The distribution of the projection from the hippocampal formation to the nucleus accumbens in the rat: an anterograde- and retrograde-horseradish peroxidase study

We have investigated the distribution and organization of the projection from the hippocampal formation to the nucleus accumbens in the rat. In the first experiments, horseradish peroxidase was injected into the fimbria fornicis. This procedure resulted in massive anterograde labelling of fornix fibers, and enabled the hippocampo--accumbens projection to be charted in detail. Labelled fornix fibers are distributed to the entire length of the nucleus accumbens and do not spread lateral to the anterior limb of the anterior commissure, that is, the projection is limited to the medial nucleus accumbens. Terminal labelling is particularly dense in a caudal dorsomedial sector of accumbens immediately adjacent to the septum. In the second part of the study, horseradish peroxidase was microelectrophoretically injected into the nucleus accumbens, in order to confirm the findings from the anterograde experiments. When the deposit was localized within the fornix distribution field, as indicated by the first experiment, retrogradely-labelled cells were observed in the subiculum, prosubiculum, and to a lesser extent, in hippocampal field CAI. When the deposit was located outside of the fornix distribution field, no hippocampal labelling was noted. The topography of the projection, suggested by the retrograde labelling experiments, is discussed. The findings confirm earlier reports of such a projection from the hippocampal formation to the nucleus accumbens, and with the use of a relatively novel anterograde-horseradish peroxidase technique, provide a picture of fiber labelling representing the entire field of origin of the fornix system. The striatal projection field of this pathway is discussed in relation to other striatal afferents, with particular emphasis on limbic afferentation of the striatum. Finally, the so-called 'hippocampal district' of striatum is discussed with respect to its neurotransmitter composition, as well as its functional importance regarding limbic system influence on central motor mechanisms.     

The amygdalostriatal projection in the rat—an anatomical study by anterograde and retrograde tracing methods

Tritiated leucine and proline injected into the amygdaloid complex was found to label a voluminous amygdalostriatal fiber system which is distributed to all parts of the striatum except an antero-dorsolateral striatal sector. The connection is established by way of the longitudinal association bundle as well as the stria terminalis, and includes a modest (10–15%), symmetrically distributed contralateral component conveyed by the anterior commissure. Both autoradiographic findings and subsequent observations in retrograde cell-labelling (horseradish peroxidase) material indicate that the amygdalostriatal projection originates mainly from the nucleus basalis lateralis amygdalae, in much lesser volume from the nucleus basalis medialis, and minimally from the nucleus lateralis amygdalae; no other contributing amygdaloid cell group could be identified.A comparison of the present findings with earlier reports indicates that the amygdalostriatal projection widely overlaps the striatal projections from the ventral tegmental area, the mesencephalic raphe nuclei and the prefrontal cortex. Like the amygdalostriatal projection, these striatal afferents largely or entirely avoid the antero-dorsolateral striatal quadrant, which thus appears to be the striatal region most sparsely innervated by afferents originating from structures within the circuitry of the limbic system. Findings in additional autoradiographic material identify this relatively non-limbic striatal quadrant as the main region of distribution of the corticostriatal projection from the sensorimotor cortex.     

Extra-hypothalamic afferent inputs to the supraoptic nucleus area of the rat as determined by retrograde and anterograde tracing techniques

To detect neuronal cell bodies whose axon projects to the hypothalamic supraoptic nucleus, small volumes (10-50 nl) of 30% horseradish peroxidase or 2% fast blue solutions were pressure-injected into the area of one supraoptic nucleus of rats. Both dorsal and ventral approaches to the nucleus were used. In animals where the injection site extended beyond the limits of the supraoptic nucleus, retrogradely labelled cell bodies were found in many areas of the brain, mainly in the septum, the nucleus of the diagonal band of Broca and ventral subiculum in the limbic system; the dorsal raphe nucleus, the locus coeruleus, the nucleus of the dorsal tegmentum, the dorsal parabrachial nucleus, the nucleus of the solitary tract and the catecholaminergic A1 region in the brain stem; in the subfornical organ and the organum vasculosum of the lamina terminalis, as well as in the median preoptic nucleus. In contrast, when the site of injection was apparently restricted to the supraoptic nucleus, labelling was only clearcut in the two circumventricular organs, the median preoptic nucleus, the nucleus of the solitary tract and the A1 region. Injections of wheat germ agglutinin coupled with horseradish peroxidase (60-80 nl of a 2.5% solution) made in the septum and in the ventral subiculum anterogradely labelled fibers coursing in an area immediately adjacent to the supraoptic nucleus but not within it. In contrast, labelling within the nucleus was found following anterograde transport of tracer deposited in the A1 region and in an area that includes the nucleus of the solitary tract. Neurones located in the perinuclear area were densely labelled by small injections into the supraoptic nucleus; they may represent a relay station for some afferent inputs to the supraoptic nucleus. These results suggest that the supraoptic nucleus is influenced by the same brain areas which project to its companion within the magnocellular system, the paraventricular nucleus.     

