of the nuclear antigen Ki-67 in fetal insular pancreas, and insulin treatment returned it to the normal state.

Conclusions: Maternal diabetes leads to hyperstimulation of fetal beta cells, with increased proliferative activity. Insulin administration to the dams corrects some of the changes observed.

Background: The pituitary gland of the dog is different from all animals and is described as "typical" for mammals. How might this complex pituitary gland of the dog be formed in fetal life? The current study examined the fetal development of the complex and unique dog pituitary and the ontogeny of specific cell types in the pars distalis.

Methods: Adenohypophysis of the beagle, from 25 to 60 days of gestation and at 2 days of age, was studied by immunocytochemical and histological staining.

Results: At 25 days gestation, the primordium (Rathke's pouch) of the adenohypophysis began to form by an upward evagination from the epithelium of the primary oral cavity. At 38 days, the pituitary gland showed the same morphology as in adult dogs, being merely smaller. Five walls of Rathke's pouch (anterior wall [A], lateral walls [L], posterior wall [P], and upper wall [U] were found at 25 days, and by 38 days they had specialized into specific regions of the adenohypophysis through complex and unique processes. The pars intermedia was derived from the U and the dorsal area of the A. The pars tuberalis was derived from the dorsal area of the A. The pars distalis was formed by more singular processes: the peripheral areas of the pars distalis were first formed by A and P; then the ventral lumen of the extensive Rathke's lumen surrounded by these areas was filled up by proliferation of cells, although the dorsal lumen remains as Rathke's lumen after 38 days. The blood capillaries may play an important role in the development of parencymal cells in the Rathke's pouch during canine fetal life. At 30 days gestation, the first adrenocorticotropic hormone cells were found in the anterior- and posterior-ventral regions (derived from middle and ventral areas of the A and the P) of the pars distalis anlage, and blood capillaries invaded the parenchymal cells from the mesencyme surrounding the anlage. At 38 days, portal vessels without capillary loops in the median eminence had appeared, and growth hormone and luteinizing hormone cells appeared in the ventral areas of A and P in the pars distalis. By 52 days, when capillary loops were seen in the portal vessels in the median eminence, these types of cells spread through the whole pars distalis.

Conclusion: These areas in the epithelium of Rathke's pouch at 25 days may differentiate into specific regions of the pars distalis during subsequent fetal life, through complex processes that are characteristics to the canine species.

Background: The Class III beta-tubulin isotype (beta III) is expressed specifically in central and peripheral nervous system neurons at various stages of neuronal differentiation. We have shown previously that beta III is expressed in a differentiation-dependent manner in human neuroblastomas arising in the adrenal medulla and sympathetic chains (Katsetos et al., Clin Neuropathol 13:241-255, 1994). The neuronal distribution of beta III in the developing and mature human adrenal medullae is detailed in the companion article (Katsetos et al., 1998A).

Methods: We have compared the localization of the neuronal beta III to S-100 protein, a sustentacular cell marker, in 14 formalin-fixed, paraffin-embedded primary human pheochromocytomas of the adrenal medulla and 14 adrenocortical tumors (adenomas and carcinomas).

Results: In pheochromocytomas, beta III staining was present in all tumors, but the number of stained cells varied in the two neural neoplastic phenotypes. Although the majority of chromaffin-like cells were beta III-positive, there was a lack of beta III in one-third of the tumor cells. Compared to chromaffin-like phenotypes, neuronal (ganglion-like cells) were invariably beta III-positive. Stromal sustentacular cells, stromal fibroblasts, and tumor blood vessels were beta III-negative. Sustentacular cells in pheochromocytomas were S-100 protein-positive, but beta III-negative. Primary adrenocortical tumors were beta III-negative with the exception of rare beta III-positive cells demonstrated in one case.

Conclusions: The distribution of beta III in human pheochromocytomas of the adrenal gland is differentiation-dependent, closely recapitulating chromaffin cell and neuronal phenotypes of the normal adrenal medulla. Our findings indicate that beta III may be used as one of the adjuvant neural markers in the differential diagnosis of adrenal tumors, i.e., pheochromocytoma versus adrenocortical carcinoma. The occurrence of rare beta III-positive cells in cortical carcinomas is exceptional and probably represents the acquisition of a divergent neuroendocrine phenotype. The significance of the latter is unclear, although it may constitute a marker for malignancy.

Background: The human jaw-closing and jaw-opening muscles produce forces leading to the development of three-dimensional bite and chewing forces and to three-dimensional movements of the jaw. The length of the sarcomeres is a major determinant for both force and velocity, and the maximal work, force, and shortening range each muscle is capable of producing are proportional to the architectural parameter volume, physiological cross-sectional area, and fiber length, respectively. In addition, the mechanical role the muscles play is strongly related to their three-dimensional position and orientation in the muscle-bone-joint system. The objective of this study was to compare relevant architectural characteristics for the jaw-closing and jaw-opening muscles and to provide a set of data that can be used in biomechanical modeling of the masticatory system.

Methods: In eight cadavers, sarcomere lengths, muscle masses, fiber lengths, pennation angles, and physiological cross-sectional areas were determined for the following muscles: superficial and deep masseter, anterior and posterior temporalis, anterior and posterior medial pterygoid, inferior and superior lateral pterygoid, posterior and anterior digastric, geniohyoid, posterior and anterior mylohyoid, and stylohyoid. To determine the spatial position of their action lines, the three-dimensional coordinates of the attachment sites were registered.

