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Background: Autophagy is a highly regulated process involving the bulk degradation of cytoplasmic macromolecules and organelles in mammalian cells via the lysosomal system. Dysregulation of autophagy is implicated in the pathogenesis of many neurodegenerative diseases and integrity of the autophagosomal - lysosomal network appears to be critical in the progression of aging. Our aim was to survey the expression of autophagy markers and Amyloid precursor protein (APP) in aged bovine brains. For our study, we collected samples from the brain of old (aged 11-20 years) and young (aged 1-5 years) Podolic dairy cows. Formalin-fixed and paraffin embedded sections were stained with routine and special staining techniques. Primary antibodies for APP and autophagy markers such as Beclin-1 and LC3 were used to perform immunofluorescence and Western blot analysis. Results: Histologically, the most consistent morphological finding was the age-related accumulation of intraneuronal lipofuscin. Furthermore, in aged bovine brains, immunofluorescence detected a strongly positive immunoreaction to APP and LC3. Beclin-1 immunoreaction was weak or absent. In young controls, the immunoreaction for Beclin-1 and LC3 was mild while the immunoreaction for APP was absent. Western blot analysis confirmed an increased APP expression and LC3-II/LC3-I ratio and a decreased expression of Beclin-1 in aged cows. Conclusions: These data suggest that, in aged bovine, autophagy is significantly impaired if compared to young animals and they confirm that intraneuronal APP deposition increases with age. [ABSTRACT FROM AUTHOR]

The aim of the study was to estimate developmental changes in the rabbit basolateral complex (BLC) by stereological and histochemical methods. Material consisted of 45 brains of New Zealand rabbits (aged from 2 to 180 days, P2 to P180) of both sexes, divided into nine groups. The following parameters were estimated: volume of the cerebral hemisphere; volume of the whole BLC and of particular BLC nuclei; neuronal density and total number of neurons in these nuclei. Developmental changes in acetylcholinesterase (AChE) activity in the BLC were also examined. The volume of the cerebral hemisphere increased until P30, whereas volumes of nuclei increased for longer – until P90. The density of neurons in all nuclei studied reached the level characteristic for an adult animal at about P30. The total number of neurons in the dorsolateral division of the lateral nucleus (Ldl) stabilized the earliest – between P30 and P60, whereas in the ventromedial division of the lateral nucleus (Lvm), basomedial (BM) and basolateral (BL) nuclei the number stabilized later – between P60 and P90. AChE activity appears minimal in the BLC on P2, reaches a maximum on P30 and then decreases to the level characteristic of an adult animal on P60. AChE activity was greater in BL than in other nuclei in all age groups. Reaching adult AChE activity 1 month earlier than the total number of neurons in the BLC may indicate a role of the cholinergic system in BLC maturation. [ABSTRACT FROM AUTHOR]

Calbindin-D28k (CB), parvalbumin (PV) and calretinin (CR) are calcium-binding proteins (CaBPs) considered to be markers for certain subpopulations of neurons in the central nervous system. The aim of this study was to describe the pattern of distribution of CB-, PV- and CR-immunoreactive elements in the rabbit corticomedial amygdaloid complex during the postnatal period. The time course of changes in CaBPs expression during maturation of the selected nuclei indicates their diversity. During the first month after birth, CaBPs expression stabilizes earliest in the anterior cortical and then in the medial nuclei. Later, during the second month of postnatal life, the posteromedial and posterolateral cortical nuclei maturate. The central nucleus requires a considerably longer time to reach maturity - about three months are needed to stabilize CaBPs expression in all its subdivisions. This nucleus also shows the most differentiated, time-dependent distribution of CaBPs immunoreactivity (especially CB), distinct in its divisions. The differences in the CaBPs immunoreactivity confirm previous reports concerning dissimilar origin and development, and also reflect the diversity of connectivity of the amygdaloid body - the collection of nuclei, considered as one functional integrity.

The claustrum is a relatively large telencephalic structure, situated close to the border of the neo- and allocortical regions. Its neuronal population consists of glutamatergic, projecting neurons and GABA-ergic interneurons, characterized by occurrence of numerous additional biochemical markers. The postnatal development of these latter neurons has not been extensively studied. Revealing the characteristic patterns of colocalizations between selected markers may shed some light on their function and origin. We investigated the colocalization patterns between three neuropeptides: neuropeptide Y, somatostatin, vasoactive intestinal polypeptide and three calcium-binding proteins: calbindin D28k, calretinin, parvalbumin in the interneurons of the rat claustrum during a four-month postnatal period (P0-P120; P: postnatal day). Our studies revealed the following types of colocalizations: neuropeptide Y with calbindin D28k, calretinin or parvalbumin; somatostatin with calbindin D28k; vasoactive intestinal polypeptide with calretinin. Only vasoactive intestinal polypeptide- and calretinin-containing, double-labeled neurons were present at the day of birth, whereas the other double-labeled neurons appeared at later stages of development. The ratios of colocalizing neurons to single-labeled neurons in each type of colocalization were differentiated and reached the highest value (51%) for vasoactive intestinal polypeptide- and calretinin-double-labeled neurons. In conclusion, the claustral interneurons represent differentiated population in respect to the occurrence of neuropeptides and calcium-binding proteins. The expression of studied substances is changing during the postnatal period.

There is large body of evidence suggesting distinct susceptibility to ischemia in various brain regions. However, the reason for this remains unexplained. Comparative studies of programmed cell death (PCD) pathways indicate their differentiated evolutional origin. The caspase-independent pathway is regarded as an older, whereas the caspase-dependent--as more advanced. In our study we address the question of whether there are any characteristic differences in the activation and course of PCD in phylogenetically and morphologically distinguished brain structures after transient focal ischemia. Using Western blot, we studied changes in expression of caspases: 3, 8, 9, and AIF in the frontoparietal neocortex, archicortex (CA1 and CA2 sectors of the hippocampus) and striatum, during reperfusion after 1 h occlusion of the middle cerebral artery. The caspase and AIF expression were differentiated between the studied structures. The activation of only the caspase-dependent pathway was observed in the neocortex. In the archicortex and striatum both caspase-dependent and caspase-independent pathways were activated, although in the latter the extrinsic apoptotic pathway was not activated. In summary, it is conceivable that structures of different evolutionary origin undergo cell-death processes with the participation of phylogenetically distinguished mechanisms. The previously reported unequal susceptibility to ischemia may co-exist with activation of different cell death pathways.

