RESUMO
The use of interspecific crosses in breeding is an important strategy in improving the genetic base of the modern cultivated potato, Solanum tuberosum L. Until now, it has normally been interspecific Solanum hybrids that have been morphologically and cytologically characterized. However, little is known about the genomic changes that may occur in the hybrid nucleus owing to the combination of genomes of different origin. We have observed novel AFLP bands in Solanum tuberosum x Solanum kurtzianum diploid hybrids; 40 novel fragments were detected out of 138 AFLP fragments analyzed. No cytological abnormalities were observed in the hybrids; however, we found DNA methylation changes that could be the cause of the observed genomic instabilities. Of 277 MSAP fragments analyzed, 14% showed methylation patterns that differed between the parental species and the hybrids. We also observed frequent methylation changes in the BC1 progeny. Variation patterns among F1 and BC1 plants suggest that some methylation changes occurred at random. The changes observed may have implications for potato breeding as an additional source of variability.
Assuntos
Genoma , Solanum tuberosum/genética , Animais , Southern Blotting , Quimera , Cruzamentos Genéticos , DNA/genética , Metilação de DNA , Genes de Plantas , Variação Genética , Polimorfismo GenéticoRESUMO
The mechanisms by which sensory nerves elicit dilation of serosal arterioles in the jejunum of rats sensitized to the nematode Nippostrongylus brasiliensis were studied using intravital microscopy. Capsaicin (0.002-2 micrograms), applied as a bolus topically to the serosa, produced a substantially larger dilation in the sensitized rats than in unsensitized rats. Abolition of the primary afferent nerves by neonatal treatment with capsaicin and blockade of capsaicin with the antagonist ruthenium red reduced markedly the dilator actions of capsaicin. Mast cell products are important in the actions of capsaicin, because pretreatment with dexamethasone (4 mg/kg), to eliminate mast cells by a macrophage-dependent mechanism, abrogated the actions of capsaicin. In addition, superfusion of the H1 receptor antagonist diphenhydramine (2 microM) blocked the actions of capsaicin. Neither cyclooxygenase products nor platelet-activating factor was involved in capsaicin-induced dilation. The actions of capsaicin and histamine were mediated via a nitric oxide (NO)-dependent mechanism, because superfusion of an inhibitor of NO synthase (NG-nitro L-arginine methyl ester, 10 microM) blocked their effects. This inhibition of capsaicin-elicited dilation by NG-nitro L-arginine methyl ester was prevented by L-arginine (100 microM), the substrate for NO synthase. Thus the arteriolar dilation evoked by capsaicin activation of primary afferent nerves in N. brasiliensis-sensitized rats involves predominantly the release from mast cells of histamine, which then dilates the vessels by a NO-dependent mechanism.
Assuntos
Capsaicina/farmacologia , Jejuno/irrigação sanguínea , Neurônios Aferentes/efeitos dos fármacos , Nippostrongylus/imunologia , Animais , Arteríolas/fisiologia , Histamina/farmacologia , Masculino , Mastócitos/fisiologia , Neurônios Aferentes/fisiologia , Óxido Nítrico/fisiologia , Ratos , Ratos Sprague-Dawley , Vasodilatação/efeitos dos fármacosRESUMO
Superfusion of capsaicin onto the serosal surface of jejunum of Nippostrongylus brasiliensis-sensitized rats induces a short-lasting (1-3 min), dose-dependent (2 to 20 micrograms) decrease in blood pressure which ranges from -5.3 +/- 1.4% to -22.6 +/- 2.2%. The hypotension evoked by capsaicin was more marked in sensitized rats than in unsensitized animals, which responded only to the highest dose (20 mg) of capsaicin tested. The hypotensive effects of capsaicin were not affected by intravenous injections of mepyramine (10 mg/kg), a histamine receptor antagonist, or by the cyclooxygenase inhibitor indomethacin (10 mg/kg). However, an intravenous injection of a platelet-activating factor (PAF) antagonist, BN 52021 (20 mg/kg), or an intraperitoneal injection of guanethidine (8 mg/kg) 18 h prior to experimentation, to functionally impair the sympathetic nerves, abolished the capsaicin-induced drop in blood pressure. Treatment of neonatal rats with capsaicin reduced by 75% the hypotensive effects of capsaicin, whereas the capsaicin antagonist, ruthenium red, reduced non-significantly the hypotensive action of capsaicin. It is concluded that the activation of jejunal sensory nerves in N. brasiliensis-sensitized rats by capsaicin induced a reflex hypotension that is dependent upon PAF release from mast cells and functional sympathetic nerves. In addition, the afferent function of the sensory nerves are not totally blocked by ruthenium red as capsaicin elicits the reflex hypotension in the presence of this blocker of sensory nerve efferent function.
