Biomedical subjects

J Moryś

Publications and source records attributed to J Moryś.

At least 19 recordsLinked to original sources

PubMed2006

Quantitative analysis of influence of sevoflurane on the reactivity of microglial cells in the course of the experimental model of intracerebral haemorrhage.

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.

Anesthetics, Inhalation↗

PubMed2006

Influence of the "open field" exposure on calbindin D28K, calretinin, and parvalbumin containing cells in the rat midbrain - developmental study.

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.

Animals↗

PubMed2006

The immunoreactivity of c-Fos, NGF and its receptor TrkA after open-field exposure in the central and medial nuclei of the rat amygdala.

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.

Amygdala↗

PubMed2006

Immunoreactivity of c-Fos, NGF and its receptor TrkA in the periventricular zone of the rat hypothalamus after open field exposure.

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.

Animals↗

PubMed2006

The pathophysiology of intracerebral haemorrhage.

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.

Blood Coagulation↗

PubMed2006

The influence of acute and chronic open-field exposure on the hippocampal formation: an immunohistochemical study.

The hippocampus plays a role in new learning, memory and emotion and is a component of the neuroanatomical stress circuit. The structure is involved in terminating hypothalamic-pituitary-adrenocortical (HPA) axis responses to stress and attenuates stress responses by shutting off this axis. The immunoreactivity (-ir) of c-Fos, NGF and its receptor TrkA following acute and chronic open-field stress were studied in CA1-CA3 and the DG of the hippocampus. The material consisted of 21 male adult rats divided into three groups: nonstressed (control) animals and rats exposed to acute (15 min once) and chronic (15 min daily for 21 days) aversive stimulation (open-field exposure). The brains were stained with use of immunohistochemical methods for c-Fos, NGF or TrkA. In the animals exposed to acute open-field stress the number of c-Fos-, TrkA and NGF-ir cells was higher in all the structures studied than in the control animals. However they were differentiated only in c-Fos immunoreactivity. In the rats exposed to chronic open-field stress the number of c-Fos-ir cells in the structures of the hippocampal formation studied was smaller than in rats exposed to acute stress and was comparable to that in the control group. No differences were observed between the groups exposed to acute and chronic stress in the number of TrkA-ir cells in the structures under investigation. The number of NGF-ir neurons in CA1 and CA2 was lower after exposure to chronic than after exposure to acute stress but was still higher than that in the control group. Our findings indicate that neurons of CA1-CA3 and the DG are engaged in the stress response after acute as well as chronic open-field exposure. This is probably related to the important role of the hippocampus in processing new spatial information as well as in the habituation processes, although these appear to have different mechanisms.

Acute Disease↗

PubMed2006

The technique of inhalation anaethesia in experimental investigation in the rat.

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.

Anesthesia, Inhalation↗

PubMed2005

Open field stress and neurons containing calcium-binding proteins in the piriform cortex of the rat.

In the present study we wanted to check whether the expression of the c-Fos protein (the marker of cellular activity) appears in cells containing calcium-binding proteins (CaBPs) in animals exposed to the open field test. Eight adult Wistar rats were examined. In the first step the open field test was applied throughout 10 minutes. After perfusional fixation brains were frozen and cut on the cryostat in the coronal plane and stained with the standard immunohistochemical method. Sections were double stained for c-Fos and CaBPs: parvalbumin (PV), calbindin (CB), calretinin (CR). c-Fos positive cells were localized predominantly in layers II and III of the piriform cortex (PC). The double labeling study showed that neurons containing CaBPs are rarely c-Fos-immunoreactive. Often PV-positive and CB-positive fibers surround c-Fos-positive neurons in layers II and III in a form of a basket. It seems that cells containing CaBPs are not directly involved in the response to aversive stimuli but cells containing those calcium-binding proteins might influence directly c-Fos positive neurons of PC.

Animals↗

PubMed2005

The influence of sevoflurane on the reactivity of astrocytes in the course of the experimental intracerebral haemorrhage in rat.

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.

Anesthetics, Inhalation↗

PubMed2005

Apoptosis in the course of experimetal intracerebral haemorrhage in the rat.

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.

Animals↗

PubMed2004

The relationships between neurons containing dopamine and nitric oxide synthase in the ventral tegmental area.

