Prolonged cultivation of separated rat lung mast cells (LMC) in vitro is necessary to better investigate a possible role of LMC in different stages of tissue remodeling induced by hypoxia. Rat lung mast cells (LMC) were separated using a protocol including an improved proteolytic extraction and two subsequent density gradient separations on Ficoll-Paque PLUS and a new generation of Percoll, i.e. Percoll PLUS. Instead of usual isotonic stock Percoll solution, an alternative "asymptotically isotonic" stock solution was more successful in our density separation of LMC on Percoll PLUS. Separated cells were cultivated for six days in media including stem cell factor, interleukins IL-3 and IL-6, and one of two alternative mixtures of antibiotics. These cultivations were performed without any contamination and with only rare changes in cell size and morphology. Model co-cultivation of two allogenic fractions of LMC often caused considerable rapid changes in cell morphology and size. In contrast to these observations no or rare morphological changes were found after cultivation under hypoxic conditions. In conclusions, we modified separation on Percoll PLUS to be widely used, altered LMC separation with respect to purposes of long-lasting cultivation and observed some model morphological changes of LMC.

• MeSH
• centrifugace - gradient hustoty MeSH
• financování organizované MeSH
• isotonické roztoky chemie   MeSH
• krysa rodu rattus MeSH
• mastocyty cytologie   MeSH
• oxid křemičitý chemie   MeSH
• plíce cytologie   metabolismus   MeSH
• povidon chemie   MeSH
• separace buněk metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

haracterized by fibrotization and muscularization of the walls of peripheral pulmonary arteries. This vessel remodeling is accompanied by an increase in the amount of lung mast cells (LMC) and the presence of small collagen cleavage products in the vessel walls. We hypothesize that hypoxia activates LMC, which release matrix metalloproteinases (MMPs) cleaving collagen and starting increased turnover of connective tissue proteins. This study was designed to determine whether in vitro hypoxia stimulates production of MMPs in rat LMC and increases their collagenolytic activity. The LMC were separated on the Percoll gradient and then were divided into two groups and cultivated for 24 h in 21 % O2+ 5 % CO2 or in 10 % O2 + 5 % CO2. Presence of the rat interstitial tissue collagenase (MMP-13) in LMC was visualized by immunohistological staining and confirmed by Western blot analysis. Total MMPs activity and tryptase activity were measured in both cultivation media and cellular extracts. Exposure to hypoxia in vitro increased the amount of cells positively labeled by anti-MMP-13 antibody as well as activities of all measured enzymes. The results therefore support the concept that LMC are an important source of increased collagenolytic activity in chronic hypoxia.

• Klíčová slova
• Hypoxia in vitro, Rat lung mast cells, Tissue metalloproteinases, Tryptase, Remodeling of pulmonary vessels,
• MeSH
• financování organizované MeSH
• hypoxie buňky MeSH
• imunohistochemie MeSH
• kolagen metabolismus   MeSH
• krysa rodu rattus MeSH
• kultivační média metabolismus   MeSH
• kultivované buňky MeSH
• mastocyty enzymologie   MeSH
• matrixová metaloproteinasa 13 metabolismus   MeSH
• matrixová metaloproteinasa 2 metabolismus   MeSH
• matrixová metaloproteinasa 9 metabolismus   MeSH
• matrixové metaloproteinasy metabolismus   MeSH
• plíce cytologie   enzymologie   MeSH
• potkani Wistar MeSH
• separace buněk MeSH
• tryptasy metabolismus   MeSH
• upregulace MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Increased blood pressure (BP) in genetic hypertension is usually caused by high activity of sympathetic nervous system (SNS) which is enhanced by central angiotensin II but lowered by central nitric oxide (NO). We have therefore evaluated NO synthase (NOS) activity as well as neuronal NOS (nNOS), inducible NOS (iNOS) and endothelial NOS (eNOS) protein expression in brainstem and midbrain of adult spontaneously hypertensive rats (SHR) characterized by enhanced sympathetic vasoconstriction. We also studied possible participation of brain NO in antihypertensive effects of chronic captopril treatment of adult SHR. NOS activity was increased in midbrain of SHR compared to Wistar-Kyoto (WKY) rats. This could be ascribed to enhanced iNOS expression, whereas nNOS expression was unchanged and eNOS expression was reduced in this brain region. In contrast, no significant changes of NOS activity were found in brainstem of SHR in which nNOS and iNOS expression was unchanged, but eNOS expression was increased. Chronic captopril administration lowered BP of adult SHR mainly by attenuation of sympathetic tone, whereas the reduction of angiotensin II-dependent vasoconstriction and the decrease of residual BP (amelioration of structural remodeling of resistance vessels) were less important. This treatment did not affect significantly either NOS activity or expression of any NOS isoform in the two brain regions. Our data do not support the hypothesis that altered brain NO formation contributes to sympathetic hyperactivity and high BP of adult SHR with established hypertension.

