Differentiation of Cells in the Hypophysis and in Pancreatic Islets

Deparaffinized, 3-5μ, sections are brought to water, oxidized 3.5 min in an equal-parts mixture of 0.3% H₂SO₄ and 0.3% KMnO₄, and decolorized with 4% K₂S₂O₅. Nuclei are stained with Gomori's (1939) chromium-hematoxylin, and cell granules with Cason's (1950) mixture. The eosinophilic cells of the hypophysis and the alpha cells of pancreatic islets (of Langerhans) stain carmine red; basophilic and beta cells stain dark blue. Heidenhain's _susa is the most suitable fixative for hypophysis, Bouin's fluid for pancreas; but a satisfactory result is obtainable after formalin-sublimate or plain formalin. Besides studying the ratio of the cell types in the hypophysis or in pancreatic islets, it is possible to estimate the granule content of the cells. The method works on human autopsy material provided fixation of hypophysis occurs within 24 hr, and. pancreas, 12 hr post mortem, and it is suitable also for quite fresh organs.     

Cytology and cytochemistry of the hypophysis of the freshwater teleost Rhamdia hilarii (Val., 1840).

The characterization of cell types in the rostral pars distalis, proximal pars distalis and pars intermedia of the Bagre fish hypophysis during the reproductive cycles, as well as a cytochemical study of mucosubstances were undertaken. The cytophysiological relationship among the different acidophilic, basophilic and chromophobic cells with the several hypophyseal hormones was also considered.     

Differentiation of Cells in the Hypophysis and in Pancreatic Islets

Deparaffinized, 3-5μ, sections are brought to water, oxidized 3.5 min in an equal-parts mixture of 0.3% H₂SO₄ and 0.3% KMnO₄, and decolorized with 4% K₂S₂O₅. Nuclei are stained with Gomori's (1939) chromium-hematoxylin, and cell granules with Cason's (1950) mixture. The eosinophilic cells of the hypophysis and the alpha cells of pancreatic islets (of Langerhans) stain carmine red; basophilic and beta cells stain dark blue. Heidenhain's _susa is the most suitable fixative for hypophysis, Bouin's fluid for pancreas; but a satisfactory result is obtainable after formalin-sublimate or plain formalin. Besides studying the ratio of the cell types in the hypophysis or in pancreatic islets, it is possible to estimate the granule content of the cells. The method works on human autopsy material provided fixation of hypophysis occurs within 24 hr, and. pancreas, 12 hr post mortem, and it is suitable also for quite fresh organs.     

Global analysis of hematopoietic and vascular endothelial gene expression by tissue specific microarray profiling in zebrafish

In this study, we utilize fluorescent activated cell sorting (FACS) of cells from transgenic zebrafish coupled with microarray analysis to globally analyze expression of cell type specific genes. We find that it is possible to isolate cell populations from Tg(fli1:egfp)(y1) zebrafish embryos that are enriched in vascular, hematopoietic and pharyngeal arch cell types. Microarray analysis of GFP+ versus GFP- cells isolated from Tg(fli1:egfp)(y1) embryos identifies genes expressed in hematopoietic, vascular and pharyngeal arch tissue, consistent with the expression of the fli1:egfp transgene in these cell types. Comparison of expression profiles from GFP+ cells isolated from embryos at two different time points reveals that genes expressed in different fli1+ cell types display distinct temporal expression profiles. We also demonstrate the utility of this approach for gene discovery by identifying numerous previously uncharacterized genes that we find are expressed in fli1:egfp-positive cells, including new markers of blood, endothelial and pharyngeal arch cell types. In parallel, we have developed a database to allow easy access to both our microarray and in situ results. Our results demonstrate that this is a robust approach for identification of cell type specific genes as well as for global analysis of cell type specific gene expression in zebrafish embryos.     