The regional distribution and cellular localization of iron in the rat brain

The regional distribution and cellular localization of iron throughout the rat brain was determined with iron histochemistry. Densitometry was used to measure the intensity of stain of 51 iron-concentrating sites. Among the areas of highest iron content are the circumventricular organs, islands of Calleja, globus pallidus, ventral pallidum, substantia nigra pars reticulata, interpeduncular nucleus, dentate nucleus, and interpositus nucleus. Iron occurs most commonly in oligodendrocytes and in the fibrous network of the neuropil, but is also found in the interstitial spaces of circumventricular organs and in the tanycytes of the organum vasculosum of the lamina terminalis, median eminence, and walls of the third ventricle. In diverse areas throughout the brain—among them, the islands of Calleja, dentate gyrus of the hippocampal formation, lateral septal nucleus, and central amygdala—iron is found in association with the perikarya and neuronal processes of nerve cells.The overlapping distribution patterns of iron and γ-aminobutyric acid, enkephalin, and luteinizing hormone-releasing hormone suggest that the distribution of iron is related to its association with the metabolism of one or more neurotransmitters or neuroactive compounds.     

The defects in development and apoptosis of cardiomyocytes in mice lacking the transcriptional factor Pax-8

Backgroud

Cardiac-specific deletion of ALK3 is lethal in mid-gestation with ventricular septum malformations (VSM). This study was designed to define the Pax-8's role in heart development and cardiomyocyte apoptosis.

Methods

Pathologic changes in the hearts of Pax-8 or ALK3 knockout and wild type control mice were determined by light and electron microscopy. Analysis of cardiomyocyte apoptosis was performed by TUNEL. The effect of Pax-8 gene deficiency on caspase-3 activity was examined after transfecting Pax-8 siRNA into cultured myoblast cell line.

Results

Mice with ALK3 or Pax-8 gene knockout but not wild type control animals showed the development of VSM. Increased cardiomyocyte apoptosis was found in homozygotes. Echocardiography showed that Pax-8 homozygote mice developed malfunction of the heart. Furthermore, the caspase-3 activity was significantly higher in the cells treated with Pax-8 siRNA as compared to those treated with negative control siRNA in H9C2 (2-1) cell line.

Conclusions

The Pax-8 gene may play a crucial role in heart development and regulating cardiocyte apoptosis. Knockout of Pax-8 may exert a similar effect on myocardial morphology and apoptosis as those seen in ALK3 knockouts. Furthermore, the ventricular septum malformations could be partially attributed to accelerated cardiomyocyte apoptosis.     

Effect of smoking on immunity in human chronic periodontitis

Aim

Evaluate the effects of smoking on dendritic cells (DCs), cytokines, clinical periodontal parameters, and number of teeth in samples of human chronic periodontitis (CP).

Material and methods

Gingival samples were obtained from 24 smokers and 21 non-smokers with CP. Periodontal examination was carried out. Immunohistochemical staining was performed to identify Factor XIIIa+ immature, CD1a+ immature, and CD83+ mature DCs. The inflammatory infiltrate was counted, and IL-2, IL-10, IL-4, IL-6, IFN-γ, TNF-α, and IL-17A were measured using the cytometric bead array (CBA). Inflammatory infiltrate, DCs, cytokines, classification of CP, clinical periodontal parameters, number of teeth, smoking habit in years (SH/years), and number of cigarettes smoked per day (C/day) were correlated and compared.

Results

CD83+ mature DCs decreased in the smokers group. Negative correlations could be observed between the number of C/day with levels of IL-17A and number of teeth. Correlations between smoking, periodontal disease status, and other cytokines were not observed.

Conclusions

Smoking decreases mature DCs in chronic periodontitis. Moreover, a dose-dependent relation can be observed between C/day and number of teeth and levels of IL17A observed. Smokers show a different modulation of the CP immune response.     

A study of the origin of brain stem projections to monkey spinal cord using the retrograde transport method.

We identified brain stem neurons projecting to cervical and lumbar levels of the spinal cord in young rhesus monkeys using the retrograde transport method. The somatotopic organizations of the red nucleus and lateral vestibular nucleus were clarified. In addition, the presence of bulbospinal neurons in the medial vestibular nucleus; the nucleus of the tractus solitarius; the medial and lateral reticular formations; the raphe nuclei magnus, obscurus, and pallidus; the hypothalamus; and the nuclei of the locus ceruleus and subceruleus was confirmed.     