The role of fibroblast growth factor receptors (FGFR) in normal brain development has been well-documented in transgenic and knock-out mouse models. Changes in FGF and its receptors have also been observed in schizophrenia and related developmental disorders. The current study examines a transgenic th(tk-)/th(tk-) mouse model with FGF receptor signaling disruption targeted to dopamine (DA) neurons, resulting in neurodevelopmental, anatomical, and biochemical alterations similar to those observed in human schizophrenia. We show in th(tk-)/th(tk-) mice that hypoplastic development of DA systems induces serotonergic hyperinnervation of midbrain DA nuclei, demonstrating the co-developmental relationship between DA and 5-HT systems. Behaviorally, th(tk-)/th(tk-) mice displayed impaired sensory gaiting and reduced social interactions correctable by atypical antipsychotics (AAPD) and a specific 5-HT2A antagonist, M100907. The adult onset of neurochemical and behavioral deficits was consistent with the postpubertal time course of psychotic symptoms in schizophrenia and related disorders. The spectrum of abnormalities observed in th(tk-)/th(tk-) mice and the ability of AAPD to correct the behavioral deficits consistent with human psychosis suggests that midbrain 5-HT2A-controlling systems are important loci of therapeutic action. These results may provide further insight into the complex multi-neurotransmitter etiology of neurodevelopmental diseases such autism, bipolar disorder, Asperger's Syndrome and schizophrenia.

A growing body of evidence indicates the common origin of the claustrum, endopiriform nucleus, and the basolateral nuclear complex of amygdala from the lateral and ventral parts of the pallium, as the claustroamygdaloid complex. It seems very probable that at least some of the claustral interneurons derive from subcortical sources. The postnatal development of neuropeptide Y-, somatostatin- and vasoactive intestinal polypeptide-containing interneurons was studied during the 4 postnatal months (P0-P120; P, postnatal day). The study was conducted on 45 Wistar rats of both sexes. Our results indicate that neuropeptide-containing interneurons are not morphologically mature at the moment of birth. The characteristic features of neuronal bodies and the relatively long period of postnatal development may indicate their migration from the subcortical neurogenetic centers. Morphological changes in the neuropil are also reported. Although developmental patterns differ between various neuropeptide-containing neuronal subpopulations, two phases of development can be distinguished in each of them: the early phase (P0-P4) during which undifferentiated neurons and neuropil dominate, and the late phase (P7-P28) during which the characteristic features of an adult-like structure gradually appear. Later these observed developmental changes are terminated. The postnatal development of neuropeptide-containing interneurons is completed after 4 weeks of life. This period, which is important for the structural and functional development of the claustrum, must be taken into account in future studies on this structure.

The piriform cortex (PC), the primary olfactory cortex, is involved in the processes of learning and stress response and possibly plays an important role in epileptogenic activity. The results of several recent studies suggest that those PC neurons that contain neuronal nitric oxide synthase (nNOS) may play a key role during spatial learning and in the modulation of initiation, propagation and generalisation of seizures in various experimental models and may influence neuronal vulnerability after epileptic insults. The aim of this study was to characterise the pattern of distribution and morphology of nNOS-immunoreactive elements in PC of the adult rabbit. The co-localisation of nNOS and calretinin (CR) was also studied. The pattern of nNOS-ir within the rabbit PC is similar to that described previously in other mammals. The morphology of nNOS-ir elements, namely varicose fibres and Cajal-Retzius cells, suggest that NO has an important influence on PC function. Surprisingly, in the rabbit PC nNOS-ir elements show a very low level of co-localisation with CR-ir.

Spontaneous intracerebral haemorrhage carries a high mortality rate and treatment of the disease raises more questions then answers. Mass effect, ischaemia and toxicity of blood components are responsible for brain tissue damage. Initially occurring disturbances of cerebral blood flow have a temporary character and do not play a key role in the pathology of intracerebral haematoma. Oedema formatting in the 24-48 hours after intracerebral bleeding is the result of multidirectional processes. The pathological mechanism that underlines it is the function of activation of systemic complement and cascade of coagulation. In the light of these findings, further clinical and experimental investigations should be focused on these factors.

Background: Microglial cells play an important role in the pathophysiology of intracerebral haemorrhage. We have examined the possible influence of sevoflurane on the reactivity of microglial cells during intracranial haemorrhage., Methods: Forty adult male rats were divided into two groups. All animals were anaesthetized with fentanyl, dehydrobenzperidol and midazolam. In the experimental group animals additionally received sevoflurane 2.2 vol% end-tidal concentration. Intracranial haemorrhage was produced through infusion of blood into the striatum. The microglial cell population (numerical density of immunoreactive cells and their distribution) was assessed on days 1, 3, 7, 14 and 21 after producing a haematoma using antibodies OX42 and OX6., Results: In the control group significant differences in the density of OX42-ir cells between 3rd and 7th (81.86 vs. 129.99) (95% CI: -77.99 to -18.25, P = 0.0035) and between 14th and 21st (105.36 vs. 63.81) (95% CI: 13.21 to 69.89, P = 0.006) survival days were observed. However, significant increase of percentage of amoeboid OX42-ir cells between 3rd and 7th (0.98 vs. 48.71) (95% CI: -52.17 to -43.30, P = 0.0001) and between 7th and 14th (48.71 vs. 58.47) (95% CI: -13.96 to -5.55, P = 0.0002) and then their decrease - between 14th and 21st (58.47 vs. 31.74) (95% CI: 22.52 to 30.93, P = 0.0001) days of observation were noted. In the sevoflurane groups OX42-ir cells were not found. On the 3rd day the density of OX6-ir cells in the sevoflurane group was significantly lower than that in the control group (12.39 vs. 34.57) (95% CI: -49.78 to -2.96, P = 0.02). The percentage of an amoeboid form of OX6-ir cells was significantly lower in the sevoflurane group than that in the control group (27.31 vs. 82.03) (95% CI: -72.52 to -36.92, P = 0.0001) (58.76 vs. 82.37) (95% CI: -38.81 to -8.41, P = 0.003) (42.87 vs. 81.55) (95% CI: -53.23 to -24.10, P = 0.0001) respectively for 3rd, 7th and 14th days of survival., Conclusion: Administration of sevoflurane during anaesthesia in animals with intracerebral haemorrhage evoked a decrease of activation of the microglial cells.

