Helen Michaloudi

The Noradrenergic Innervation of Identified Hypothalamic Magnocellular Somata and its Contribution to Lactation-Induced Synaptic Plasticity

Despite several studies showing that the rat supraoptic (SON) and paraventricular (PVN) nuclei are innervated by noradrenergic afferents, the respective contribution of these inputs to the oxytocinergic and vasopressinergic neuronal populations remains to be clearly defined. In the present study, we used the unbiased disector method to estimate the numerical density of noradrenergic varicosities on identified oxytocinergic and vasopressinergic somata in the rat SON and PVN. The analysis was carried out on semithin (1 μm) plastic sections cut from vibratome slices (50 μm) of the SON and PVN which had been double‐labelled for noradrenaline (NA) and oxytocin‐ or vasopressin‐related neurophysin. These preparations displayed many noradrenergic varicosities which electron microscopy showed to represent, in the main, synaptic boutons. Our quantitative analysis revealed that noradrenergic varicosities contacted oxytocinergic and vasopressinergic somata to a similar extent in male and female rats, under basal conditions of hormone secretion. The incidence of these axo‐somatic contacts was similar in the SON and PVN. In contrast, in lactating rats, in which oxytocin secretion is enhanced, there was a significant increase in the density of noradrenergic varicosities apposed to oxytocinergic somata, in both nuclei. Our observations indicate that, in male and female rats under normal conditions, noradrenergic afferents innervate each type of neurosecretory somata, in both magnocellular nuclei, in a similar fashion. They reveal, moreover, that noradrenergic afferents participate in lactation‐induced structural plasticity of synapses impinging on oxytocinergic somata.

Vascular network of the rat hippocampus is not homogeneous along the septotemporal axis.

Qualitative and quantitative image analysis of hippocampal vascular bed, after transcardial perfusion of India ink, reveals significant differences among hippocampal subfields and along the septotemporal axis of the rat hippocampus. Ventral hippocampus exhibits significantly higher levels of vascularization compared to dorsal hippocampus, which, however, is characterized by significantly higher capillary density. These results may explain the selective ischemia vulnerability of hippocampus along its septotemporal axis.

Mast cells in the sheep, hedgehog and rat forebrain.

The study was designed to reveal the distribution of various mast cell types in the forebrain of the adult sheep, hedgehog and rat. Based on their histochemical and immunocytochemical characteristics, mast cells were categorised as (1) connective tissue‐type mast cells, staining metachromatically purple with the toluidine blue method, or pale red with the Alcian blue/safranin method, (2) mucosal‐type or immature mast cells staining blue with the Alcian blue/safranin method and (3) serotonin immunopositive mast cells. All 3 types of brain mast cells in all species studied were located in both white and grey matter, often associated with intraparenchymal blood vessels. Their distribution pattern exhibited interspecies differences, while their number varied considerably not only between species but also between individuals of each species. A distributional left‐right asymmetry, with more cells present on the left side, was observed in all species studied but it was most prominent in the sheep brain. In the sheep, mast cells were abundantly distributed in forebrain areas, while in the hedgehog and the rat forebrain, mast cells were less widely distributed and were relatively or substantially fewer in number respectively. A limited number of brain mast cells, in all 3 species, but primarily in the rat, were found to react both immunocytochemically to 5‐HT antibody and histochemically with Alcian blue/safranin staining.

Expression of the prion protein in the rat forebrain - an immunohistochemical study

The cellular prion protein (PrP(C)) is crucial for the development of transmissible spongiform encephalopathies (TSEs), where the pathogenic scrapie isoform (PrP(Sc)) of the same protein, is considered to be the principal or sole infectious agent. Here, we report findings on PrP(C) expression in the rat forebrain, using immunohistochemical techniques on free floating sections of 60 microm thickness. Along with neurons and astrocytes in the gray matter, PrP(c) was detected for the first time in glial cells of the white matter and in cells of circumventricular organs. PrP(C) positive cellular processes were also found to be closely associated with intraparenchymal blood vessels, often in the form of end feet. Interestingly, PrP(C) expression was observed in areas where PrP(Sc) deposition in late stages of infection has been earlier reported in the rat and other species.

Acute inflammation alters adult hippocampal neurogenesis in a multiple sclerosis mouse model

Neural precursor cells (NPCs) located in the subgranular zone (SGZ) of the dentate gyrus (DG) give rise to thousands of new cells every day, mainly hippocampal neurons, which are integrated into existing neuronal circuits. Aging and chronic degenerative disorders have been shown to impair hippocampal neurogenesis, but the consequence of inflammation is somewhat controversial. The present study demonstrates that the inflammatory environment prevailing in the brain of experimental autoimmune encephalomyelitis (EAE) mice enhances the proliferation of NPCs in SGZ of the dorsal DG and alters the proportion between radial glial cells and newborn neuroblasts. The injection protocol of the cell cycle marker bromodeoxyuridine and the immunohistochemical techniques that were employed revealed that the proliferation of NPCs is increased approximately twofold in the SGZ of the dorsal DG of EAE mice, at the acute phase of the disease. However, although EAE animals exhibited significant higher percentage of newborn radial‐glia‐like NPCs, the mean percentage of newborn neuroblasts rather was decreased, indicating that the robust NPCs proliferation is not followed by a proportional production of newborn neurons. Significant positive correlations were detected between the number of proliferating cells in the SGZ and the clinical score or degree of brain inflammation of diseased animals. Finally, enhanced neuroproliferation in the acute phase of EAE was not found to trigger compensatory apoptotic mechanisms. The possible causes of altered neurogenesis observed in this study emphasize the need to understand more precisely the mechanisms regulating adult neurogenesis under both normal and pathological conditions.