Deconstructing the pharyngeal metamere

A prominent feature of all vertebrate embryos is the presence of a series of bulges on the lateral surface of the head, the pharyngeal arches. These structures constitute a metameric series, with each arch forming a similar set of derivatives. Significantly, the development of the pharyngeal arches is complex as it involves interactions between disparate embryonic cell types: ectoderm, endoderm, mesoderm and neural crest. It is becoming increasingly apparent that the development of the pharyngeal metamere revolves around the pharyngeal endoderm. The segmentation of this tissue is central to the generation of the arches. The pharyngeal endoderm also provides positional cues for the neural crest, and is involved in the induction of a number of components of the pharyngeal metamere. The segmentation of the pharyngeal endoderm has also been key to the evolution of pharyngeal metamerism. It is likely that endodermal segmentation is a deuterostome characteristic and that this basic pattern was sequentially modified and over time the more complex pharyngeal metamere of vertebrates emerged.     

Morphological aspects of the pharyngeal hypophysis in human embryos

In human embryos the hypophyseal sac (Rathke's pouch) originates at the roof of the mouth until stage 15 as a broad rim. As the mandibular arch and the maxillary swelling enhance the mesodermal masses in forming the early palatal shelves the rim is reduced to a cleft of about 0.2 mm in broadness in stage 17. From stage 18 up to stage 23 there is a prominent papilla in the midline of the mouth's roof which later on may become recanalized. The different SEM-aspects of the pharyngeal hypophysis are demonstrated.     

Endocrine cells share expression of N-CAM with neurones

The expression of the neural cell adhesion molecule, N-CAM, was examined in the anterior lobe of rat hypophysis by immunocytochemistry at light and electron microscope levels. In addition, N-CAM antigenic determinants present in adrenal medulla, anterior hypophysis and PC12 cells were compared by immunoblotting with those found in cerebellum. All secretory cells in the anterior hypophysis were found to be N-CAM positive on their surfaces, but not all of the three polypeptide determinants typical of cerebellum were present in the endocrine tissues or cell line tested. In addition, a new N-CAM determinant of 49 kDa not present in cerebellum was found in adrenal medulla and hypophysis, although it was absent from PC12 cells. The possible implications of these data are discussed.     

鮈亚科鱼类下咽骨和下咽齿的形态差异及其功能适应

采用双筒解剖镜观察和手工绘图的方法,研究鮈亚科鱼类39个代表种下咽骨和下咽齿的形态特征,分析鮈亚科鱼类下咽骨和下咽齿的形态差异及其功能适应。结果显示,鮈亚科鱼类的下咽骨可以分为粗壮型、中间型和狭长型3种形态类型;下咽齿可以分为圆锥型、臼齿型、粗壮侧扁型、侧扁型和极侧扁型5种形态类型。鮈亚科鱼类下咽骨和下咽齿间呈现出多种形态组合,并相互配合共同处理不同类型的食物。下咽骨和下咽齿的形态差异为鮈亚科鱼类摄食不同类型的食物、充分利用不同的生境资源在摄食器官多样性方面提供了保证。     

DiGeorge syndrome and pharyngeal apparatus development

DiGeorge syndrome is the most frequent microdeletion syndrome in humans, and is characterized by cardiovascular, thymic and parathyroid, and craniofacial anomalies. The underlying cause is disturbed formation of the pharyngeal apparatus, a transient structure present during vertebrate development that gives rise to endocrine glands, craniofacial tissue, and the cardiac outflow tract. The pharyngeal apparatus is composed of derivatives of ectoderm, endoderm, mesoderm and the neural crest. Thus, complex interactions between cell types from different origins have to be orchestrated in the correct spatiotemporal manner to establish proper formation of the pharyngeal system. The analysis of engineered mouse mutants developing a phenotype resembling DiGeorge syndrome has revealed genes and signalling pathways crucial for this process. Intriguingly, these mouse models reveal that interference with either of two distinct phases of pharyngeal apparatus development can contribute to the aetiology of DiGeorge syndrome.     

An Azocarmine stain for Differential cell Analysis of the rat Anterior Hypophysis

A method of staining is described which is especially designed to facilitate differentiation of the cell types of the rat anterior hypophysis. Fixation in Zenker-formol solution is recommended. Pre-staining of the nuclei by a short immersion in alum hematoxylin is followed by mordanting in anilin alcohol and a 45 minute period of staining in azocarmine solution at 60d`C. The counterstains, acid green and orange G, are dissolved in clove oil to avoid destaining of the azocarmine.     

