Ultrastructural response of arcuate nucleus neurons to fasting in aged rats.

The arcuate nucleus of the hypothalamus (ARH) is involved in the control of energy homeostasis. Leptin - an adipocyte derived hormone - is known to act on the hypothalamic nuclei and thus to control body weight by food intake reduction. Oxidative stress is believed to be implicated in leptin signalling. However, its relevance for leptin-induced signal transduction within ARH remains unclear. The goal of the study was to investigate the effect of fasting on morphological alterations of the neuronal endoplasmic reticulum/Golgi network as well as on the expression of leptin receptors in the arcuate nucleus of aged rats. Male Wistar rats, aged 24 months, were fasted for 96 hours. The control animals were fed ad libitum. Membranous whorls in the ARH neurons were visualized using the electron microscopy technique. Leptin receptors in the membranes of ARH neurons were determined immunohistochemically (IHC), and soluble leptin receptors in the plasma as well as plasma isoprostanes were quantified immunochemically (ELISA). An intense formation of membranous whorls was observed, directly associated with the cisternae of the rough endoplasmic reticulum, as well as lamellar bodies. Interestingly, the whorls were often localized near a well-developed Golgi complex. Moreover, some Golgi complexes displayed an early stage of whorl formation. Groups of residual lipofuscin granules were found in the immediate proximity of the whorls. An increased immunoreactivity with neuronal leptin receptors suggests that hypersensitive neurons may still effectively respond to the fasting serum levels of leptin, mediating ultrastructural transformation of ARH neurons during short-term fasting. Having observed a significant accumulation of lipofuscin granules and a marked increase of total 8-isoprostane serum level in the fasting rats, we hypothesize that signal transduction within the neurons of ARH is dependent on oxidative stress phenomena.

The effect of fasting on the ultrastructure of the hypothalamic arcuate nucleus in young rats.

In the present study, we described ultrastructural changes occurring in the neurons of the hypothalamic arcuate nucleus after food deprivation. Young male Wistar rats (5 months old, n = 12) were divided into three groups. The animals in Group I were used as control (normally fed), and the rats in Groups II and III were fasted for 48 hours and 96 hours, respectively. In both treated groups, fasting caused rearrangement of the rough endoplasmic reticulum forming lamellar bodies and membranous whorls. The lamellar bodies were rather short in the controls, whereas in the fasting animals they became longer and were sometimes participating in the formation of membranous whorls composed of the concentric layers of the smooth endoplasmic reticulum. The whorls were often placed in the vicinity of a very well developed Golgi complex. Some Golgi complexes displayed an early stage of whorl formation. Moreover, an increased serum level of 8-isoprostanes, being a reliable marker of total oxidative stress in the body, was observed in both fasting groups of rats as compared to the control.

Ultrastructural observations on the hypothalamic arcuate nuclei of aged rats in the fasting/refeeding model.

The arcuate nucleus of the hypothalamus (ARH) is involved in the control of energy homeostasis. This is the first study on the ultrastructural response of ARH neurons in aged rats after short-term fasting and subsequent refeeding. Male Wistar rats (24 weeks old) were fasted for 48 or 96 hours and were then refed for 24 hours. The controls were normally fed. The rats received water ad libitum. In both groups of fasting animals, we observed a rearrangement of the arcuate rough endoplasmic reticulum (RER) and Golgi complexes to form membranous whorls. Moreover, refeeding for 24 hours did not reverse this process. The RER was frequently found to be well organized into lamellar bodies composed of several cisternae. The membranous whorls were composed of concentric layers of endoplasmic reticulum and Golgi complexes. In addition, multiform lipofuscin granules were observed in close relationship with Golgi complexes and membranous whorls. Lipofuscin granules within the neurons of the arcuate nucleus are assumed to be a morphological manifestation of oxidative stress phenomena, which are presumably implicated in the formation of membranous whorls in both fasting and fasting/refed animals. This observation correlates with a significant increase in 8-isoprostane serum levels in the fasting and fasting/refed animals as compared to the fed control rats.

Changes in the morphology of the acinar cells of the rat pancreas in the oedematous and necrotic types of experimental acute pancreatitis.

Limited experimental models of the oedematous and necrotic types of acute pancreatitis provide some understanding of the pathophysiology of this disease. Wistar rats were treated with cerulein at 10 mg/kg of body weight or with L-arginine at 1.5 or 3 g/kg of body weight in order to induce the oedematous or necrotic type of acute pancreatitis. After the induction period we examined samples of pancreata with light and electron microscopes. Morphological examination showed profound changes in the histology of the pancreas and its acinar cells and subcellular structures, especially in the group of rats which received a higher dose of L-arginine, amounting to 3 g/kg body weight. These included parenchymal haemorrhage and widespread acinar cell necrotic changes. 4-OH-TEMPO successfully prevented morphological deterioration as well as amylase release, suggesting that the severity of the two types of disease strongly depends on the intensity of the oxidative stress. Our results lend support to the assumption that reactive oxygen species play an axial role in the pathogenesis of both types of acute pancreatitis.

Morphological changes of aortal smooth muscle cells exposed to calcitriol in culture.