The amygdala is a critical component of the neuroanatomical stress circuit. It plays a role in the generation of responses to emotional stimuli. The central (CeA) and medial (MeA) amygdaloid nuclei are implicated in activation of the hypothalamic-pituitary-adrenocortical (HPA) axis. The immunoreactivity (-ir) of c-Fos, NGF and its receptor, TrkA, following acute and chronic open-field stress were studied in the CeA and MeA nuclei of the amygdala. The material consisted of 21 male adult rats divided into three groups: non-stressed (control) animals, rats exposed to acute (once only lasting 15 min) and chronic (15 min daily over 21 days) aversive stimulation (open-field exposure). The brains were stained with the use of immunohistochemical methods for c-Fos, NGF or TrkA. In the control rats c-Fos-, TrkA- and NGF-ir cells were observed in the nuclei studied, but the quantity varied, being moderate or high (immunoreactive to TrkA and NGF) or low (immunoreactive to c-Fos). In the animals exposed to acute open-field stress the number of c-Fos-ir, NGF-ir and TrkA-ir cells in the nuclei under examination was differentiated but higher than that in the control animals. In the animals exposed to chronic open-field stress the number of c-Fos-ir cells in the nuclei studied was similar and was smaller than those in animals exposed to acute stress. The number of TrkA-ir neurons was also lower in comparison to that in animals exposed to acute stress. However, no significant differences in the number of NGF-ir cells were observed between the groups exposed to acute and chronic stress. Diverse expression of c-Fos protein following both acute and chronic stress stimulation may prove the functional heterogeneity of the amygdaloid nuclei investigated. The decrease observed in both c-Fos- and TrkA-ir in MeA (only TrkA in CeA) of animals exposed to chronic stress may indicate the phenomenon of habituation.

The aim of our study was to analyze the influence of the open field (OF) exposure on: 1. Distribution of c-Fos positive nuclei in: ventral tegmental area, substantia nigra, periaqueductal gray. 2. Appearance of calbindin-D28k, calretinin and parvalbumin in midbrain neurons that are engaged in the stress response. 3. Changes of c-Fos and calcium-binding proteins expression during maturation. The material consisted of Wistar rats of age between 0 and 90 days. The OF exposure was applied throughout 10 min and 90 min before the death of the animals. The brain sections were double stained using the antibodies against c-Fos, CB, CR or PV. Our results showed that in all studied nuclei age-related increase of c-Fos expression (without changing of its distribution properties) was found. PV didn't show any co-localization with c-Fos in neurons of studied regions at any ages, however some PV-immunoreactive (PV-ir) basket-like structures around c-Fos-immunoreactive (c-Fos-ir) neurons were observed. In the youngest group of rats c-Fos-ir cells and cells immunoreactive for CB and CR constituted separate neuronal populations. During maturation increases in the level of their co-localization with c-Fos was observed. We may conclude that in adult rat midbrain structures CB-immunoreactive (CB-ir) and CR-immunoreactive (CR-ir) cells (probably projection neurons) are mainly activated in the stress response following OF exposure. In the contrary PV-ir cells has only an indirect (modulatory) influence upon the c-Fos-ir cells.

The immunoreactivity (ir) for c-Fos, NGF and TrkA, following an acute and chronic open field stress, were studied in the periventricular zone of rat hypothalamus. Adult rats were divided into three groups: control, exposed to acute (single exposure--15 minutes) and chronic (multiple exposures--15 minutes daily for 21 days) open field stress. In the control rats neurons immunoreactive to c-Fos, TrkA and NGF were found. The number of TrkA- and NGF-ir cells was high, whereas this of c-Fos-ir ones was low. In animals exposed to acute open field stress the number of c-Fos-ir cells in the examined nuclei varied, however it was much higher than that in the control animals. The number of TrkA-ir neurons in all the studied nuclei was also higher than that in the control animals, but the increase of the number of NGF-ir neurons was not observed in supraoptic nucleus. In the animals exposed to chronic open field stress the number of c-Fos-ir cells was increased in comparison to that in the control rats. After chronic stress exposure the number of TrkA-ir neurons in supraoptic nucleus remained high in comparison to that in animals exposed to acute stress, whereas it was decreased in other studied nuclei. No significant differences in the number of NGF-ir cells were observed between the groups exposed to the acute and chronic stress. Observed decrease of c-Fos- and TrkA-ir in the studied nuclei in the animals suffering from chronic stress in comparison with the acute one may indicate the occurrence of habituation phenomenon. This phenomenon does not concern NGF-ir.

Claustrum is a telencephalic structure integrating information of various modalities. Proper functioning of this structure depends on the presence of a network of intrinsic connections. This includes GABA-ergic neuronal populations that also contain calcium-binding proteins (CaBPs). The goal of this study was to analyze qualitative and quantitative the 5-HT-containing fibers in the rat claustrum and to assess the relationships between these fibers and the populations of claustral neurons expressing CaBPs. We used the methods of immunocytochemistry and morphometry. The serotonergic fibers in the claustrum are heterogeneous, both with respect to their morphology and spatial distribution. Thin varicose fibers are more numerous and are homogeneously distributed within the claustrum. Remaining fibers were thicker and possessed larger varicosities. They were present mainly in the ventral part of the claustrum. Although the serotonergic fibers are found in the vicinity of claustral cells containing CaBPs, direct contacts between these fibers and cells are rare. Other mechanisms, including volume transmission, may possibly mediate serotonergic influences.