Anatomical and physiological evidence for a cerebellar nucleo-cortical projection in the cat

Combined neuroanatomical and electrophysiological experiments were performed to test the hypothesis that axon collaterals of neurons in the cerebellar nuclei project to the cerebellar cortex in cats. The anatomical studies demonstrated that (a) following the injection of tritiated leucine into the deep cerebellar nuclei, labeled fibers could be traced into the granular layer of the cerebellar cortex, and (b) following the injection of horseradish peroxidase into the cerebellar cortex, retrogradely labeled horseradish peroxidase-positive neurons were identified in the deep nuclei. The electrophysiological experiments confirmed the anatomical findings. Neurons in the dentate and interposed nuclei, identified by their antidromic activation from the brachium conjunctivum, could also be activated antidromically from the cerebellar surface. Collision experiments demonstrated that projections from the deep cerebellar nuclei to the cerebellar cortex are in part collaterals of efferent neurons projecting through the brachium conjunctivum. Care was taken to ensure that all recordings were obtained from the region of cell somata in order to minimize the likelihood of recording from neuronal elements passing through the cerebellar nuclei. These combined neuroanatomical and electrophysiological studies provide strong evidence supporting the existence of a collateral system from cerebellar output neurons to the cerebellar cortex. The existence of this collateral system emphasizes that the cerebellar cortex and cerebellar nuclei may comprise a functional unit in which these collaterals may serve as a substrate for feedback control of the cerebellar cortex by the cerebellar output.     

Notch1 signaling,hippocampal neurogenesis and behavioral responses to chronic unpredicted mild stress in adult ischemic rats

Accumulating evidence indicates that the Notch signaling pathway fulfills important roles in ischemia-stimulated neurogenesis, which may be regarded as an etiological factor in post-stroke depression. Here we explored Notch₁ signaling, hippocampal neurogenesis and behavioral responses to chronic unpredicted mild stress (CUMS) in adult ischemic rats. Animals were treated with permanent middle cerebral artery occlusion followed by an 18 day CUMS procedure. Proliferating cells in the hippocampus and their cell fate were investigated on days 19 and 28 after ischemic surgery. Additionally, expression of the Notch₁ intracellular domain (NICD) and its downstream targets Hes1 and Hes5 was examined. A sucrose preference test and forced swim test were used to assess behavioral responses. CUMS produced depressive-like behaviors and decreased the number of proliferating cells on day 19 (both p < 0.001), accompanied by a decreased expression of both Hes1 and Hes5 in the hippocampus of ischemic animals (p < 0.001). On day 28, CUMS resulted in a decreased number of neurogenically-differentiating cells in the subgranular zone (p < 0.001) while permitting differentiation into astrocytes in the hilus (p < 0.05). Hes1 and Hes5 protein expression levels were increased. The expression of the NICD was significantly decreased at both time-points. CUMS led to expression changes in the Notch₁ signaling cascade in ischemic rats, most of which concerned hippocampal neurogenesis. This suggests that variation in Notch₁ activity and subsequent expression of its downstream targets, including Hes1 and Hes5, may, at least in part, contribute to modulation of ischemia-related hippocampal neurogenesis by CUMS.     

Regulation of enterocyte apoptosis by acyl-CoA synthetase 5 splicing

BACKGROUND AND AIMS: The constant renewal of enterocytes along the crypt-villus axis (CVA) of human small intestine is due to cell-inherent changes resulting in the apoptotic cell death of senescent enterocytes. The aim of the present study was to examine underlying molecular mechanisms of the cell death at the villus tip. METHODS: Characterization of human acyl-coenzyme A (CoA) synthetase 5 (ACSL5) was performed by cloning, recombinant protein expression, biochemical approaches, and several functional and in situ analyses. RESULTS: Our data show that different amounts of acyl-CoA synthetase 5-full length (ACSL5-fl) and a so far unknown splice variant lacking exon 20 (ACSL5-Delta 20) are found in human enterocytes. In contrast with the splice variant ACSL5-Delta 20, recombinant and purified ACSL5-fl protein is active at a highly alkaline pH. Over expression of ACSL5-fl protein is associated with a decrease of the anti-apoptotic FLIP protein in a ceramide-dependent manner and an increased cell-surface expression of the death receptor TRAIL-R1. Expression analyses revealed that the ACSL5-fl/ACSL5-Delta 20 ratio increases along the CVA, thereby sensitizing ACSL5-fl-dominated cells at the villus tip to the death ligand TRAIL, which is corroborated by functional studies with human small intestinal mucosal samples and an immortalized human small intestinal cell line. CONCLUSIONS: Our results suggest an ACSL5-dependent regulatory mechanism that contributes to the cellular renewal along the CVA in human small intestine. Deregulation of the ACSL5-fl/ACSL5-Delta 20 homeostasis in the maturation and shedding of cells along the CVA might also be of relevance for the development of intestinal neoplasia.