INTRODUCTION: A number of observations suggest that vitamin D plays an important role in maintaining normal cardiovascular function, through its receptors in cardiac muscle or in aortal smooth muscle cells (SMCs). The purpose of this study was to determine the effect of vitamin D active metabolite--calcitriol [1,25(OH)2D3] on aortal SMCs. MATERIAL AND METHODS: The cells were isolated from the aortal media of newborn rats by enzymatic digestion and maintained for 6 weeks in primary culture. 1.2 microM of calcitriol was added to the medium every second day. RESULTS: Light microscopic investigations revealed that the control SMCs formed multilayer with characteristic 'hills and valleys'. Uniform cellular growth patterns were observed after calcitriol treatment. Ultrastructural studies indicated more rapid modulation of SMCs from the 'contractile' to the 'synthetic' phenotype following calcitriol treatment. Elastic fibers were more abundant in treated than in control cultures. The scanning electron microscope showed an increase of regular microvilli on the surface of SMCs. Morphometrical analysis of nuclei demonstrated phenotypic heterogeneity in populations of aortal SMCs. These results suggest that 1) calcitriol can induce changes in the phenotype and in the growth pattern of aortal SMCs, which may be associated with the onset or progression of the atherosclerosis process, 2) the basis of phenotypic changes is related to the stimulation of proliferative activity in these cells.

The amygdaloid body of the rabbit--a morphometric study using image analyser.

The amygdaloid body is a telencephalic structure belonging to the limbic system. The amygdaloid body consists of the two main nuclear groups: corticomedial and basolateral. The former-phylogenetically older group is composed of the central, medial, and cortical nuclei, while the latter, phylogenetically younger one, of the lateral, basolateral and basomedial ones. The results presented in our paper indicate differences in the structure and topography of the specific amygdaloid nuclei. Their subdivisions in the rabbit are not as evident as in the rat. Apart from structural differences, the cellular composition of specific nuclei does not differ distinctly. It can suggest that their intrinsic and extrinsic connections might be similar and the role and function of them is maintained (with few exceptions) through the phylogeny.

Ultrastructural organization of the visual zone in the claustrum of the cat.

Data obtained by using ultrastructural and morphometric approaches revealed three types of neurons in the cat visual claustrum. The most numerous were medium-sized and large ones. They formed 3/4 of the cell population. The ultrastructural properties of those cell types were largely similar. Their cell bodies were oval, round, fusiform or triangular and contained more or less indented nuclear envelope. The cytoplasm of those cells was characterized by a high concentration of subcellular organelles and particularly rough endoplasmic reticulum. The characteristic feature of those cells was a low nucleus/cell body area ratio (47 +/- 1% and 43 +/- 1%, respectively). The proximal dendrites of medium-sized cells were usually wide at the base, relatively short and tapering, whereas, those arising in the large cells were often thick and had a short tapering base. The neurons described above stained by Golgi impregnation method showed spines on their distal dendrites both under the light and electron microscopy. The retrograde axonal transport of HRP and WGA-HRP following injections into the visual cortex confirm that they are mainly projection cells which form the ascending limb of the claustrocortical loop. The third type of neurons formed a less numerous group of small cells which differed from the larger ones in various respects. They possessed the large nuclei with deeply indented nuclear envelope and comparatively a thim layer of cytoplasm poor in subcellular organells among which free ribosomes and mitochondria were common. The nucleus/cells body area ratio high (59 +/- 2%). In Golgi preparations their dendrites did not show spines. The dendrites originating from that type of neurons were thin, long and did not posses a wide tapering base. They are mainly claustral intrinsic neurons.

Relationship of corticoclaustral axon terminals to neurons of the claustrum in the cat.

A dark type degeneration of axon terminals was observed in the dorsocaudal part of the claustrum following lesions of the visual cortex. Most of the degenerating axon terminals had synapses mainly with dendritic spines (64.1%) and small dendrites (25.1%). Only 5.8% of degenerating axon terminals formed synaptic contacts with medium-sized dendrites, 1.0% with large dendrites and 0.5%--with other parts of the claustral neurons. 3.5% of degenerating axon terminals were observed to form a contact with more than one postsynaptic structure. Degenerating axon terminals displayed synaptic contacts exclusively of asymmetrical type with clearly identifiable postsynaptic densities which suggest corticoclaustral afferents to be excitatory. Our data strongly support the notion that there are direct synaptic contacts between the ascending arm of the corticoclaustral neuronal loop.

The distribution of axon terminals with flattened vesicles in the nuclei of the amygdaloid body of the cat.

The morphology of synapses in the amygdaloid nuclei was studied in 10 cats. On the basis of the percentage of axon terminals with flattened vesicles (F-type) nuclei were distinguished, in which these terminals are as sparsely distributed as in most areas of the central nervous system, from other nuclei in which they are abundant (about one-third to one-half of all synaptic boutons). The lateral, basal dorsal and basal ventral nuclei belong to the first, the medial and central nucleus and the anterior amygdaloid area--to the second group. The cortical nucleus, which generally has a small number of boutons of F-type has some parts seemingly belonging to the first, and others to the second group. In all amygdaloid nuclei axon terminals of F-type form symmetrical synaptic contacts. In nuclei with a low percentage of F-type terminals these boutons are predominantly small and synapse either with perikarya or with large dendrites. The amygdaloid nuclei having numerous F-type terminals contain not only small but also larger terminals with flattened vesicles. Both, the larger and smaller axon terminals form in these nuclei synaptic contacts with various parts of dendrites even with very small ones and with dendritic spines. The subdivision of amygdala into two parts, one with a low and another with a high number of F-type boutons would seem to support the hypothesis that amygdala may be subdivided physiologically into a dorsomedial--"excitatory" and basolateral--"inhibitory" portion.