Newly-formed neurons in the regenerating optic tectum of Triturus cristatus carnifex

A histochemical light and electron microscopy study of acetylcholinesterase (AChE) was carried out on the regenerating opic tectum of adult newt. A plug of optic tectum was removed and 15 days later [6-H3] thymidine was injected. Ninety days after the lesion the brain was removed, treated for histochemical AChE-detection and autoradiographic analysis. This double treatment showed the capacity of these adult amphibians to regenerate the nervous tissue through the proliferation of undifferentiated elements and their subsequent differentiation into neurons as is shown by the presence of cells both labelled by [6-H3] thymidine and by the AChE-reaction product.     

The effects of magnesium sulfate on blood-brain barrier disruption caused by intracarotid injection of hyperosmolar mannitol in rats

The study was performed to evaluate whether magnesium sulfate could alter the degree of disruption of the blood-brain barrier (BBB) caused by hyperosmotic mannitol. Wistar adult female rats were infused with 25% mannitol into the left internal carotid artery. Each animal received intraperitoneally a 300 mg/kg loading dose of magnesium sulfate, dissolved in 0.9% saline, followed by a further 100 mg/kg dose. In the other group, intracarotid infusion of magnesium sulfate was performed at a dose of 150 mg/kg 10 min before mannitol administration. Evans blue (EB) dye was used as a marker of BBB disruption. The measured serum glucose and magnesium levels increased after mannitol and/or magnesium administration when compared with their initial values before treatment (P < 0.01). Water content of the left hemisphere was significantly increased by hyperosmotic mannitol (P < 0.01). The increased water content in the mannitol-perfused hemisphere was significantly decreased by magnesium treatment (P < 0.05). The content of EB dye in the mannitol-perfused hemisphere markedly increased when compared with the right hemisphere of the same brain (P < 0.01). The EB dye content in the mannitol-perfused hemisphere following both intraperitoneal and intraarterial administration of magnesium decreased when compared with mannitol alone (P < 0.01). We conclude that although magnesium sulfate administration by both intracarotid arterial and intraperitoneal routes attenuates BBB disruption caused by hyperosmolar mannitol, particularly intraperitoneal route of magnesium sulfate administration may provide a useful strategy to limit the transient osmotic opening of the BBB.     

Morphogenesis of the adrenal gland of Triturus cristatus carnifex during larval development and metamorphosis

During the morphogenesis of the adrenal gland of Triturus cristatus, a cranio-caudal differentiation is observed together with a migration of the two cell types composing the adrenal gland: the steroidogenic cells and the chromaffin cells. During the cranio-caudal differentiation the two cell type gradually occupy an increasingly posterior position on the mesonephros until they are distributed, in the adult, along the whole kidney. The migration brings the cells from dorsal or dorso-lateral position, with respect to the venous vessels, to the ventral surface of the kidney, an arrangement typical of the adult.     

Absence of chiasmata from the heteromorphic region of chromosome I during spermatogenesis in Triturus cristatus carnifex

Analysis of squash preparations of spermatocytes from crested newts, Triturus cristatus carnifex, has shown that in most cells at least one large bivalent regularly fails to form chiasmata in one arm-pair. Feulgen microphotometry of diplotene and metaphase bivalents has shown that it is the largest bivalent in each cell which shows chiasma failure in one arm-pair. A C-banding technique which identifies chromosome I by virtue of a long, darkly stained region in its long arm, was used to confirm the absence of chiasmata from one arm-pair of the longest bivalent, and specifically from the darkly stained region. The achiasmate region which chromosome I exhibits during spermatogenesis, corresponds to the heteromorphic region of oocyte lampbrush bivalent I in which chiasmata never form. A possible correlation between the complete absence of crossing-over from the heteromorphic region and unusual cytological and molecular features which it exhibits, are discussed.     

Uptake of radioactive sulfate as a function of time by mast cells in Triturus cristatus carnifex (Laur.)