Mast cells populations fluctuate along the spinal dura mater of the developing rat

The present study reveals developmental changes in the number, the phenotype and the distribution pattern of mast cells (MCs) along the cervical, the thoracic and the lumbar parts of the spinal dura mater. Postnatal infiltration of spinal dura by MCs does not appear to follow a sequential developmental pattern and meningeal MCs are unevenly distributed along the various parts of the examined dura. At each spinal level, areas most densely populated by MCs are the dorsal dura and the dural sleeves of the dorsal (sensory) spinal roots The developmental time course of the total MCs number is characterized by significant fluctuations in all three parts examined, with notable increases at P1, P4, P21 and P60 (peak value) for the cervical part, at P1 (peak value), P7 and P21 for the thoracic part and at P1, P7 (peak value) and P30 for the lumbar part. At P180, MCs number declines to 56%, 33% and 13% of the peak values for the cervical, the thoracic and the lumbar part, respectively. However, a different developmental pattern is followed by each subpopulation of MCs identified on the basis of their staining characteristics, namely connective tissue type mast cells (CTMCs), mucosal type or cells with characteristics of immature mast cells (MTMCs) and mixed type MCs, in each part examined. The findings may be of importance in elucidating physiological and pathological processes in the dura mater and the vertebral column.

Parallel development of blood vessels and mast cells in the lateral geniculate nuclei

The present study examined quantitatively developmental changes of the vasculature in the dorsal (dLGN) and the ventral (vLGN) lateral geniculate nuclei together with concomitant changes in the number of mast cells (MCs), known for their role in angiogenesis. Vascular network, marked after transcardial perfusion of India ink, and MCs detected with conventional histochemical techniques were examined at postnatal days (P) 1, 8, 14, 21, 31, 90 and 300 of Wistar rats. Quantitative analysis by means of an image analysis system showed age-dependent changes in both vascular parameters [vascular area and relative frequency (%) of capillaries and medium- and large-diameter vessels] and mast cells number in the developing dLGN and vLGN. Despite quantitative differences in the vascularization and MC infiltration between the two nuclei at some age points, MC number, vascular area and the percentage frequency of capillaries exhibited similar developmental time courses, especially up to the end of the first postnatal month. Both MC number and the capillary frequency reached maximal levels at P31 and declined thereafter, following a massive or a partial, respectively, decrease up to P300.

Developmental changes of mast cell populations in the cerebral meninges of the rat

It is known that both the dura and the pia mater attract and support the differentiation of mast cells. The present study shows that unevenly distributed mast cells in the cerebral meninges of the rat can be found in perivascular sites and vessel ramification points, but can also be unrelated to the meningeal vasculature. It also documents changes in the number, localization and staining preferences of the mast cells in the two meninges of the developing and mature rat brain. Quantitative examination of all types of histochemically differentiated meningeal mast cells reveals no major (although some exist) differences between right and left side subpopulations, but strongly suggests a different origin and fate of the dural and the pial mast cells. The number of dural mast cells, already high from postnatal day 0, although declining from postnatal day 21 onwards, remains conspicuous up to postnatal day 180. In contrast, pial mast cells are comparatively very few in the first day of the postnatal life, and despite a transient significant increase in the following two weeks, they reach almost zero levels from postnatal day 21.

Developmental changes in the vascular network of the rat visual areas 17, 18 and 18a

The present study examines quantitatively the areal and the laminar fluctuations of the vascular network in the visual areas 17, 18 and 18a of the rat cerebral cortex, from postnatal day (P) 1 to P60. For this purpose, the detailed vascular networks of the visual areas, marked after transcardial perfusion of India ink, are analyzed with the use of an image analysis system in order to measure the total vascular density (VD) and the relative density of capillaries (CD), of medium (MD)- and large (LD)-sized vessels in combination with changes in the mean diameter of all three types of vessels. Comparative quantitative microscopy showed that both VD and CD do not exhibit significant interareal differences in the adult rat brain. However, while VD reaches adult values much earlier in area 18a (P21) than in areas 17 and 18 (P60), CD obtains adult values at P31 in areas 17 and 18a, but later (P60) in area 18. Maturation process of laminar VD, CD, MD and LD was not found to follow a simple (i.e. inside-out or mediolateral) sequence, and, in each cortical area, laminar fluctuations of vessels density revealed a complicated developmental pattern, which might be attributed to their changing structural and functional status. Developmental changes in the diameter of capillaries, examined in conjunction with concomitant changes of vascular and capillary density in each area, suggest the existence of angiogenesis in all three visual areas during the third postnatal week of age.

Apoptosis in the visual system of normal and dark-reared rats

Apoptotic cell death, known to play a role in the developing brain, was immunocytochemically mapped in cortical and subcortical visual centers of Wistar rats developing under normal lighting conditions or in complete darkness. Apoptosis was relatively pronounced and exhibited the same developmental pattern in all visual centers examined. Visual deprivation was found to have negligible effects on apoptosis in subcortical areas, or to amplify it temporarily in the cerebral cortex.