• MeSH
• financování organizované MeSH
• kaptopril farmakologie   MeSH
• krevní tlak genetika   účinky záření   MeSH
• krysa rodu rattus MeSH
• mozek enzymologie   účinky léků   MeSH
• NG-nitroargininmethylester farmakologie   MeSH
• pentoliniumtartrát farmakologie   MeSH
• potkani inbrední SHR MeSH
• potkani inbrední WKY MeSH
• renin-angiotensin systém účinky léků   MeSH
• sympatický nervový systém MeSH
• synthasa oxidu dusnatého, typ I metabolismus   MeSH
• synthasa oxidu dusnatého, typ II MeSH
• synthasa oxidu dusnatého, typ III metabolismus   MeSH
• synthasa oxidu dusnatého antagonisté a inhibitory   metabolismus   MeSH
• tělesná hmotnost účinky léků   MeSH
• velikost orgánu účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

For patients with HD neural transplantation is the only treatment with evidently positive effects, although limited for several years. However, it is still necessary to solve a number of uncleared matters concern. relations between graft and host tissue.Transplantation of foetal neural tissue into neurotoxic (ibotenate) lesion represents suitable experimental model for study of mentioned relations from the standpoint of morphological and behavioural evaluation.

Pro pacienty s HD je neurální transplantace zatím jediným léčebným zákrokem s výrazně pozitivním, i když časově omezeným efektem. Je však třeba ještě dořešit řadu nejasností, týkajících se vztahů mezi transplantátem a tkání mozku příjemce. Transplantacefetální nervové tkáně do neurotické (ibotenátové) léze představuje vhodný experimentální model pro studium zmíněných vztahů z hlediska morfologického i behaviorálního hodnocení.

• MeSH
• corpus striatum MeSH
• Huntingtonova nemoc terapie   MeSH
• modely nemocí na zvířatech MeSH
• transplantace fetální tkáně MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• neurologie
• transplantologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

The neurotoxic lesion of the rat brain, induced by stereotaxic infusion of the ibotenic (IA) or kainic (KA) acids, is a commonly used model of Huntington's disease (HD) in animal studies. Neurodegenerative process in HD develops for 10-20 years. However, the majority of studies related to the animal models of HD deal with only the several weeks surviving animals. For that reason, a detailed description of the development of histopathological changes in the striatum in the rat brain is presented in this study. Intrastriatal instillation of both, the IA or the KA causes the partial necrosis of the striatum, accompanied by a rarefaction of the neuropil. Owing to a rather low number of subsequently in situ proliferating glial cells, predominantly astrocytes, the whole process results in a shrinkage of the striatum compensated by an enlargement of the lateral brain ventricles. Although, the fully developed IA lesion is envisaged at 1 week and KA lesion at 3-4 weeks after the injection of neurotoxic acids, the degenerative process within the striatum develops in a rather long time -- at least 6 months. The only morphological observation that doesn't correlate to the findings from the HD patients, is the needle-track area, which is repaired by a conspicuous glial or glial-fibrotic scar. There is no substantial difference in the histopathological characteristics of both the neurotoxic acids used in our studies. However, if the IA must be applied into 3-4 sites in each hemisphere in comparison with only one injection of the KA, the use of KA is, from the morphological point of view, more suitable in relation to the number of artificial needle-track areas.

• MeSH
• corpus striatum účinky léků   patologie   MeSH
• Huntingtonova nemoc chemicky indukované   patologie   MeSH
• krysa rodu rattus MeSH
• kyselina ibotenová MeSH
• kyselina kainová MeSH
• modely nemocí na zvířatech * MeSH
• neurotoxiny MeSH
• potkani Long-Evans MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH

• MeSH
• Huntingtonova nemoc patologie   MeSH
• krysa rodu rattus MeSH
• modely u zvířat MeSH
• nervová tkáň MeSH
• transplantace metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

• Publikační typ
• abstrakty MeSH

• Publikační typ
• abstrakty MeSH

Aging has been associated with a decay of hippocampal function that may begin well before senescence. Conditioned blocking is a complex learning phenomenon that requires an intact hippocampus in young-adult rats and is absent in middle-aged rats. The aim of the present study was to test the possibility of re-establishing conditioned blocking in 17-month-old Wistar rats by neurotransplantation. Solid embryonic hippocampal or nigral tissue was bilaterally transplanted in the proximity of the dorsal hippocampus (lateral ventricle and alveus). Conditioned blocking of an aversion to a cider vinegar (3%) solution presented in compound with a previously conditioned saccharin solution (0.1%) appeared 14 days after transplantation and persisted 3 months later only in the hippocampal grafted group, showing the possibility of restoring age-related cognitive deficits.

• MeSH
• chuť MeSH
• hipokampus fyziologie   transplantace   MeSH
• krysa rodu rattus MeSH
• kyselina octová MeSH
• paměť fyziologie   MeSH
• podmiňování (psychologie) * fyziologie   MeSH
• potkani Wistar MeSH
• stárnutí * fyziologie   MeSH
• substantia nigra transplantace   MeSH
• transplantace fetální tkáně * MeSH
• transplantace mozkové tkáně * MeSH
• učení vyhýbat se fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• Huntingtonova nemoc chemicky indukované   patologie   MeSH
• krysa rodu rattus MeSH
• kyselina ibotenová toxicita   MeSH
• modely nemocí na zvířatech MeSH
• neostriatum transplantace   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH