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Mast cells in the amphibian brain during development
This is the first descriptive study of ontogenesis and anatomical distribution of mast cells in the developing brain of three different amphibian species. In the toad and the green frog, mast cells are preferentially located in: (i) the meningeal lining (pia mater), (ii) the choroid plexuses, both anterior and posterior, and (iii) the neuropil, in close association with the epithelial cell lining of blood vessels. It is only in the perennially aquatic African clawed frog that mast cells never appear inside brain ventricles and within the neuropil. Mast cells first become identifiable in brain of different species in different stages of development. While there are differences in the number of mast cells in different species at different stages of development, the number nearly doubles in all three species during the transition from pro-metamorphic stage of larval development to the peak of metamorphic climax. Furthermore, the number of mast cells is comparatively higher in the toad and remarkably lower in the fully aquatic Xenopus laevis, in which species the first appearance of identifiable mast cells during larval development occurs much later than in equivalent stages of development of the toad and the green frog. The secretory nature of mast cells can be assumed by the presence of cytoplasmic granules, which may show species-specific texture. Further experimental analyses are required to unveil the usefulness of mast cells in the amphibian brain. © 2010 The Authors. Journal compilation © 2010 Anatomical Society of Great Britain and Ireland
Occurrence of D-aspartate in the Harderian gland of Podarcis s. sicula and its effect on gland secretion
High concentrations of free D-aspartate (D-Asp), an amino acid well known for its neuroexcitatory activity, are endogeneously present in the Harderian gland (HG) of the lizard Podarcis s. sicula. This orbital gland consists of two different parts: the medial part, which is prevalently a mucous acinar gland, and the lateral part, which is a serous tubulo-acinar gland. To determine the physiological effect of D-Asp on exocrine secretion in HG, D-Asp (2.0 μmol/g b.w.) was injected intraperitoneally into lizards. We found that highest accumulations of exogenous D-Asp in HGs occurred 15 hr after the injection. Specifically, exogenous D-Asp prevalently stimulated serous secretion from the lateral portion of the gland, where immunohistochemical analysis revealed a major accumulation. Similarly, in the medial part of the gland, highly sulfated mucosubstances were observed after D-Asp injection. Further, in both parts of the HG, the electron microscope revealed euchromatic nuclei, a prominent rough endoplasmic reticulum, as well as numerous secretory granules within the acinar cells. Thus, following D-Asp injection, a 60% increase in HG total protein was detected. In addition, exogenous D-Asp induced changes in the electrophoretic pattern of HG. In conclusion, although further investigations are still needed to clarify the molecular pathway induced by D-Asp in exocrine secretion, this study does indicate that free D-Asp plays a significant role in the secretory activity of this gland. © 2006 Wiley-Liss, Inc
Induced Maturation of Frog Mast Cells by Nerve Growth Factor during Ontogenesis
The effect of nerve growth factor (NGF) on ontogenesis of frog mast cells was investigated in vivo by histochemical, morphometric, and ultrastructural analysis. Three groups of tadpoles at various stages of development were used. In the first group, the larvae received i.p. injections of 1 ng NGF/g; the second group received 10 ng NGF/g, while the control group received only the vehicle. The first recognizable mast cells arose symmetrically in the tongue at stage 26 of Witschi's standard table. At stages 26 and 29, the mast cell number in the NGF-injected tadpoles was significantly higher than the control group. From stage 29 onward, the mast cell number rapidly increased in all groups. No significant differences in mast cell number were observed between the control group and the NGF-injected groups at stages 31 and 33, Electron microscopy revealed that at metamorphic climax (stage 33), the mast cells in the NGF-treated groups were more mature than those in the control group. Therefore, nerve growth factor at early stages of tadpole development is likely to induce differentiation of mast cell precursors, while at later stages it is likely to induce maturation of immature mast cells. The close anatomical association between mast cells and perineurium, observed during nerve development, is intriguing. Already in the early stages of nerve development, the mast cells form a network around Schwann cell-axon complexes, together with the perineurial cells. At climax, the mast cells are located between the perineurial layers, suggesting that they may play a role in the tissue-nerve barrier of the perineurium. Nerve growth factor also seems to induce perineurial cell maturation. © 2003 Wiley-Liss, Inc
Morphological and biochemical changes in the Harderian gland of hypothyroid rats
