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Biomedical subjects

D Kluchová

Publications and source records attributed to D Kluchová.

At least 19 recordsLinked to original sources
PubMed2002

Neuronal nitric oxide synthase in the rabbit spinal cord visualised by histochemical NADPH-diaphorase and immunohistochemical NOS methods.

The NADPH-diaphorase (NADPH-d) staining method is widely used in the investigation of both the central and peripheral nervous systems. Neuronal nitric oxide synthase (nNOS) has previously been shown to be responsible for the NADPH-d activity in neurons. However, NADPH-d activity does not always fully represent the enzyme nNOS. We investigated the distribution of NADPH-d activity and nNOS protein in the rabbit spinal cord for all groups of neurons and Rexed's laminae. In most laminae the distribution of NADPH-d activity was identical to nNOS immunoreactivity. Both were present in the dorsal horn and in pericentral areas of the spinal cord, but some differences existed. The superficial part of the dorsal horn (laminae I-III) stained more intensely for NADPH-d than for nNOS. However, the most prominent difference was seen in the lateral part of the dorsal horn--the lateral collateral pathway (LCP). The LCP stained strongly for NADPH-d activity, while nNOS staining was absent. Although there is an excellent correlation between NADPH-d staining and nNOS immunohistochemical staining in the spinal cord in general, the presence of staining differences necessitates the use of immunohistochemistry for some specialized applications.

Animals↗
PubMed2001

Comparative analysis of NADPH-diaphorase positive neurons in the rat, rabbit and pheasant thoracic spinal cord. A histochemical study.

The distribution of NADPH-diaphorase (NADPH-d) activity was investigated and compared in the rat, rabbit and pheasant thoracic spinal cord. The investigation of all spinal cord regions (laminae) in three experimental species revealed marked differences in the distribution of NADPH-d activity. Cross sectional analysis of the spinal cord of the rat, rabbit and pheasant confirmed differences in the shape of the gray matter in all examined species. More detailed investigation of Rexed's laminas showed similar distribution of NADPH-d activity in the spinal cord of the rat and rabbit, which were different when compared with the spinal cord of the pheasant. Ventral horn of the rat and rabbit showed no labelling whereas in pheasant this area possessed a number of scattered, intensively stained neurons. In the location of autonomic preganglionic neurons, differences were found as well. In the rat there was seen a number of densely packed, clearly dark blue coloured neurons. Similarly, these neurons were present in the rabbit spinal cord but they were less numerous. No staining was found in this region of pheasant. Pericentral area (lamina X) and intermediate zone (laminaVII) revealed the presence of NADPH-d positive neurons in all examined species although they differed in number and shape of their bodies. The dorsal horn showed the presence of NADPH-d staining in all three animals but its distribution was different in medio-lateral direction. It can be suggested that observed differencies in the presence and distribution of NADPH-d activity across the examined species may reflect different fylogenetic development.

Animals↗
PubMed2000

Partial colocalization of NADPH-diaphorase and acetylcholinesterase positivity in spinal cord neurons.

The freely diffusible radical, nitric oxide (NO), has been assumed to act as a retrograde signaling molecule that modulates transmitter release. Acetylcholine (ACh) is known to function as a typical neurotransmitter. In the present work we have examined the presence of both transmitters (NO and ACh) and their possible relations in the rabbit spinal cord. In our experiments we have used histochemical methods for the visualization of acetylcholinesterase (AChE) and NADPH diaphorase (NADPH-d) which label neurons that express nitric oxide synthase (NOS). Both histochemical methods were performed separately or together on the same sections of the thoracic spinal cord. NADPH-d positive dark blue stained neurons were seen mostly in superficial and deep layers of the dorsal horn, preganglionic autonomic neurons and pericentral area. The presence of AChE positive amber yellow neurons was confirmed mostly in motoneurons located in the ventral horns and in neurons of the pericentral and intermediate zone. Besides the above mentioned neurons, also double-labeled neurons were found which contained both the yellow and dark blue histochemical product. Their presence was confirmed in the intermediate zone and in the pericentral area. Thus, the co-existence of NADPH-d and AChE occurred in the location of interneurons. Our observations suggest that NO may play a role in the control of cholinergic neuronal activity and that NO can be involved in the modulation of synaptic transmission.

Acetylcholine↗
PubMed2000

Expression of peptidergic and nitrergic structures in dorsal root ganglia of the rabbit.