40 adult Wistar rats were divided into two groups depending on the applied anaesthesia. In both groups animals were generally anaesthetized with fentanyl, dehydrobenzperidol administered intraperitoneally and midazolam given intramuscularly. In the second group (SEVO) animals received sevoflurane of 2.2 vol% end-tidal concentration. Intracerebral haematoma was produced through infusion of 100 microl of autologous blood into the striatum. Each group was divided into five subgroups depending on the length of survival period: 1, 3, 7, 14, 21 days. The astrocytic population was studied by means of anti-GFAP staining. Stereological analysis was applied to estimate the numerical density of immunoreactive cells and the distribution of their types. On 7th day of observation the density of GFAP-immunoreactive astrocytes in SEVO was lower (p<0,05) than that in the control group. In the control group, the increase (p<0.05) of per cent of activated astrocytes between the 1st and 3rd survival day was noted, which remained at this level till the end of observation. In SEVO group, the increase (p<0.05) of per cent of activated astrocytes between the 3rd and 7th day and the decrease (p<0.05) between the 14th and 21st survival day were observed. During days of observation the per cent of activated astrocytes was lower (p<0.05) in the SEVO group than that in the control group. Administration of sevoflurane during anaesthesia to animals with intracerebral haemorrhage has evoked not only the delay of the activation of astrocytes but also decrease in its level.

Recently the increasing prevalence of gastrointestinal diseases, including neoplasm, has resulted in the necessity of characterising not only the tumours, but also healthy mucosa. Research into the morphological changes of healthy mucosa under different experimental conditions, including drugs, special diets and the use of probiotic bacteria, is greatly facilitated by the availability of animal models. In spite of the widespread use of mice in gastrointestinal research, there is a lack of information on the qualitative and quantitative histological characteristics of the intestinal mucosa of the mouse. The aim of this study was to assess the morphological characteristics and the postnatal development of the small intestine of wild type mice -- C57BL/6J. The mice were aged either 5 weeks or 12 weeks. The 12-week-old mice had been weaned at the age of 5 weeks. After dissection the small intestine was divided into 5 equal portions and randomly chosen microscopical sections from each were stained with haematoxylin and eosin. The parameters describing the morphology of the small intestine (villus height, depth of the crypt, villus width near the crypt, width of the villus connective tissue near the crypt, thickness of the muscular layer and the height of the enterocytes and their nuclei) were evaluated under a light microscope. In both age groups the height and width of the villi decreased, while the thickness of the muscular layer increased in the distal direction. The height of the enterocytes decreased and the height of the enterocyte nucleus increased towards the colon in both age groups. The depth of the crypts was greater in the younger animals than in the older ones. Our data provides the baseline morphological description of the small intestinal mucosa in wild type mice, strain C57BL/6J, which can be used as a reference for testing the influence of drugs, toxins, nutrients and inborn mutations on the mouse intestine.

In our study we used c-Fos protein (as a marker of cellular activity) to identify whether cells containing parvalbumin (PV) in the piriform cortex (PC) are engaged in the response to stress stimulation and to discover how this expression changes during maturation. The material consisted of Wistar rats of postnatal (P) ages between 0 and 120 days divided into 9 groups: P0, P4, P7, P10, P14, P21, P30, P90, P120. Each group consisted of 5 experimental and 3 control animals. Rats of the experimental groups were exposed to the "open field test" throughout 10 minutes. The control animals were kept in a home cage. Our results showed that c-Fos activity in the open field test was observed in layers II and III of PC after birth. It then increased and stabilised on P30. In the second week of life PV-positive cells were also observed in those layers. These achieved maturity in the 4th week of life. After this time basket-like structures appeared but the level of PV/c-Fos co-localisation was low. Only small differences were observed between the anterior and posterior parts of PC. In the anterior part a higher number of PV-positive neurons, neuropil threads, and basket-like structures and a larger degree of PV/c-Fos co-localisation were observed. Our results suggested that during maturation PV cells are not directly activated in response to stress stimuli but PV neurons via their numerous endings influence the activation of c-Fos-positive cells predominantly in the anterior part of PC.

A total of 14 patients of various ages diagnosed with schizophrenia and, as an age-matched control group, 12 healthy subjects were examined using the MRI method of neuro-imaging. The volume of the following structures was evaluated in the right and left hemispheres: the superior temporal gyrus, the basolateral temporal area (the region including the middle temporal gyrus, inferior temporal gyrus and fusiform gyrus), the parahippocampal gyrus, the hippocampal head, the amygdaloid body and the inferior horn of the lateral ventricle. In schizophrenia a significant increase in the volume of the amygdaloid body on both the left and right sides was observed. In the patients, as in the control group, we noticed significant asymmetry between the left and right sides in the volume of the structures studied. The left amygdaloid body was significantly larger than the right, whereas the left hippocampal head and the temporal horn of the lateral ventricle were smaller than the right. Our findings suggest that in the early stages of schizophrenia, despite the increased volume of the amygdaloid body, the asymmetry between the structures of the temporal lobe is still present. However, the changes observed in the temporal lobe could be related to the functional disturbances observed in this disease.

Our intention in the present study was to ascertain whether NO-producing cells in the basolateral complex (BLC) and paracapsular intercalated nerve cell groups (Ip) of the amygdala are activated in the open field (OF) test. The material consisted of 8 adult rat brains. The OF test was applied throughout 10 min and 90 min before the death of the animals. The brain sections were double stained using the antibodies against c-Fos (marker of neuronal activation) and against nitric oxide synthase (NOS -- marker of NO-producing cells). The neurons containing NOS and those revealing c-Fos activity constituted distinct populations within both the BLC and Ip but NOS-immunoreactive fibres often surrounded the c-Fos-immunoreactive neurons. Our results suggest that (1) neurons of the basolateral complex of the amygdala and paracapsular intercalated islands are involved but probably not crucial for the open field stress response and (2) NOS-immunoreactive cells in the BLC and Ip are not activated after OF exposure.

The developmental changes of 25-kDa synaptosomal-associated protein (SNAP-25) expression in the rat striatum and cerebral cortex were examined using Western- blotting and densitometric scanning of immunoblots. Analysis of the striatum extracts from postnatal day 0 (P0) to postnatal day 120 (P120) demonstrated that SNAP-25 is poorly expressed until P14. From this point the expression level gradually increases to reach a maximum on P60 and then decreases. The pattern of SNAP-25 expression in the rat cerebral cortex is different. Two peaks are observed, the first on P10 and the second on P60, after which the expression level decreases. These results appear to confirm the role of SNAP-25 protein in axon outgrowth and synaptogenesis in the nervous system.