Results: Compared with the jaw openers, the jaw closers were characterized by shorter sarcomere lengths at the closed jaw, larger masses of contractile and tendinous tissue, larger physiological cross-sectional areas, larger pennation angles, shorter fiber lengths, shorter moment arms, and lower fiber-length-to-muscle-length ratios. In addition, architectural features differed across the muscles of the same functional group. Sarcomere length did not differ significantly among the regions of the same muscle. In contrast, in some muscles, significant intramuscular differences were found with respect to, e.g., physiological cross-sectional area, fiber length, pennation angle, and moment arm length.

Conclusions: The results suggest that the jaw-closing muscles have architectural features that suit them for force production. Conversely, the jaw-opening muscles are better designed to produce velocity and displacement.

Background: The increase in the distal air spaces which takes place with age is the only sufficiently documented datum for differentiation between the senile and the adult lung. There are other pulmonary components which may be modified as the lung enters the phase of old age, but they have not as yet been sufficiently studied. The purpose of this study is to make a morphometric comparison between the wall thickness of the distal air spaces, elastic fiber, and collagen in lungs of rats of between 5 and 18 months of age.

Methods: The left lung of each rat was histologically processed for light microscopy. The sections were contrasted using methylene blue, resorcinfuchsin and Sirius red. Systematic randomized sampling was used for the selection of the histological fields studied. Morphometric variables were studied, and were systematized into three groups, namely: variables related with the alveolar architecture, variables which quantify elastic fiber, and those which quantify collagen.

Results: The old animals exhibited significant differences (p < 0.05) in the following variables: 1. In relation with the alveolar architecture, mean linear intercept, alveolar chord and wall thickness increased, whereas internal alveolar perimeter and tissue density decreased. 2. The relation elastic fiber density/lung tissue density increased. None of the variables which quantify collagen displayed significant differences.

Conclusion: Our data lead us to consider that the lungs of the old animals displayed, not only an enlargement of the distal air spaces, but also a thickening of the alveolar wall and an increase in elastic fiber in relation to the rest of the lung tissue.

Background: The bursa of Fabricius provided the microenvironment for B-cell differentiation. Continuous contact between lymphoid cells and antigen in the bursa further suggested that antigenic material has an important influence on the maintenance and development of B cells in the bursa. In addition, a dendritic cell, the bursal secretory dendritic cell (BSDC), has been identified in the medulla. The hypothesis that, in the bursal follicles, the contact between the lymphoid cells and the antigen may be mediated by dendritic cells prompted us to identify a bursal dendritic cell that becomes activated after contact with the antigen.

Methods: A polyclonal antiserum to S-100 protein was used to identify bursal dendritic cells because S-100 protein, a calcium-binding protein, has been shown to be a marker for the identification of chicken dendritic cells following recent contact with antigen.

Results: At every age investigated, S-100-positive cells showed a location and shape identical to those described for BSDCs. Positive cells were found within and under the follicle-associated epithelial cells (FAE), indicating that these cells were strategically placed where they would encounter the antigen. In addition, positive cells were found arranged along the corticomedullary junction, which is a regenerative zone for the BSDC. After 10 weeks of age, the number of positive cells dramatically decreased, suggesting that the endocytic activity of the FAE may become impaired as the bursa regresses.

Conclusions: The polyclonal antiserum to S-100 protein identified the BSDCs in the bursal follicles. Positive cells may be BSDCs that have undergone a functional activation after contact with the antigen. These cells may have a role as antigen-presenting cells in the bursal follicles. Hence, these cells may be involved in the events that lead to B-cell differentiation.

Background: Osteoclast colony stimulating factor (O-CSF) is an osteoclast-specific growth factor that stimulates the clonal growth of primitive osteoclast progenitors from bone marrow cells in culture. To characterize the morphology of immature osteoclasts (preosteoclasts) arising from complex hematopoietic tissues, progenies of O-CSF-responsive progenitors were cocultured with devitalized calvariae, and their cytochemical and ultrastructural features were examined.

Methods: Murine bone marrow cells were cultured in semisolid medium for 14 days in the presence of O-CSF. Colonies derived from osteoclast progenitors were then cocultured with devitalized mouse calvariae for 5 days. Cells attached to the calvariae were stained for tartrate resistant acid phosphatase (TRAPase), and the ultrastructure of these cells was examined by transmission electron microscopy. Bone marrow cells stimulated by macrophage colony stimulating factor (M-CSF) were similarly studied as a control.

Results: O-CSF-induced preosteoclasts stained strongly positive for TRAPase when cocultured with calvariae. These cells showed single nuclei, and their cytoplasm contained numerous mitochondria, vacuoles, granules, and coated vesicles. The ruffled cell border consisted of short, blunt, fingerlike projections. The adjacent clear zone contained abundant microtubules, microfilaments, and long narrow channels. M-CSF-induced macrophages were TRAPase negative, with no ruffled borders or clear zones.

Conclusions: All the characteristic features of active osteoclasts were observed in the cells derived from O-CSF-responsive bone marrow progenitors except that these cells were mononucleated, and their ruffled borders were not fully convoluted, indicative of their immature nature. This study documents for the first time the ultrastructural characteristics of preosteoclasts derived from cultured bone marrow progenitors in early stages of development.