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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. more...

The rat is the most frequently used animal in scientific inquiry conducted for the purpose of advancing basic knowledge that may lead to an improvement in the results of treatment. Understanding of the pharmacological properties of inhalation anaesthetics, in combination with monitoring of their concentration in the inspired and end-tidal gas, together provide safe and precise control of the depth of the anaesthesia. However, accurate application of the inhalation method of anaesthesia requires special equipment for the delivery and effective scavenging of inhalation anaesthetics. more...

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. more...

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. more...

The glial cells play an important role in pathophysiology of the intracerebral haemorrhage (ICH). Thus the attempt at evaluating the possible influence of the propofol on the reactivity of astro- and microglial cells in the course of ICH was performed. 50 rats were divided into two groups depending on the applied anaesthesia. All animals were generally anaesthetized with fentanyl, dehydrobenzperidol and midazolam. No additional agents were given to the animals of the control group (group I). In the experimental group (group II), the animals received additionally intraperitoneally propofol in a dose of 50 mg/kg every thirty minutes. ICH was produced through infusion of the blood into the striatum. The astrocytic and microglial cells population was assessed on the 1, 3, 7, 14 and 21 days after producing a haematoma using antibodies anti-GFAP and OX42. The stereological analysis was applied to estimate the numerical density of immunoreactive cells and the distribution of their types. On the 14th and 21st days of observation the density of GFAP-immunoreactivity (ir) cells was significantly higher in group II than that in group I. There were no differences in percentage distribution of GFAP-ir astrocytes between group I and group II. On the 3rd, 14th and 21st days of observation the density of OX42-ir cells was higher in group II in comparison with group I. For the 7th, and 21st days of survival the percentage of the ameboid form of OX42-ir cells was significantly lower in group I than that in group II. The administration of propofol during anaesthesia in the animals with ICH has evoked an increase of the activation of the astro- and microglial cells. more...

Intracerebral haematoma was produced in 25 adult rats by infusion of 100 microl of autologous blood into the striatum. The animals' brains were removed at 1, 3, 7, 14 and 21 days after production of the haematoma. The TUNEL method was used to detect DNA fragmentation and TUNEL-positive cells were qualified. TUNEL-positive cells were already found on the first day of observation and were present for three weeks after haematoma production. These results provide evidence that programmed cell death is associated with intracerebral haemorrhage. more...

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. more...

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. more...

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. more...

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. more...

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. more...

We examined the protein expression of the calcium-binding protein calbindin D28k in two developing rat brain structures, the striatum and the cerebral cortex. The relative protein concentration level was quantified by means of the Western blotting method and densitometric scanning. 32 Wistar rats were used, divided according to survival period (P0-P120-postnatal days). Observations of the calbindin D28k protein expression in the rat striatum and the cerebral cortex revealed an increase in band color intensity between P0 and P10. The intensity of protein staining in older groups of animals stabilised at a similar level and in the P28 and P120 groups we observed a decrement of calbindin expression in the striatum. Calbindin D28k stabilises the intracellular calcium level, preventing calcium-induced apoptotic cell death in neurons. Thus, changes in calbindin D28k expression might be related to its neuroprotective role in differentiation and synaptogenesis during the postnatal development of the brain. more...

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. more...

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. more...

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. more...

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. more...

We examined the expression of brain nitric oxide synthase (bNOS) in two developing rat brain structures, the striatum and the cerebral cortex. For this purpose, we quantified the relative protein concentration level using the Western blotting method and densitometric scanning. 32 Wistar rats, divided according to survival period (PO-P120-postnatal days) were used in this study. Our results demonstrate that bNOS expression rises in these structures during the first week of postnatal life, reaching a maximum in the striatum on the 10th day and in the cerebral cortex on the 7th day of postnatal life. After the period of increase the expression declines and after the 14th day a stabilisation of bone protein concentration is observed, both in the striatum and the cerebral cortex. These changes in bone protein expression might be related to the important role of nitric oxide in the developing rat brain, especially in synaptogenesis, apoptosis and neurotransmission. more...

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. more...

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. more...

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. more...