The endopiriform nucleus, further divided into dorsal and ventral parts, and the neighbouring pre-endopiriform (pEn) nucleus form a region of highly heterogeneous structure involved in numerous physiological and pathological processes. Nonpyramidal neurons of this region containing three neuropeptides-somatostatin (SOM), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP)-were examined in this study. Their colocalization with three calcium-binding proteins-parvalbumin (PV), calbindin D28k (CB), calretinin (CR), and with nitric oxide synthase (NOS), was investigated by qualitative and quantitative methods. The results are summarized as follows: (1) all studied substances are distributed in neurons of the entire region, (2) SOM-ir neurons constitute the most numerous neuropeptide-containing population, whereas NOS-ir neurons make up the largest population of all studied, (3) colocalizations are found in the endopiriform region (Enr) (SOM with CB, PV and NOS; VIP with CR; NPY with NOS and NOS with CR), (4) heterogeneity of the endopiriform region appears in the differences of cells' shape distributions of single-labeled (SOM-, CR-PV-ir) and double-labeled (SOM/CB-, SOM/PV-, NPY/NOS- and NOS/CR-ir) neurons, as well as in differentiated percentage values of SOM/NOS, NPY/NOS and VIP/CR double-labeled neurons in three studied parts; additionally, differences in distribution of immunoreactive neuropil elements between parts of the region are observed. Numerous regional differences concerning neuronal morphology and immunocytochemical characteristics justify further division of the endopiriform region into distinguished parts. Some immunocytochemical features of the neurons in studied region may contribute to the role in epileptogenesis.

The piriform cortex has been extensively studied due to its possible role in epileptogenic activity. Neurones containing calcium-binding proteins (CaBPs), as a component of inhibitory circuitry, seem to be critically involved in this pathological process. The aim of the present study was to characterise the pattern of distribution of CaBPs-immunoreactivity in the piriform cortex of the adult rabbit. It comprises labelled cells, fibres (often with varicosities) and terminals. It varies among the layers. Moreover, the distribution of the parvalbumin- and calretinin-immunoreactive fibres and terminals allows even further subdivision of the layer I into two sublayers. Calretinin-ir neurones are located in subpial (Ia) layer, while parvalbumin - as well as calbindin-D28k-ir ones are mainly located in the second and third layer. Cajal-Retzius-like neurones containing calretinin, Chandelier cells containing parvalbumin and basket cells containing calbindin D28k and parvalbumin can be distinguished among labelled subpopulations of CaBPs neurones. In general, the pattern of PV- CR- and CB-immunoreactivity is similar to that previously characterised in other mammals, i.e., rats, guinea pigs, hedgehog, and tenrecs. The pattern is organised in topographic fashion confirming the complexity of regulatory circuits in the rabbit piriform cortex.

The present paper describes parvalbumin, calbindin-D28k and calretinin immunoreactivity in the claustrum and endopiriform nucleus of adult rabbits. Studied neuronal populations are characterized by morphological heterogeneity. Four types were identified in each subpopulation of cells containing calcium binding proteins on the basis of the number of processes and their branching pattern. There were no spatial differences in the distribution of cells containing either parvalbumin or calbindin-D28k in the claustrum and endopiriform nucleus. Well documented presence of the various projective zones in the rabbit claustrum did not reflect the specific distribution of neurons containing calcium binding proteins, except those containing calretinin. Their localization may correspond with the limbic zone. We have found that the rabbit claustrum and endopiriform nucleus have different pattern of parvalbumin and calretinin immunoreactivity. The former was more intense in the claustrum and the distribution of cell types was significantly different from that in the endopiriform nucleus. Calretinin-positive cells were observed in the claustrum, while in the endopiriform nucleus they were scarce. The distribution of neither calbindin-D28k-ir neurons nor fibers allowed differentiation of claustrum and endopiriform nucleus. Significant differences between the claustrum and endopiriform nucleus observed in the rabbit might be related with ontogenetic as well as other (functional) factors.

Depression is one of the most common psychiatric disorders and is associated with considerable morbidity. In recent years structural-imaging technology has provided an opportunity to examine the brain anatomy in patients with the psychiatric illness. 10 patients of various ages and, as the control group, 16 healthy subjects were examined using the MRI method of neuroimaging. The volumes of the following structures were evaluated in the right and left hemispheres: the superior temporal gyrus, the basolateral temporal area (the region including middle temporal gyrus, inferior temporal gyrus and fusiform gyrus), the parahippocampal gyrus, the hippocampal head, the amygdaloid body and the lateral ventricle. The significant difference between the control group and the group with depression concerned the volume of the temporal horn of the lateral ventricle of both hemispheres. In depressed patients the left temporal horn was 49.8% and the right 38.4% larger in comparison with the control group. In the control group there were significant differences between the left and right hemispheres in the volume of all the structures studied, whereas in the group with depression these difference in volume between the hemispheres concerned only the amygdaloid body and the lateral ventricle.

The morphometric analysis of changes occurring in the rat and rabbit ventroposterolateral (VPL) nucleus of the thalamus during the postnatal development was performed using unbiased stereological methods. The materials used in the study included 30 Wistar rats and 32 New Zealand rabbits aged from P0 to P180 (P-postnatal day), which were divided into six and eight age groups, respectively. The following stereological parameters of VPL nucleus on the cresyl violet stained sections were determined: volume of the nucleus, numerical density and total number of neurons. The total number of neurons indicated that the development of VPL nucleus in both species ended within the third week of postnatal life. The volume of VPL nucleus increased gradually (by about 2.2 and 5 times in rats and rabbits, respectively) in comparison with the volume of the cerebral hemisphere during the development from P0 to adulthood. The numerical density of VPL neurons decreased rapidly at the beginning of postnatal life and stabilized by the end of the third week. In both species, the gradual increase in the volume of VPL nucleus and the simultaneous decrease in the neuronal density in the first week of postnatal life were mainly caused by changes in the neuropil volume. The total number of cells did not change remarkably during the first postnatal week. However, it decreased significantly during the second week. This decrease was probably due to the naturally occuring cell death. These results show that the most prominent qualitative and quantitative changes in VPL nucleus and its neurons occur during the first two weeks of postnatal life of rats and rabbits. Also, because the thalamocortical relay neurons completely acquire their physiological features, this the most critical period of time for their morphological maturation.