In this study we have demonstrated the presence of neuropeptide substance P (SP)and nonpeptide neurotransmiter NO (nitric oxide) in the dorsal root ganglia (DRG) of rabbits. NADPH-diaphorase histochemical staining was used for detection of NO and an immunohistochemical method for detection of substance P. A number of DRG cells were stained by SP- and NADPH-d reactions. The presence of SP and NADPH-diaphorase positive cells varied depending upon the spinal level of the DRG. Positively stained neurons were only small and intermediate in size. Cells of large diameter profiles showed no staining. Substance P immunoreactive cells were of brown and dark brown colour, the intensity of NADPH-d staining varied from light to very dark blue. In some DRG cells, there was very significant neuronal co-localization of immunoreactivity for SP and reactivity for NADPH-d. In summary, DRG cells appear to express diaphorase and substance P activity, and some of them show the presence of both neurotransmitters. Recent studies on the participation of NO in the regulation of SP release in the spinal cord suggest, that also in the DRG neurons there may be a close interaction between NO and SP.

Animals↗
PubMed1999

Difference of pericentral NADPH-d positive neurons in the rabbit spinal cord segments.

The purpose of the present investigation was to characterize and determine the number of NADPH-diaphorase positive neurons around the central canal in the rabbit spinal cord. These neurons are known to function as interneurons and are present in all spinal cord segments. They differ in shape of their bodies and in length and branching of their processes. The main differentiation was observed in their number, depending on the place of their localization. The highest number of these NADPH-diaphorase positive neurons was in sacral part (6 in average), the lowest one was noticeable in thoracic spinal cord (1-2 in average). It can be concluded that pericentral neurons of the rabbit spinal cord are capable of synthesizing nitric oxide and that they differ in number, depending on the place of their localization in each spinal cord segment.

Animals↗
PubMed1999

Coexistence of nitrergic and acetylcholinesterase (ACHE)-positive nerve structures of the phaesant spleen.

The coexistence of neuronal NADPH-diaphorase and ACHE activities were investigated in the phaesant spleen by successive double histochemical staining of the same sections. Two types of nerve structures were found in pheasant the spleen: nerve cells and nerve fibres. NADPH-d and ACHE-positive nerve fibres in colocalization enter the spleen in its hilum in the vicinity of splenic artery branches and are gradually distributed in periarterial topography in the white pulp. Only NADPH-d positive nerve cells were seen around the splenic vessels. In the red pulp and splenic capsule, only ACHE-positive nerve fibres were present.

Acetylcholinesterase↗
PubMed1999

The influence of transient spinal ischemia on substance P positive nerve structures.

The aim of this study was to investigate, if transient spinal ischemia and a period of 4-day reperfusion will change the distribution pattern of substance P in the spinal cord of rabbits. Strongly enhanced staining of substance P positive nerve structures appeared in the superficial dorsal horn (laminae I, II), the Lissauer's tract, the pericentral region (lamina X), and in the areas of autonomic nuclei (sympathetic-intermediolateral--IML nucleus and parasympathetic-sacral parasympathetic nucleus--SPN) in the control group. Transient spinal ischemia was produced by occlusion of the abdominal aorta just below the left renal artery. Neuropathology of the lesion 4 days after transient ischemia was characterized by selective necrosis of gray matter in the central part of dorsal horn and medial portions of anterior gray matter. Areas with the most dense accumulation of substance P positive structures stayed almost intact. Therefore, no significant change in the distribution pattern of substance P was found in the spinal cord of animals with ischemia-reperfusion-induced injury.

Animals↗
PubMed1999

Selective sparing of NADPH-d positive spinal cord neurons affected by ischemia.

Histochemical characterization of NADPH diaphorase positive neuronal pools in the rabbit lumbosacral segments was performed during and after transient spinal cord ischemia. Strongly enhanced staining of NADPH diaphorase positive structures appeared in the superficial dorsal horn, the pericentral region and in the neurons of the sacral parasympathetic nucleus at the end of 40 min of abdominal aorta ligation or after 1 day reperfusion. Four days after ischemia, NADPH-d positive neurons and vessels were detected in the central gray matter despite well developed necrosis in this location. Regional nitric oxide synthesis and its vasodilatatory effect during the period of aortic occlusion may account for the observed selective resistance of these spinal cord neurons to transient ischemia.

Animals↗
PubMed1998

[Co-localization of substance P and NADPH-d in the thoracic spinal cord of rabbits].