Nitric oxide (NO) is a short-lived radical, which modulates synaptic plasticity, neuronal oscillations and cerebral blood flow. NOS-containing neurones can be detected anatomically by nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry or by NOS immunohistochemistry. Neuropeptide Y(NPY) is the most abundant peptide in the brain. NPY is connected with several vital functions, such as a feeding behaviour, sexual maturation, regulation of circadian rhythms, body temperature, blood pressure and neuroendocrine secretions. Neuropeptide Y also modulates anxiety-related disorders, limbic epileptic seizures as well as learning and memory processes. The study was performed on 45 Wistar rats of various ages (PO, P4, P7, P10, P14, P21, P30, P60, and P120; P--postnatal day). The free-floating sections were stained with standard immunohistochemistry methods. Thereafter the histological sections were studied using the confocal laser microscope equipped. For 3D reconstruction the image analysis program LaserSharp 2000v. 2.0 (Bio-Rad, UK) was used. We found that in the newborn rat both NOS- and NPY-immunoreactivity was weak. It had been increasing gradually until the 7th day of postnatal life, after that until P14 it was maintained on the similar level, and then the number of immunolabelled cells deceased. The developmental changes concerned cell morphology as well--until the 10th day of life the immunoreactive cells were immature, with round or oval bodies and had only a few fibres. From P14 the cells' morphology became similar to that in adult. more...

The rat is one of the species most commonly used in laboratory practice. Numerous publications concerning various aspects of morphology and physiology are based on the results obtained in this species. It make these results comparable and under some precautions enables to transpose into the relationships observed in humans. Each experimental project must obtain the permission of the Local Ethical Committee, as well as comply with the regulations of the European Communities Council, outlined in the "European Convention for the protection of vertebrate animals used for experimental and other scientific purposes". Adequate pre-operative care can eliminate or reduce the incidence of many complications, which may occur during anaesthesia. General anaesthesia in experimental practice can be achieved using a variety of drugs and ways of administration, among others inhalational or intravenous. The side effects of anaesthetic agents can be reduced in this way. Knowledge of the effect of anaesthetics on the cerebral circulation, metabolism and intracranial pressure in both normal and pathological conditions is crucial for neurobiological purposes. Many anaesthetic agents depress respiration, which can result in hypoxia, hypercapnia and acidosis. To maintain blood carbon dioxide and oxygen concentration in the physiological range, it is necessary to apply tracheal intubation and artificial ventilation. However, even when using sophisticated equipment, the role of basic clinical observation, such as the colour of the blood shed in the operation field, breathing depth and frequency, cannot be overestimated. The importance of monitoring mean arterial blood pressure and intracranial pressure in experiments on the central nervous system is fundamental. Special attention should be paid to controlling the temperature and monitoring the fluid balance. Appropriate postoperative care can have a decisive influence on the final results of the research. more...

Retrograde axonal transport method of the fluorescent tracer FluoroGold (FG) was combined with immunocytochemistry to investigate the occurrence of nitric oxide synthase (NOS), somatostatin (SOM), neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) in both intrinsic and cortically projecting neurons of the rat claustrum. Only NOS was detected in both the scattered projecting neurons and internal neurons of the claustrum. Approximately 20% of NOS-immunoreactive neurons in the claustrum were also retrogradely labeled with FG after tracer injections into the frontal cortex. The other substances were exclusively confined to the population of interneurons, which mainly displayed an oval, round or fusiform shape and a medium size. Apart from the neuronal somata, the proximal parts of the dendritic arborization were clearly visible. The immunoreactive neurons were randomly distributed in the claustrum and their neuronal size and shape did not differ in the various parts of the studied structure. Co-localization of NOS and SOM or NOS and NPY was reported. In conclusion, SOM, VIP and NPY do not appear to play a significant role in the claustro-cortical projection but are most probably involved in modulation and information transfer in the claustrum. The appearance of NOS in both cortically projecting and intrinsic neurons of the claustrum may be indicative of a fundamentally different role in the functioning of the claustro-cortical loop. more...

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. more...

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. more...

The experimental model of the intracerebral hematoma in the rabbit was used for the investigation of the changes of the intracranial pressure and selected hemodynamic parameters. The study was performed on 13 adult rabbits, divided into two groups receiving 1 ml (group I--6 animals) and 2 ml (group II--7 animals) of fresh arterial blood, respectively. The monitoring of the intracranial pressure (ICP), the mean arterial blood pressure (MABP), the heart rate (HR), the end-tidal CO2 concentration (ETCO2) and the body temperature was measured every minute in the hematoma production phase and every 5 minutes for the consecutive three hours. The volume of the hematoma was calculated according to Cavalieri formula, with the use of the system for the automatic picture analysis. The mean volume of the intraparenchymal part of the hematoma in group I was higher than in the group II. However, in all the representatives of the second group the evidence of the intraventricular hemorrhage was present. The dynamics of the ICP, MABP and HR changes differed significantly in both groups during the period of the observation. On the basis of the physiological and morphological observations we conclude that the changes of ICP remain the most sensitive and valuable parameter during the early course of the intracerebral hemorrhage. Coexistence of the rapid ICP, MABP and HR changes must be always regarded as the possible sign of the intraventricular hemorrhage. more...