A comparative quantitative study of the somatosensory thalamocortical connections in the rat and rabbit, labeled with the fluorescent retrograde tracer Fluoro-Gold (FG), was conducted by means of unbiased stereology. FG was injected into the primary somatosensory cortex of the rat and rabbit in different age groups from P0 to P180 (P-postnatal day). The numerical density of retrogradely labeled the ventroposterolateral (VPL) projection neurons was analyzed. A significant decrease in this parameter was observed during the first two weeks of postnatal life in both studied species. Changes of the neuropil volume and selective elimination of early cortical connections stemming from the VPL may possibly cause this process. A withdrawal of axon collaterals from the expanded cortical sites as well as apoptosis (existing both in the VPL and parietal cortex) contribute to a decrease in the numerical density. Our observations allow us to conclude that the thalamocortical somatosensory connections established before the birth undergo significant quantitative changes in both studied species during the first two weeks of postnatal life and this period seems to be crucial for maturation of the thalamocortical loop.

The morphological features of nitric oxide synthase (NOS)-containing neurons in the rat claustrum (Cl) were studied during the period of four months after birth. Forty-five animals divided into nine groups, according to survival period (P0, P4, P7, P10, P14, P21, P28, P60, P120) were used in the study. The immunocytochemical staining to neuronal NOS was performed and the material was studied both qualitatively and quantitatively using unbiased stereological methods. Our observations indicate that the process of maturation of NOS-immunoreactive (ir) neurons in Cl takes place during the early postnatal period. We report the increase of numerical density of immunoreactive neurons, changes in neuronal size, expressed by the decrease of the percentage of small neurons with simultaneous increase of the participation of medium-sized neurons and large neurons. In the whole studied period the prevalence of oval and fusiform neurons is observed. However, the increase of the proportion of multipolar neurons takes place. Round neurons are most characteristic in the youngest groups of animals and later become dominated by the developing subpopulations of ir neurons of other shapes. In the anterior, central and posterior parts of Cl, a similar pattern of maturation of NOS-ir neurons is observed. No prevalence of characteristically shaped neurons is observed in any part of Cl. The adult-like pattern of morphological features in the NOS-ir neuronal population in Cl is reached in the third postnatal week. The maturation of NOS-ir neurons in the claustrum is a dynamic process which is not stabilised at the moment of birth. It may be assumed that characteristic changes of the NOS-ir population of neurons may be influential on the physiological processes observed in Cl. These may in particular have some importance for the processes of synaptogenesis and establishing as well as refining of numerous claustral connections with the other structures of the central nervous system.

Intracerebral haemorrhage is a strong stimulus for both microglial and astroglial activations. There are some important pathophysiological features during haemorrhage that do not occur in ischaemic or traumatic brain injuries, and may influence the dynamics and intensity of glial activation. Studies on the evolution of glial reaction may have practical importance to the introduction of new therapeutic methods for influencing the inflammatory reaction during haemorrhage. Microglial and astroglial responses to experimental intracerebral haematoma were studied in 50 adult rats for 5 minutes after injection of 100 microl autologous arterial blood into the striatum. The survival period varied from 1 to 21 days. Microglial-macrophage lineage cells were immunocytochemically stained with antibodies OX42, OX6 and ED1. The astrocytic population was studied by means of anti-GFAP staining. Changes in cellular morphology and intensity of staining were time-dependent reactions in both microglial and astroglial cells. Strong activation of microglial-macrophage lineage cells revealed with OX6-and OX42-immunoreactivity started during the first postoperative day. The complete pattern of activation for ED1-immunoreactivity was observed from the third postoperative day. At this stage, numerous phagocytic macrophages started to appear in the perihaematoma region. Morphological changes were most intensive during the second postoperative week. The astroglial (anti-GFAP) reaction was observed after the third postoperative day and proceeded less dynamically. The glial reaction gradually stopped but not completely during the period of observation. The early occurrence of glial activation, pattern of morphological changes and characteristic sequence of antigens expression indicate a very intense type of glial reaction. Evolution of glial response to haemorrhage reveals characteristic features. In our opinion, the initial phase of glial activation, comprising 72 hours after the occurrence of haemorrhage, is potentially the most promising period for influencing the extent of glial reaction with therapeutic agents.

An immunocytochemical double-staining method was applied in order to study the co-localisation of nitric oxide synthase (NOS) with three calcium-binding proteins, calbindin D28k (CB), calretinin (CR) and parvalbumin (PV) in the claustrum of the rat during the first 4 months of life (postnatal days: PO-P120). The co-localisation of NOS/PV and NOS/CB is reported. These neurons fall into the category of non-pyramidal cells. Double-labelled NOS/CB neurons are observed in the claustrum starting from P4, whereas double-labelled NOS/PV neurons are observed from P14 onwards. The percentages of double-labelled neurons increase in relation to the age. Double-labelled NOS/CB and NOS/PV neurons, although they do not constitute a numerous population, play an important role in the process of maturation of the claustrum. This is confirmed by the occurrence of these types of neurons at definite stages of maturation and by the increase in their number.

The aim of the present study is to follow topographical and morphological changes in the development of the amygdaloid basolateral complex (BLC) in the rabbit. The material consists of 35 brains of New Zealand rabbits of both sexes, divided into 7 age groups (P2-P90). In cresyl violet preparations BLC is already well visible on P2 and is composed of the lateral (divided into dorsolateral and ventromedial divisions), basolateral and homogenous basomedial nuclei. On about the 7th postnatal day it is possible to divide the basomedial nucleus (BM) into dorsal (Bmd) and ventral (BMv) divisions. The topography and subdivisions set on P7 are maintained in further periods of life. The morphology of neurons (shape, dendrites, staining) changes significantly until P21 in all BLC nuclei. Our results indicate that BLC achieves morphological maturity relatively late, which is probably connected with a long creation of emotional memory and regulation of emotional behaviour.

A properly structured neuronal network, which is a prerequisite of a correctly functioning central nervous system, depends on the proper construction of synaptic terminals. There are some studies concerning development of hippocampal cells, however, the data about maturation of the synaptic terminals network within the hippocampal formation are only fragmentary. The study was performed on Wistar rats of various ages. The following synaptic proteins: synaptophysin, SNAP-25 and GAP-43 (a growth axon cone protein) were used to study the maturation of synaptic terminals. We have found that, at the time of birth, the hippocampal formation is very immature. Synaptogenesis begins on P4 and lasts till P10 or P14. The strongest immunoreactivity of all proteins is observed at the beginning of this period. Thereafter, immunoreactivity decreases and, after P14, it is similar to that of adult animals. On P60 and P90, the level of immunoreactivity increases again. In the pyramidal and granular cell layers, the formation of synaptic terminals and their maturation occur earlier in embryonal life, and are almost completely mature at birth.