In this study we have demonstrated the localization of substance P (SP) and nitric oxide (NO), in the thoracic spinal cord of rabbit. SP was concentrated in the dorsal horn of the spinal cord, mostly in the superficial layers (LI, LII), around the central canal (LX), and in the region of intermedia-lateral nucleus (IML nc.). NADPH-d (nicotinamide adenine dinucleotide phosphate diaphorase) is an enzyme, which proves the presence of NO. The highest concentration of NADPH-d positive structures were found in the same regions as the SP-positivity. SP is suggested to play a role in the nociceptive transmission. Localization of NADPH-d revealed also the fact that NO may be involved in nociceptive transmission in the spinal cord. Close association of sympathetic preganglionic neurons and fibers with SP and NADPH-d positive structures suggest a role of these neurotransmitters in the modulation of sympathetic activity. (Fig. 11, Ref. 18.)

Animals↗
PubMed1998

[The effect of local ionizing irradiation on the presence of acetylcholinesterase-positive nerve fibers in the spleen of rats].

Changes in the distribution of ACHE-positive nerve fibres in the spleen of the rat after local irradiation have been studied by light microscopy using direct tiocholin method of El-Badawi and Schenk (1967). ACHE-positive nerve fibers enter the spleen in the vicinity of splenic artery branches and they are gradually distributed in form of perivascular plexuses in the white pulp to the periarterial lymphatic sheath (PALS). One dose of local irradiation of the head of the rat causes changes in the presence and distribution of the ACHE-positive nerve fibres in the spleen of the rat. After an intense decrease or the absence of these fibres during the period of the first days after irradiation, there is an evident restoration of ACHE-positive nerve fibres in the white pulp of the organ. The distribution and topography of these nerve structures is the same as in non-irradiated rats on the 35th day after irradiation. During the following days, there is another evident decrease in ACHE-positive nerve fibres in all described localizations of the spleen. Our findings suggest that the indirect effect of irradiation can affect some changes in innervation of the organ which was not exposed to its direct influence. (Fig. 3, Ref. 13.)

Acetylcholinesterase↗
PubMed1998

[NADPH-positive neural structures in the spleen in pheasants].

This study analyzes results of the presence of NADPH-diaphorase in neural structures of the pheasant spleen. The histochemical method of NADPH-d has been used as a marker of nitric oxide synthase (NOS). The NADPH-d positivity shows the specific sites of nitric oxide (NO), which is considered to be one of the new important neuromodulator. We have demonstrated the presence of NADPH-d positive nerve structures in the spleen of the pheasant. Nerve fibers and nerve cells have shown positively the reaction to NADPH-diaphorase. NADPH-d positive nerve fibres enter the spleen in the vicinity of blood vessels, gradually distributed in perivascular localization in the white pulp, reaching the central artery. NADPH-d positive nerve cells have been localized at the hilus of the spleen around splenic vessels. They were polygonal in shape and they occurred as solitary or numerous cells, forming ganglia. We have not found any NADPH-d positive nerve structures in both the red pulp and capsula. The presence of NADPH-d activity in nerve cells and nerve fibres suggests the utilization of NO in the spleen of the pheasant. (Fig. 4, Ref. 16.)

Animals↗
PubMed1998

[NADPH-diaphorase activity in autonomic preganglionic neurons in the rabbit].

BACKGROUND: In order to provide a morphological basis for the understanding of the role of nitric oxide (NO) in autonomic preganglionic neurons, the present study was designed to clarify the localization and distribution of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity, a marker of NO synthase, in the rabbit spinal cord. METHODS: 11 Chinchilla rabbits of both sexes were used in this study. Animals were kept under conditions of controlled light and heat regimens. NADPH-d activity was examined by histochemical methods. RESULTS: The presence of NADPH-d activity in the spinal cord was detected in the dorsal horn, around the central canal (lamina X) and in autonomic preganglionic neurons. Focused on the latter, there was a prominent NADPH-d activity between T1 and L5 segments in the intermediate zone and less intensive NADPH-d staining between S1 and S3 segments. Differences between the two parts of the autonomic system were seen in the arrangement (periodical in sympathetic, and continuous in parasympathetic), in the length of neuronal processes length (shorter in sacral preganglionic neurons) and in their localization (both are seen in the intermediolateral nucleus, but neurons of the sympathetic autonomic system can be found also medially towards the central canal, whilst those of the parasympathetic system were not). CONCLUSIONS: The present results suggest that NO can be used as a transmitter in preganglionic neurons and can be involved in autonomic and sensory processes. (Fig. 10, Ref. 37.)

Animals↗
PubMed1998

[Quantification of pericentral NADPH-diaphorase neurons in the spinal cord of rabbits].