Ventral tegmental area (VTA) is a heterogeneous group of dopaminergic cells which contains interfascicular (IF), parabrachial (PBP) and rostral linear (RLi) nuclei. Neurons of this area are involved in the regulation of motor and motivational aspects of behavior and reveal high neuronal plasticity. Among many various neurotransmitters and neuromodulators, nitric oxide (NO) is localized in this region. In the present study, we investigated morphology and distribution of nitric oxide synthase (NOS)-positive neurons in VTA and their colocalization with dopaminergic neurons. The study was performed on six adult Wistar rats. After perfusional fixation, the brains were cut, immunostained for tyrosine hydroxylase (TH) and NOS and studied by confocal laser microscopy. In each of the three studied nuclei of VTA we investigated three different neuronal populations. Numerous TH-immunoreactive (TH-ir) and NOS-immunoreactive (NOS-ir) neurons are present in the studied region. Among them, a considerable number showed coexistence of both neurotransmitters. The populations of TH-ir and NOS-ir neurons interact with each other as manifested by the presence of NOS-ir endings on TH-ir neurons and vice versa. Taking the above into account, it may be suspected that NO is involved in the modulation of dopaminergic transmission.

Animals↗

PubMed2003

Postnatal development of the basolateral complex of rabbit amygdala: a stereological and histochemical study.

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.

Acetylcholinesterase↗

PubMed2003

The developmental pattern of c-fos expression in the rat thalamus following open-field stress stimulation.

The thalamic nuclei with their defined set of input-output connections are the primary channel for information flow to the cerebral cortex. Several data suggest that neurons of that area are involved in the response to various aversive stimulations. However the pattern of activation seems to depend on the stress model as well as the stage of maturation. In the present study we would like to check which nuclei of the thalamus show expression of c-fos in the response to the "open field test", and how this response pattern changes during the maturation process. 30 rats of age ranged from P0 to P120 (P-postnatal day) were studied. The experimental group was exposed to the "open field test" for 10 minutes. After perfusion and fixation, brains were cut and stained for c-fos with immunohistochemical method. Our results showed that during development the pattern of c-fos activity in the thalamic nuclei after stress stimulation undergoes significant changes. Distinct c-fos expression was observed in the paraventricular nucleus, intergeniculate leaflet and ventral lateral geniculate nucleus. These findings suggest that these nuclei may play a direct role in the stress reaction involved in the response to the "open field test".

Animals↗

PubMed2002

The influence of storage time upon the real thickness of the histological brain sections.

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.

Animals↗

PubMed2002

Relation between the age of specimen and the shrinkage of brain frozen sections.

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.

Aging↗

PubMed2001

Differentiation in the immunocytochemical features of intrinsic and cortically projecting neurons in the rat claustrum -- combined immunocytochemical and axonal transport study.

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.

Animals↗

PubMed2001

Microglial and astroglial cells in the rat paraclaustral reservoir during postnatal development: an immunohistochemical study.

A immunohistochemical study of postnatal development of the paraclaustral reservoir of migrating cells in the rat brain was performed using anti-GFAP (for astroglia), ED1 and OX-42 (for microglia) antibodies. From birth to the 4th day of postnatal life most GFAP-positive cells in the paraclaustral reservoir are similar to transitional astroglia. From the end of the first postnatal week they have the morphology of mature astrocytes, although during the next week, their density was a slightly higher than in neighboring structures. On the 21st day, the morphology and density of astroglial cells in the ventral part of the external capsule did not differ from the surrounding regions. ED1/OX-42- positive microglial cells present in the paraclaustral reservoir during the first postnatal week represented ameboid microglia; their density was clearly higher than in the neighboring structures. During the second week they began to transform into ramified microglia and from the 21st day on, only OX-42 positive resting microglial cells were observed in the ventral part of the external capsule. We suggest that the paraclaustral reservoir is a place of accumulation of astroglia and microglia during brain development and may possibly serve as source of glial cells for neighboring structures. Alternatively, these glial populations may perform local developmental functions.

Animals↗

PubMed2001

General anaesthesia in rats undergoing experiments on the central nervous system.

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.

Anesthesia, General↗