The morphometric analysis of changes occurring in the rabbit claustrum in the early postnatal period was performed by means of unbiased stereological methods. Material consisted of 40 animals aged from P2 to P180 (P-postnatal day) divided into eight groups. The volume of the claustrum, total number and numerical density of its neurons change very rapidly at the beginning of the postnatal life and stabilize by about the fourth week. From the 21st postnatal day distribution and morphology of neurons correspond to that in adults. Almost from the beginning of the postnatal life the rabbit claustrum is composed of three different parts: anterior, central and posterior. The anterior part is the largest and it consists mainly of fusiform, pyramidal and multipolar neurons. The central part is generally composed of oval neurons. The posterior part is the smallest; the distribution of its neuronal types is similar to that in the anterior one. Rapid morphological changes of the claustrum and its neurons occurring during the first postnatal month seem to point out that this structure is able to fulfill its physiological role after this critical period.

We compared the volumes of the superior temporal gyrus, basolateral temporal area, parahippocampal gyrus, hippocampal head, amygdaloid body and the inferior horn of the lateral ventricle in 29 patients with dementia of Alzheimer's type (DAT) and in 14 cognitively normal controls. Measurements were performed on coronal MR images perpendicular to the long axis of the hippocampus then the raw data were normalised for intracranial volume. The volumes of all studied structures were significantly smaller in patients with DAT than in age-matched control group. The differences in the volume between DAT and age-matched control group were the largest for the amygdaloid body and the basolateral temporal area. The discriminant analysis including the volume of both the basolateral area and the left amygdaloid body allows for correct classification of 100% DAT patients and 93% controls. We conclude that the estimation of the volume of the amygdaloid body and basolateral temporal area seems to be the most important factor for DAT diagnosis.

The aim of the study was to assess the effect of storage time on the actual thickness of the histological sections. The study was performed on 5 brains of adult Wistar rats. The most rapid changes occur at the beginning of the storage process, after about one month the dynamics of changes decreases, but still the thickness of the sections diminishes. It is suggested that quantitative analyses should be performed in similar period of time and the critical care should be taken not only of the chemicals and procedures used, but also of the control of environmental factors.

The aim of the study was to assess the effect of age of the animal upon the real thickness of the frozen sections. The study was performed on 19 rabbit brains. The thickness of the frozen sections regardless of their staining is age-dependent. The relation is proportional during the period from 7 to 180 postnatal day and characterizes both immunohistochemical as well as cresyl violet-stained sections; moreover, changes of the section thickness proceed parallelly. It is suggested that especially for some stereological parameters all required procedures should be standardized to achieve comparable and unbiasedly interpretable results.

The aim of the present paper is to describe the morphology and topography of the nuclei of the amygdaloid complex in the rabbit. In the current study we also investigated the intensity of the enzymatic reaction for acetylcholinesterase (AChE) in the amygdaloid complex and the morphology of its neurones. Material consisted of 5 brains of adult New Zealand rabbit, stained either with cresyl violet or for AChE activity. Although, as in other mammals, the rabbit amygdala consists of two main nuclear groups (corticomedial and basolateral), it reveals a peculiar morphology pattern, forming a transition structure between those observed in the cat and rat. Especially characteristic is the arrangement of the basolateral complex. Within that the ventromedial division of the lateral nucleus seems to be the largest, while its dorsolateral division--the smallest. The arrangement of the corticomedial complex in the rabbit is similar to both the cat and rat. In the rabbit the highest acetylcholinesterase activity is found in the basolateral nucleus and the nucleus of the lateral olfactory tract. The lowest AChE staining is observed in the cortical and medial nuclei, amygdalohippocampal and anterior amygdaloid areas and intercalated masses.

The aim of this study of the cerebral cortex of 8 non-demented elderly subjects and of 17 subjects in the severe stage of Alzheimer's disease (AD) (Global Deterioration Scale stage 7/Functional Assessment Staging procedure stage 7a-f) was to examine the relationships between amyloid-beta (Abeta) deposits and neurofibrillary degeneration. The study shows that neuronal processes with neurofibrillary changes are detectable in only a minority of fibrillar plaques: from 31% to 49% of fibrillar plaques within frontal, temporal, parietal, limbic, occipital, and insular cortices. The correlations observed between the numerical densities of neurons with neurofibrillary tangles (NFTs) and the densities of Thioflavin-S-positive fibrillar plaques with neurofibrillary changes (r=0.61; P<0.01) indicate that neurofibrillary pathology in neocortical plaques reflects the topography and rate of neurofibrillary changes in neocortical neurons. The accumulation of abnormally phosphorylated tau in only some plaques indicates that fibrillar Abeta enhances paired helical filament accumulation locally only in dystrophic neurites already involved in neurofibrillary degeneration. The lack of correlation between the number of neurons with neurofibrillary changes and the number of all Thioflavin-S-positive fibrillar plaques (with and without neurofibrillary changes) suggests that beta-amyloidosis does not contribute to initiation of neurofibrillary degeneration in neurons.

Background: Sevoflurane is one of the newest volatile anaesthetic agents. The effect of sevoflurane on ICP in conditions of intracranial pathology is essential from the clinical point of view but still not sufficiently clear. The aim of the study was to examine the effects of 1 MAC, 2 MAC, 3 MAC sevoflurane on intracranial pressure (ICP), mean arterial blood pressure (MABP) and heart rate (HR) in rabbits with experimental intracerebral haematoma (ICH)., Material and Methods: The experiments were performed in 13 adult rabbits, 3.5-4.0 kg weight. The rabbits were randomly allocated to two different groups. In group I, (n = 7), sevoflurane was administered in stepwise increasing concentrations of 2.2 vol%, 4.4 vol% and 6.6 vol%, each for a period of 15 minutes. In group II (n = 6), intracerebral haematoma was produced and subsequently sevoflurane was administered in the same manner as in group I. Ventilation parameters, inspiratory and end-tidal sevoflurane concentration, end-tidal CO2 concentration (ETCO2), HR, MABP, ICP and body temperature, measured in the nasopharynx, were monitored throughout the experiment., Results: Mean values of ETCO2 and temperature in the nasopharynx were not significantly different in both groups and remained stable in the whole observation period. In group II in all cases the evidence of intraventricular haematoma was observed. In this group mean values of ICP, MABP and HR after haematoma production were significantly higher than those in group I. Statistically significant increase of ICP was observed in 30th minute in group I, while in 35th minute in group II. In both groups a statistically significant decrease in MABP was observed from 20th minute of observation. A significant decrease in HR in both groups from 25th minute of experiment was also observed., Conclusion: In conclusion it should be stressed that sevoflurane, in doses not exceeding 1 MAC, shows no significant effect on ICP and cardiovascular function in the course of intracranial haematoma.