BACKGROUND: The freely diffusible radical nitric oxide is generated by nitric oxide synthase, and is bioregulatory molecule that functions as a major neurotransmitter. Constitutive nitric oxide synthase exhibits NADPH-diaphorase activity that can be demonstrated histochemically. OBJECTIVE: The purpose of the present investigation was to characterize and determine number of NADPH-diaphorase positive neurons around the central canal in all segments of the rabbit spinal cord. METHODS: Rabbits Chinchilla were used in this experiment. After intracardiac perfusion the spinal cords were removed, cut into slices and histochemical analysis of NADPH-diaphorase activity was performed. Sections were evaluated by using light microscope. RESULTS: NADPH-diaphorase positive pericentral neurons were present in cervical, thoracic, lumbar; sacral and coccygeal segments. They differed in the shape of their bodies and in length and branching of their processes. The main differentiation was observed in their number depending on the place of localisation. The highest number of these NADPH-diaphorase positive neurons was in sacral part (6 in average), the lowest one was noticeable in thoracic spinal cord (1-2 in average). CONCLUSION: Thus, our study suggests that pericentral neurons of the rabbit spinal cord which are capable of synthesizing nitric oxide, differs in number amount depending on the place of their localization in each spinal cord segments. (Tab. 2, Fig. 9, Ref. 21.)

Animals↗
PubMed1997

[Localization of NADPH-d positive structures in the thymus of rabbits and rats].

In order to provide a morphological basis for understanding the role of nitric oxide (NO) in the thymus, present study was designed to clarify the localization and distribution of nicotinamide adenine dinucleotide phosphate-diaphorase (NADP-d) activity, a marker of NO synthase in rabbit and rat thymuses. The NADPH-d histochemistry revealed a different distribution of diaphorase positive reactivity in both animal species, visible as dark blue structures. The rabbit thymus displayed lightly stained cortex, whereas medulla was seen as a rounded complex of intensively stained cells, without sharp demarcation between them. However, unlike the rat thymus, the NADPH-d positive cells appeared to be arranged in an irregular line of dense staining at the corticomedullary junction. In the rat thymus, the NADPH-d positive nerve fibres were not evident, whereas neuronal-like plexuses were seen in the perivascular topography of the rabbit thymus. These results suggest that NO may participate in the regulation of thymic function. (Fig. 6, Ref. 30.)

Animals↗
PubMed1997

How do we teach anatomy?

Two approaches to anatomy teaching have been studied and compared. We have concentrated on the teaching time spent for all parts of anatomy and we have compared external recommendations with the teaching time actually given to the subject at our Faculty of Medicine. In addition to that, various opinions on informational and educational aims in medical anatomy teaching are presented. The comparison mentioned above can be used to select the most useful way in which anatomy may be taught, in the light of current curricular emphases and pressures, and in the context of sound educational principles.

Anatomy↗
PubMed1991

The influence of mechanical injury on the metabolic activity of transplanted cerebral cortex.

The authors studied the metabolic activity of rat embryonic cerebral cortex grafts (ED 15-16) implanted into rat brains immediately (TR0) and 14 days (TR14) after cavity formation. Over a period of two months, the ATP, lactate and glucose concentration in TR0 transplants remained at the same level as observed in the intact cortex, whereas in TR14 transplants the ATP and glucose concentration fell significantly and the lactate concentration rose. The DNA concentration rose in both types of transplants, but the increase was more pronounced in TR0 grafts. Choline acetyltransferase activity (a neuron marker) fell significantly in both cases, but the decrease was greater in TR14 transplants. The results indicate that grafts implanted into the brain immediately after cavities had been formed have better metabolic activity and are capable of longer survival than grafts implanted 14 days after cavitation.

Acetylcholine↗
PubMed1989

Effect of partial ischemia on phospholipids and postischemic lipid peroxidation in rabbit spinal cord.

Rabbit spinal cord, subjected to severe partial ischemia induced by abdominal aorta ligation tightly below the renal arteries, was analyzed for phospholipid composition and levels of lipid peroxidation products after 10, 20, and 40 min of the insult. Under conditions when spinal cord blood flow was decreased below 5% of control, concentrations of inositol and ethanolamine phospholipids were decreased by 30% and 10%, respectively. Phosphatidic acid concentration was also altered during ischemia. No accumulation of thiobarbituric acid reactive substances (TBA-RS), conjugated dienes and fluorescent lipid soluble material was found throughout the ischemic period. Pattern of TBA-RS, conjugated diene, and fluorophore formation during postischemic in vitro incubation without and with a peroxidation couple (Fe2+, ascorbic acid) showed increased susceptibility to postischemic lipid peroxidation in tissues after 20 and 40 min of ischemia.

Animals↗