The secretory activity of the Harderian gland (HG) is influenced by both exogenous (such as light and temperature) and endogenous (such as prolactin, thyroid hormones and steroid hormones) factors, which vary among species. In the present study, the effects of hypothyroidism on the rat HG were examined at morphological and biochemical levels. The decrease in cytoplasmic lipoproteic vacuoles and the increase in mucosubstance secretion in the acinar lumina were the most notable histological effects elicited by hypothyroidism. The release of all granules with nuclei and cellular debris suggested the occurrence of holocrine secretion. Electron microscopy revealed in the glandular cells of hypothyroid rat an increased condensation of chromatin in the nuclei, mitochondria with decreased cristae and vacuolisation, decreased glycogen granules, autophagic vacuoles, and lipofuscins in the cytoplasm. TUNEL reaction indicated DNA fragmentation in hypothyroid HG, indicative of an underlying apoptotic process. Translocation of cytochrome c from mitochondria to cytosol strongly supported this hypothesis. In conclusion, these findings indicate that thyroid hormones play a pivotal role in preserving the structural integrity of the rat HG and, hence, its secretory activity
Immunocytochemical evidence of the mast cell-fibronectin interaction in toad brain (Bufo bufo) during development
Mast cells in Wallerian degeneration: Morphologic and ultrastructural changes
The morphologic and ultrastructural changes of mast cells were followed in degenerating distal and regenerating proximal stumps of frog brachial nerve during Wallerian degeneration. Quantitative analysis included determination of both number and size of mast cells. The mast cell response to injury consisted of an early and a late phase. In the early phase, there was an increase in mast cell numbers in the proximal site of the lesion and a release of Alcian blue material consistent with mediator release. This phase of mast cell activation may be related, through the secretion of biogenic agents such as heparin and histamine, to the increase of endoneurial vessel size and vascular permeability, providing access for macrophages and mast cell precursors. The later phase, which peaked at 40 days after transection in the degenerating distal stump, consisted in the degranulation of the mast cells. These mast cells, closely associated with macrophages and degenerating Schwann cells, released secretory granules into the endoneurial microenvironment. These degranulating mast cells, through the released acid hydrolases, may contribute along with macrophages and Schwann cells, to the degradation of myelin debris. At the same time, mast cells appeared filled with granular content in the regenerating proximal segment. Therefore, we suggest that mast cells in peripheral nerves may play an important role in nerve degenerating and regenerating mechanisms through the secretion of diffusible molecules. © 2002 Wiley-Liss, Inc
Morphological changes in frog mast cells induced by nerve stimulation in vivo
We investigated at both histochemical and ultrastructural levels the effects of unilateral electrical stimulation in vivo of the frog hypoglossal nerve on the mast cells (MCs) within the nerve fascicles and among the axon terminals. The right ventral root of the hypoglossal nerve in different experiments was stimulated respectively for 1, 3, 5, 10 min with over-threshold stimuli (10 Hz; 2 ms duration). The stimulations at 3, 5 and 10 min caused a progressive degranulation and histochemical and ultrastructural changes of the MCs at the stimulated side. The morphological changes consisted of the loss of Alcian Blue secretory content and of a progressive release of safranin + secretory granules, depending upon duration of stimulation. The ultrastructural study showed that granules are discharged whole into the microenvironment or may release their content through exocytosis. A functional relationship between nerve and MCs is also suggested by the close anatomical association between MCs and pre-terminal axons observed following 10 min of hypoglossal stimulation. No changes in MC morphology occurred after 1 min of electrical stimulation. The results suggest that active cholinergic fibres can modulate MC secretion. © 2001 Elsevier Science Ireland Ltd. All rights reserved
Androgen and estrogen receptors expression in the rat exorbital lacrimal gland in relation to "harderianization"
The rat exorbital lacrimal glands (ELG) are particularly interesting for their biochemical and morphological sexual differences. Our histochemical and ultrastuctural observations confirm a phenomenon termed " harderianization" that occurs in the ELG of males and females at three months of age. The "harderianization" consists of the appearance of lipid foci in the ELG; this effect increases at six months of age only in the male glands, while it is not detectable in those of females. Histochemical tests for mucosubstances and proteins evidenced that while the secretory granules of male ELG are prevalently composed of sulphate substances, those of the female are composed of acid substances, and only a few cells positive to proteins were seen in the acinar epithelium of the glands. Moreover, we demonstrated by RT-PCR the presence of androgen and estrogen receptors in the rat ELG of both sexes. Androgen receptor transcript is always present in male and female ELG while the expression of estrogen receptor is not more detectable in the ELG of males at six months of life. In conclusion, our results suggest that estradiol may prevent the further lipid degeneration of the female ELG at six months of life. In addition, the disappearance of both the "harderian lipid" foci in the female gland and of estrogen receptor in the male gland indicates a probable involvement of estrogens in the phenomenon of "harderianization." © 2004 Wiley-Liss, Inc
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