Using axonal retrograde tracing, combined with morphometric analysis, we compared the distribution and number of claustral neurons projecting to the motor and somatosensory cortical areas in the Wistar rat. Comparable volumes of the retrograde tracer Fluoro-Gold, were injected into the motor or somatosensory cortices. Injections into these areas resulted in labeling of neurons along the entire length of the claustrum. Neurons retrogradely labeled after injection into the motor cortex prevailed in the anterior part of the claustrum, whereas those projecting to the somatosensory cortex predominated in the central part. The mean number of claustral neurons retrogradely labeled after tracer injections into the motor cortex significantly outnumbered that from the somatosensory cortical area (p < 0.01). Similarly, the mean value of the numerical density of the retrogradely labeled neurons was significantly higher for the motor projection zone in the claustrum, than for the somatosensory projection zone (p < 0.001). The contralateral claustral projections, both into the motor and somatosensory cortices, were considerably lower in number than the ipsilateral ones. These findings indicate that: (1) the claustral projections to the various cortical regions seem to be differentiated (2) the distribution of claustral neurons projecting to the motor and somatosensory neocortical areas shows an anteroposterior gradient, (3) the claustrum of the rat appears to be more closely related to the motor than to the somatosensory system, (4) the rat claustrum seems to function more as a satellite than a relay structure in relationship to the cerebral cortex. more...

The connections between two parts of the claustrum in the rat and rabbit were studied using the highly fluorescent lipophilic carbocyanine dye (Dil). After the application of Dil crystal into the endopiriform nucleus, labeled fibers in the insular claustrum were observed in its part directly neighboring the insular cortex and capsula externa. Additionally, numerous projections into the piriform, insular and entorhinal cortices were present. The presence of connections between the endopiriform nucleus and insular claustrum suggests its role concerned with the processes taking part in the allocortical regions as well as in the limbic system. more...

The quantitative analysis of the claustrocortical connections labeled with the fluorescent retrograde tracer Fluoro-Gold (FG) was conducted on 90 rabbits subdivided into the following age groups (P2, P7, P14, P21, P30, P60, P90, P120, P180). The equal volumes of retrograde fluorescent tracer FluoroGold (FG) were injected into the selected regions of the motor or somatosensory cortices. The volume of the dorsal part of the claustrum, total number of projecting neurons, numerical density and percentage distribution of projecting neurons were estimated by means of the unbiased stereological methods. The claustrocortical connections both with the motor and somatosensory areas in a rabbit are established in the postnatal life. The parts of the claustrum occupied by the motor and somatosensory projection zones as well as the morphology of the cortically projecting neurons do not reveal characteristic changes during the studied period. The significant decrease of the total number and numerical density of cortically projecting neurons as well as the increase of the claustral volume may reflect the process of adjustment of the claustrum to its modulatory function upon corresponding cortical areas. The intensity of the claustral connections with the motor and somatosensory cortices reveals significant difference during the studied period, being higher for the motor projection. It may be assumed that the claustrocortical connections established before birth undergo significant quantitative changes during postnatal development. more...

Unbiased stereological methods were used for estimating the numerical density and the total number of claustral neurones projecting to the cingulate cortex in rabbit and rat. In rat the numerical density of neurones projecting to the retrosplenial granular cortex (RSG) differed significantly from those projecting to the retrosplenial agranular (RSA) and cingulate (Cg) cortices while in rabbit the numerical densities of retrogradely labelled neurones in the claustrum following injections into various areas of the cerebral cortex did not differ significantly. The total number of retrogradely labelled neurones in the claustral limbic zones did not differ significantly in both species. The quantitative analysis of claustral zones projecting to a different cingulate cortex area, both in rabbit and rat, reveals that each of these zones is rather homogeneous. more...