Topographical distribution of the gonadotrophs,mammotrophs, somatotrophs and thyrotrophs in the pituitary gland of the baboon (Papio cynocephalus)

The identification and regionalization of four pituitary parenchymal cell types, gonadotrophs, mammotrophs, somatotrophs and thyrotrophs, were studied in the baboon (Papio cynocephalus) hypophysis using immunocytochemistry. The gonadotrophs were homogeneously distributed throughout the entire pars distalis. Both mammotrophs and somatotrophs predominate at the superior and inferior poles of the organ. The medial and anteromedial regions are populated by mammotrophs and thyrotrophs, while the lateral and posterior portions of the pars distalis contain large numbers of somatotrophs.     

Seasonal changes in the hypothalamo-hypophyseal-ovarian system in the catfish,Heteropneustes fossilis (Bloch)*

The annual reproductive cycle of the catfish, H. fossilis (Bloch) is divided into the preparatory period (February-April), the prespawning period (May-June), the spawning period (July-August) and the postspawning period (September-January). During the early postspawning period (September-November), the hypothalamo-hypophyseal-ovarian system shows a gradual regression. In January, the hypothalamic nuclei, the pars magnocellularis (PMC), the pars parvocellularis (PPC) of the nucleus preopticus (NPO), and the nucleus lateralis tuberis (NLT) show renewed activity, as shown by a significant increase in their nuclear diameters and an accumulation of neurosecretory material (NSM) in their cell bodies. The hypophysis and the ovary remain quiescent. During the preparatory period, all the hypothalamic neurons studied indicate decreased activity but simultaneously show an accumulation of NSM in their cell bodies. The number of granulated basophils in the proximal pars distalis (PPD) of the hypophysis remains low but ovarian weights increase, presumably due to the multiplication of oogonia. In the prespawning period, there is a marked accumulation of NSM in the cell bodies of the hypothalamic neurons and at the same time the number of granulated basophils in the PPD of the hypophysis dramatically increases with concomitant increase in vitellogenic activity in the ovary. During the spawning period, the hypothalamic neurons continue to store NSM in their cell bodies and simultaneously there is a tremendous increase in the number of granulated basophils in the PPD of the hypophysis and the ovary has a large proportion of yolky primary oocytes. Spawning is associated with a significant degranulation of the granulated basophils in the PPD of the hypophysis. The significance of the results is discussed in relation to the environmental and hormonal regulation of seasonal ovarian activity.     

Cellular structure and ultrastructure of the black coral Antipathes aperta: 2. The gastrodermis and its collar cells

The gastrodermis of the black coral Antipathes aperta is associated with eight distinct types of cells, including two types of microbasic b-mastigophores (nematocysts), spumous and vesicular mucus cells, and ganglion cells that are essentially the same as in the epidermis. Three additional types of cells are unique to the gastrodermis, and are readily distinguished from those of the epidermis by their electron-opaque inclusions. These include lipoidal cells, zymogen digestive cells, and an unusual type of epitheliomuscular collar cell. The pharyngeal region is characterized by the presence of electron-opaque nematocysts, a scattering of zymogen cells, and a large number of collar cells. The latter are distinguished in part by the presence of dense microfibrillar processes that surround the microvilli and extend into adjacent collars. This interconnection results in the formation of an extensive pharyngeal meshwork. These collar cells are additionally distinguished by the placement of the collar and flagellum adjacent to a flared cup of cytoplasm. This portion of the cell is capable of endocytosis of relatively large unicellular prey, and apparently is capable of forming digestive vesicles as well. The pharyngeal gastrodermis grades into simple lobate septal filaments toward the base of the coelenteron, where large, granular nematocysts all but replace the smaller electron-opaque types Collar cells are found here as well, but in fewer numbers compared to the zymogen cells. Ultrastructural results are compared with those of other coelenterates and discussed in terms of food and modes of nutrition.     