A morphometric study of amyloid-beta-positive plaques in the neocortex of eight non-demented people from 68 to 82 years of age and 17 subjects with late-stage Alzheimer disease (GDS stage 7/FAST stages 7a-f) from 73 to 93 years of age shows a shift from prevalence of fibrillar plaques to prevalence of nonfibrillar plaques. In the aged, non-demented subjects, about 4/mm^2 plaques are detectable in the neocortex, and the majority are fibrillar plaques. Specifically, 64% found to be classical fibrillar and Thioflavin-S-positive bright primitive plaques. A lower percentage of pale primitive plaques (35%) relatively small proportion of plaques that are poor in thioflavin S-positive fibrils. The numerical density of plaques in the severe stage of AD increases to about 41/mm^2. Severely demented subjects appear to maintain an active process of fibrillar plaque formation. This is reflected in the presence of 3% bright primitive plaques. Severely demented subjects also manifest plaque degradation, reflected in the presence of 22% and 48% percentages of classical fibrillar plaques in non-demented subjects and in the end stage of disease suggest that once activated, the process of fibrillar plaque formation persists at a somewhat stable rate during the whole course of brain amyloidosis.

Dementia of Alzheimer type and vascular dementias account for as much as 90% of all dementias. MRI volumetry of the temporal lobe is often performed in Alzheimer's disease, but there are only a few studies that evaluate temporal lobe atrophy in vascular dementia. We therefore compared volumes of the superior temporal gyrus (STG), basolateral temporal area (BTG--the region including middle temporal gyrus, inferior temporal gyrus and fusiform gyrus), parahippocampal gyrus (PAH), head of the hippocampus (HIP), amygdaloid body (AA) and temporal horn of the lateral ventricle (LV) separately for each hemisphere in 8 patients with ischaemic vascular dementia (IVD) and in 14 cognitively and neurologically normal subjects. In the left hemisphere basolateral temporal area, parahippocampal gyrus, amygdaloid body and hippocampal head had significantly smaller volumes in IVD patients than in age-matched control group. In the right hemisphere the significant atrophy in IVD patients concerned only basolateral temporal area.

The purpose of the study was to evaluate the usefulness of magnetic resonance (MR) in imaging of the cisternal and petroclival segments of the abducent nerve. Heavily T2-weighted submillimetric 3D sequence in axial plane, T1-weighted 3D, 1.5 mm slice thickness sequence in axial plane and TOF sequence were performed on 16 volunteers. Additionally the reformatted T2-weighted images in sagittal and in oblique parasagittal plane parallel to the abducent nerve in the pontine cistern were performed. The heavily T2-weighted sequence provides high contrast between fluid and other structures. High signal intensity of the cerebrospinal fluid is a kind of background for cranial nerves, vessels, meninges as well as for bony and fibrous structures. The authors identified the abducent nerve in at least one plane of the submillimetric, heavily T2-weighted sequences in 84.4% cases (in 84.4% in axial plane, 68.8% in sagittal and 84.4% parasagittal parallel to the VI-th cranial nerve in the pontine cistern). Dorello's canal was identified in 27/32 abducent nerves (84.4%) on the submillimetric, heavily T2-weighted sequence in parasagittal parallel to the abducent nerve in the pontine cistern plane. In 71.9% (23/32) of cases, the abducent nerve was in contact with the arterial vessel in pontine cistern.

The quantitative analysis of the claustrocortical connections in the rabbit, labeled with the fluorescent retrograde tracer Fluoro-Gold (FG), was conducted by means of unbiased stereology. The FG was injected into selected regions of the motor, somatosensory, auditory and visual cortices and then a comparison of the various claustrocortical projections was carried out. This was achieved by comparing (1) the numerical densities of projecting neurones for each claustral projection zone and (2) the distribution of the labeled neurones throughout the rostro-caudal extent of the claustrum. No significant differences between the numerical densities of labeled neurones in the various projection zones are reported. The motor and primary somatosensory projections dominated in the anterior and central parts of the claustrum, whereas the secondary somatosensory, auditory and visual projections--in the posterior part. The difference in the distributions was significant (p < 0.001). Summarizing, the cortical projections in the claustrum, although varying topographically, do not reveal a quantitative differentiation. This may speak in favour of the integrative and modulating function of this structure in relationship to the neocortex.

Diet and fluoride can modify tooth-germ development. In many morphological and biochemical studies malnutrition was shown to impair odontogenesis. However, quantitative observations of the morphological changes implemented by underfeeding and fluoride are still scanty. The aim of the study was to assess stereologically the enamel and dentin deposition in tooth-germ of 14-day-old rat pups derived from dams fed with deficient diet and given water without or with low (10 mg/l) and high (110 mg/l) doses of natrium fluoride, starting from the 13th day of pregnancy. The volume fractions of ameloblasts, enamel, dentin and odontoblasts in histological sections were estimated by the point counting method. The lack of fluoride in drinking water in rats maintained on low-protein diet changed the proportions of the deposited dental mineralised tissues as compared to the control animals: it substantially increased deposition of enamel (by 48%), and significantly decreased dentin production (by 28%). The supplementation of drinking water with fluoride in rats fed with deficient diet partially reversed these effects towards values found in the control rats maintained on standard diet that drank water with trace amount of fluoride. The possible toxic activity of high doses of fluoride can only be conferred to the decreased volume fraction of ameloblasts. Our findings suggests an important role of the fluoride ion in the maintenance of the proper enamel and dentin relation in the developing teeth of rats fed with low-protein, low-fat, high-carbohydrate diet.