The morphology of the claustrum was studied by stereological methods in representatives of five mammalian orders (Insectivora, Rodentia, Lagomorpha, Carnivora and Primates). In each species under study, a dorsal and a ventral part of the nucleus can be distinguished. Based on differences in shape and separation from surrounding structures, five morphological types of the claustrum occur. The claustrum of Insectivora and some rodents represents the least complicated morphological type. The nucleus is very poorly separated from the surrounding structures. The human claustrum is morphologically the most complicated, although the two above-mentioned principal divisions are apparent. The ventrally situated paraamygdalar part of the human claustrum may correspond to the endopiriform nucleus or ventral part of the claustrum of other mammals, because of its morphological characteristics and connections with the limbic system. In guinea pigs, traditionally classified as members of the Rodentia, a characteristic morphological type of the claustrum is present. This observation may support arguments questioning the current position of this species in mammalian classification. Based on stereological studies, the increase of the claustral volume that occurs with increase of the hemispheric volume is significantly smaller than the increase of the isocortical volume and larger than the increase of the allocortical volume. The increase of the volume of the dorsal and ventral parts of the claustrum does not differ significantly in the species under study. Neurons of the claustrum represent differentiated morphology. The numerical density of neurons in the dorsal part of the claustrum is significantly higher than in the ventral one. Differences in the morphology and cellular structure of the two parts of the claustrum may suggest differences in function of the two parts of the nucleus, most probably concerned with transfer of information among various cortical regions. Changes in the claustrum, a cortico-related structure, that occur with increased brain volume, may suggest that its development is less dynamic than that of the isocortex. more...

The afferent and efferent connections of the piriform cortex and the endopiriform nucleus in the rat were studied by the method of axonal transport of two fluorescent tracers: Fluoro-Gold and Fluoro-Ruby. The results indicate that both structures possess not only the connections with the olfactory system, but also the reciprocal connections with the limbic system (entorhinal cortex, amygdaloid body), thalamus (mediodorsal and midline nuclei), extrapyramidal system (ventral part of the nucleus accumbens). The topographic differences in the organization of the association connections between the anterior and posterior parts of the piriform cortex are reported. Additionally, the reciprocal and relatively numerous connections between the endopiriform nucleus and the piriform cortex may result in their modulatory function, which in some pathological circumstances may have a critical significance in epileptogenesis. more...

The claustrocortical connections in the rabbit were assessed for the first time by the method of axonal retrograde transport of two fluorescent tracers (Fast Blue and Diamidino Yellow). The material consisted of 23 adult New Zealand rabbits. Projection zones of spindle-like form, connected with the precentral, postcentral, temporal and occipital cortices have been delineated. They are organized topographically both in the anteroposterior and ventrodorsal direction. The precentral (motor) projection zone is localized in the anterodorsal part of the claustrum. It may be divided into two separate parts that project to the medial and lateral part of the precentral cortex. The large postcentral (somatosensory) zone occupies mainly the central part, whereas the temporal (auditory) and occipital (visual) zones are situated in the posteroventral part of the claustrum. The overlap of various claustral projection zones is differentiated, the largest being that of the somatosensory zones. In comparison to the results of study of claustral projection zones performed on other species, presumably on the rat and cat, its seems plausible to conclude that the extension of claustral projection zones and degree of their overlap in the rabbit represent an intermediate character. more...

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. more...

Synaptophysin is an integral membrane protein associated with small, electron-lucent synaptic vesicles. Immunohistochemistry for this protein is a sensitive method to study subtle changes in synaptic density and distribution in various brain regions. In the present study, the synaptogenesis was examined in the rat basolateral amygdala in comparison with the hippocampal hilus, from the day of birth to adulthood. A total of 41 brains at various ages starting from P0 to P90 (P--postnatal day) were examined. After perfusional fixation the brains were frozen and cut in the coronal plane and stained either with cresyl violet or standard immunohistochemical methods using the anti-synaptophysin antibody. Synaptophysin positive granules appeared just after birth in both structures, but their number was very low (about 0.28 x 10(6) and 0.13 x 10(6) per mm3 in the amygdala and hippocampus, respectively). In the basolateral amygdala the number of synapses increased rapidly reaching the maximum at P14 (1.6 x 10(6) per mm3) followed by about 45% decrease in number up to P30 and later being stabile. In the hippocampus two increases of the synaptogenesis were observed. The first at P7 (about 1.7 x 10(6) of synapses per 1 mm3) which was followed by dramatic decrease up to 0.7 x 10(6) per mm3 at P14. The second increase appeared later (about P90) and reached 1.7 x 10(6) per mm3. After that time the density of synapses was stabile. It may be supposed that the first characteristic wave of synaptogenesis observed in the hippocampus and amygdaloid body is due to the overproduction of synapses observed at that time in other cortical regions. The late wave of synaptogenesis found in the hippocampus is related to the great plasticity of the interneuronal connections in this period of development. more...