Auxin triggers transient local signaling for cell specification in Arabidopsis embryogenesis

The Arabidopsis embryonic root meristem is initiated by the specification of a single cell, the hypophysis. This event critically requires the antagonistic auxin response regulators MONOPTEROS and BODENLOS, but their mechanism of action is unknown. We show that these proteins interact and transiently act in a small subdomain of the proembryo adjacent to the future hypophysis. Here they promote transport of auxin, which then elicits a second response in the hypophysis itself. Our results suggest that hypophysis specification is not the direct result of a primary auxin response but rather depends on cell-to-cell signaling triggered by auxin in adjacent cells.     

Protofilament number in microtubules in cells of two parasitic nematodes

The parasitic nematodes, Ascaridia galli and Trichostrongylus colubriformis, were prepared for electron microscopy with fixatives containing tannic acid, which allowed their microtubule protofilament number to be examined. In contrast to many mammalian tissues, the nematodes did not contain microtubules with 13 protofilaments. Ascaridia galli contained microtubules with 11 protofilaments in all tissues examined, including nerve, intestinal, pharyngeal, and hypodermal cells. Trichostrongylus colubriformis contained nerve cells, known as microtubule cells, with bundles of larger microtubules (approximately 30 nm in diameter) with 14 protofilaments. The microtubules in these cells did not appear to be continuous for the entire length of the axon. Other cells examined in T. colubriformis, including nerve, intestinal and pharyngeal cells, contained two distinct types of microtubules, one with 11 protofilaments and an approximate diameter of 25 nm, and one with 12 protofilaments and an approximate diameter of 27 nm. All cell types examined contained both types of microtubules.     

The characteristics of the morphofunctional status of the adenohypophysis in lethally irradiated recipients after bone marrow transplantation]

The effect of lethal irradiation and transplantation of syngenic bone marrow in a dose of 1 x 10(7) cell/ml on the morphofunctional state of hypophysis at various stages of the posttransplantation period has been studied for 3 months using 100 linear male mice of a F1 (CBA x C57 B) line. The experiments conducted have shown that bone marrow transplantation reduces the intensity of the negative effect of irradiation on hypophysis and facilitates normalization of its histological structure. There was a correlation between changes in the number of secretory cells in the anterior lobe of the hypophysis and the level of corticosterone in irradiated and bone marrow-protected animals.     

Pharyngeal arch patterning in the absence of neural crest

Pharyngeal arches are a prominent and critical feature of the developing vertebrate head. They constitute a series of bulges within which musculature and skeletal elements form; importantly, these tissues derive from different embryonic cell types [1]. Numerous studies have emphasised the role of the cranial neural crest, from which the skeletal components derive, in patterning the pharyngeal arches [2-4]. It has never been clear, however, whether all arch patterning is completely dependent on this cell type. Here, we show that pharyngeal arch formation is not coupled to the process of crest migration and, furthermore, that pharyngeal arches form, are regionalized and have a sense of identity even in the absence of the neural crest. Thus, vertebrate head morphogenesis can now be seen to be a more complex process than was previously believed and must result from an integration of both neural-crest-dependent and -independent patterning mechanisms. Our results also reflect the fact that the evolutionary origin of pharyngeal segmentation predates that of the neural crest, which is an exclusively vertebrate characteristic.     

Hypophysis: a new scheme of ontogenetic development

It is usually accepted that hypophysis originates from two ecotodermal primordiums that give rise to its two main parts. The transformation of initial primordiums into definitive hypophysis is regarded as a result of mutual inductive influences between hypothalamus and Rathke's pouch. This scheme was remaining unchanged during many decades. Based upon the arguments that Spemann's organizer, prechordal mesoderm (PCM) and hypophysis are consecutive derivatives that replace each other during ontogeny, the author proposes that the two novel stages preceding the former starting point should be added to traditional scheme of pituitary development. Besides, according to new scheme the PCM (successor of Spemann's organizer) plays the main role in hypophysis development by active stimulation of morphogenesis in adjacent tissues. This leads to formation of Rathke's pouch and infundibulum, to their approaching each other, and to reprogrammin of Rathke's pouch ectoderm into hormonal cells of adenohypophysis. According to new hypothesis role of Rathke's pouch is accessorial and consists in provision of cell material.