Summary Thirty two Triturus cristatus carnifex (Laur.) male individuals were injected intraperitoneally with a single dose of 150 c of carrier-free S³⁵ labelled sodium sulfate. The tissues were removed at different time intervals within a period of 3 hours and 30 days after the injection. Autoradiographs were prepared from mesentery whole mounts and histologie sections of the pelvic region. In mesentery whole mounts entire spider-shaped mast cells were studied. The earliest activity was always seen to arise in the perikaryon at 3 hours, where it persisted throughout the experiment while moving along the cellular processes progressively with time. The specific activity was assayed in the pelvic glands (bilateral organs of the reproductive system in males) and plotted against time. Since in these glands S³⁵O ₄ ^–– uptake in shown exclusively by mast cells the turnover time of 10–11 days therein recorded was taken to indicate the total metabolism of the sulfated compound stored in the mast cells. No direct evidence is as yet available to indicate that the metachromatic sulfated compound stored in Triturus mast cells is heparin.The present study was supported by a grant from the Consiglio Nazionale delle Ricerche-Impresa di Endocrinologia — Gruppo di Endocrinologia Comparata.     

Cell proliferation in vivo and in vitro in visceral organs of the adult newt, Triturus cristatus carnifex

The mitotic and labelling incidence of intestine, liver, spleen and pancreas cells of Triturus cristatus carnifex adults kept at 15°C, 20°C, 25°C and 30°C were examined. Intestine mitotic and labelling incidences were highest at 25°C and lowest at 30°C. There was no significant difference between 15°C and 20°C. No such relationship could be shown for liver, spleen or pancreas, which had very much lower mitotic and labelling incidences. In culture, intestine mitotic and labelling incidences fell significantly within the first four hours, and maintained these low levels for the next five days. In contrast, liver mitotic and labelling incidences rose for 9–11 days, and then began to fall, while pancreas mitotic and labelling incidences reached peak values at day 5, and were kept in good condition for up to 14 days.     

Proliferative response of the mesencephalic matrix areas in the reparation of the optic tectum of Triturus cristatus carnifex

The localization and proliferative response of optic tectum matrix cells has been studied in adult newt following an experimental lesion on an optic lobe. The results show that 15 days after the lesion the cells in division, autoradiographically labelled, are located in the periventricular layer. Thirty days after the lesion the labelled cells are also found in the innermost grey layers; at 90 days the injured optic tectum regains the cytoarchitecture characteristic of this centre, with labelled cells, whether in the external or in the internal pyriform layers. In all the stages the labelled cells are also found in the periventricular layers of the controlateral optic tectum, in the dorsal pallium and in the striatum. The quantitative data exhibit the existence of a direct relationship between the number of proliferating cells in the injured optic lobe and the extent of the lesion. These data show the possibility of active cellular proliferation for the reconstruction of the lesioned nervous area and for restoration of the characteristic histological structure.     

The organization of serotonin-immunoreactive neuronal systems in the brain of the crested newt,Triturus cristatus carnifex Laur.

Summary The distribution of serotonin (5-HT) immunoreactive structures has been investigated in the brain of the crested newt by means of indirect immunofluorescence, and unlabeled antibody peroxidase-antiperoxidase-complex (PAP) or biotin-avidin-system (BAS) techniques. In the newt, the bulk of the serotoninergic system extends from the raphe region of the medulla oblongata, through the isthmus, toward the mesencephalic tegmentum, and is characterized by pyriform neurons mainly located in a subependymal position, close to the midline. Also in the caudal hypothalamus, in addition to some 5-HT-positive adenohypophysial cells, many immunoreactive CSF-contacting neurons are found lining the paraventricular organ and the nucleus infundibularis dorsalis. A rich serotoninergic innervation was observed in the preoptic area and in the habenular complex. Concerning the telencephalon, immunopositive nerve fibers are encountered in the dorsal pallium, primordium hippocampi, striatum and olfactory bulbs. The general organization of serotoninergic systems in the newt brain exhibit close similarities to that described in higher vertebrates.