The thalamic reticular nucleus (Rt) is a thin lamina of cells, through which thalamocortical and corticothalamic fibers pass. It is interposed between the thalamic nuclei and the internal capsule and it is composed of GABA-ergic cells with synapses that receive impulses from both kinds of fibers. Rt takes part in the negative feed-back system of controlling the information transfer from the thalamus to the cerebral cortex and it is focused in the sleep-waking cycle. The pattern of parvalbumin reactivity during maturation of Rt becomes the main aim of our study. The study was performed on 36 rats on various postnatal days (P0, P1, P2, P4, P5, P7, P10, P14, P17, P21, P30 and P90). The animals were anesthetized, transcardially perfused, cut on cryostat into 30-microns-thick frontal sections, stained immunocytochemically using standard ABC method and a mouse monoclonal antibody against parvalbumin. A small amount of round and oval, parvalbumin immunopositive cells was detected at stage PO, predominantly in the intermediate part of Rt, whereas the cells in ventral and lateral part at the same time were only slightly immunopositive. At P10 the cells in the intermediate part became more fusiform or oval because of the appearance of dendrites. At P14 we were able to observe separate, punctuated structures interpreted as the axonal endings. There were plenty of them at the time of full maturation of the intermediate portion of reticular nucleus (stage P21). At this time, the dorsal and ventral parts had their first synapses, too, but their maturation ended a week later. At P30 multipolar neurons, with round and fusiform somata were distributed relatively homogeneously throughout Rt. We compared the stage with the parvalbumin reactivity of the adult rat and found no difference in the morphological pattern of PV neurons. more...

The connections of the amygdaloid body with the areas of the limbic cortex in the rat were studied by means of the method on the axonal retrograde transport of the fluorescent tracer FluoroGold. The tracer was injected into the anterior and posterior limbic cortices (cingulate gyrus, granular and agranular retrosplenial area, respectively. The localization of the corresponding amygdalar projection zones was investigated and the semiquantitative analysis of the connections was conducted. The projection zones in the rat amygdaloid body are organized topographically. Administration of the fluorescent tracer to the anterior and posterior part of the limbic cortex in the rat (cingulate gyrus and both retrosplenial areas, respectively) reveals labeling of cells only for injections to the former one. The labeled cells were present only in the major components of the basolateral amygdaloid complex. The main source of this projection was anterior part of basolateral nucleus (BLA). Some labeled neurons were found in the lateral nucleus and few in the ventral part of basolateral nucleus. No labeled cells were found within the basomedial nucleus. more...

We have examined the development of rat striatum for evidence of cells dying in the process of physiological cell death. In present study we have indicated apoptotic cells in sections stained with cresyl violet (cell death characterized by pyknosis) or with DNA end labeling assay (TUNEL method). Our results demonstrated that cell loss during maturation of the rat striatum had the characteristics of apoptosis rather than necrosis. The greatest number of TUNEL-positive and pyknotic cells in the striatum were found during the first postnatal days; after 7th day of postnatal life a rapid decrease of its number was observed after the second postnatal week no TUNEL-positive cells were observed in the striatum. Our analysis suggests that apoptotic cell death occurring during the development of striatal neuronal population takes place during the first week of postnatal life. more...

The corticoclaustral connections in the rat were investigated by means of the method of the retrograde axonal transport of the fluorescent tracer (Fluoro-Gold; FG). The material consisted of 20 adult Wistar rats. The fluorescent tracer was injected into the anterior, middle or posterior parts of the claustrum. The retrogradely labeled neurons were detected in the layer VI of the neocortex. Injections of the tracer into the anterior part of the claustrum resulted in labeling of the neurons in the motor cortex. After administration of the tracer into the middle part of the insular claustrum, labeled neurons were present both in the motor and somatosensory cortices, while the injections of the tracer into its posterior part resulted in labeling of neurons in the visual cortex. Administration of the fluorescent tracer into the insular claustrum of the rat resulted in labeling of the cortical neurons of the corresponding areas of both hemispheres, however, the contralateral projections seem to be less numerous than the ipsilateral. Our results confirm the existence of reciprocal connections of the claustrum with the neocortex and suggest its role in integration and modification of information reaching the neocortex. more...

The authors present two cases of haematoma accompanying coagulopathies, treated successfully in the Department of Neurosurgery, Medical Academy in Gdańsk. In the first case, haematoma occurred after trauma in a patient with haemophilia B, and in the other case in a patient treated with Syncumar for mitral valve implantation. The authors apply the strategy of treatment of patients with coagulopathies which should involve achieving of stable haemostasis, indispensable for positive result of the treatment. more...

A case of extra-intracranial, parietal chondromyxoid fibroma in a 13-year old girl operated successfully and followed by 7-years observation is presented. Apart from infrequent occurrence of the tumour diagnostic difficulties due to atypical cranial location are emphasized. more...