Assuntos
Capsaicina/farmacologia , Diterpenos , Hemodinâmica/efeitos dos fármacos , Jejuno/inervação , Neurônios Aferentes/efeitos dos fármacos , Nippostrongylus , Reflexo/efeitos dos fármacos , Infecções por Strongylida/fisiopatologia , Anafilaxia/fisiopatologia , Animais , Animais Recém-Nascidos , Pressão Sanguínea/efeitos dos fármacos , Capsaicina/antagonistas & inibidores , Ginkgolídeos , Guanetidina/farmacologia , Indometacina/farmacologia , Jejuno/efeitos dos fármacos , Jejuno/parasitologia , Lactonas/farmacologia , Masculino , Fator de Ativação de Plaquetas/antagonistas & inibidores , Pirilamina/farmacologia , Ratos , Ratos Sprague-Dawley , Rutênio Vermelho/farmacologia , Infecções por Strongylida/parasitologiaRESUMO
1. The effects of two platelet-activating factor (PAF) antagonists, WEB 2086 and BN 52021, in reducing the changes in extravasation (Evans blue technique) and blood flow (radiolabelled microsphere method) to various organs and tissues following anaphylactic shock in the Nippostrongylus brasiliensis-sensitized rat were investigated. 2. Both antagonists attenuated anaphylaxis-induced increases in plasma protein leak in the trachea, stomach and small intestine, although they did not block extravasation in the colon and kidneys. 3. Anaphylaxis-induced decreases in blood flow to the adrenals were effectively antagonized by WEB 2086, although this antagonist did not reverse blood flow decreases to any other tissues. BN 52021, on the other hand, did not alter anaphylaxis-induced decreases in blood flow to the adrenals, but effectively prevented dramatic decreases in blood flow to the large and small bowel and spleen. 4. Anaphylactic shock produced marked reduction in blood pressure that was partly reversed by WEB 2086, whereas BN 52021 effectively blocked the decreases in cardiac output. 5. Thus, PAF is responsible for some of the haemodynamic and extravasation of protein changes associated with systemic anaphylaxis in the rat, although the differential inhibition observed with the two antagonists suggests that PAF alters vascular responsiveness through different mechanisms in selected tissues.
Assuntos
Anafilaxia/fisiopatologia , Diterpenos , Nippostrongylus/imunologia , Fator de Ativação de Plaquetas/fisiologia , Anafilaxia/imunologia , Animais , Azepinas/farmacologia , Proteínas Sanguíneas/metabolismo , Azul Evans , Ginkgolídeos , Hemodinâmica/efeitos dos fármacos , Lactonas/farmacologia , Masculino , Microesferas , Fator de Ativação de Plaquetas/antagonistas & inibidores , Ratos , Ratos Sprague-Dawley , Triazóis/farmacologiaRESUMO
1. Changes in tissue and organ blood flow associated with sensitization of rats to the nematode parasite, Nippostrongylus brasiliensis, were studied 30 to 35 days after infection, a time when very few worms remain in the animal. 2. Neither active nor passive sensitization modified heart rate, mean arterial blood pressure, cardiac output or total peripheral resistance. Passive sensitization and administration of non-immune sera did not modify blood flow to any of the tissues studied. 3. Active sensitization increased hepatic arterial blood flow, but decreased blood flow to the stomach, duodenum, jejunum and the submandibular glands. These effects cannot be attributed to residual nematode infections as treatment with the anthelmintic, thiabendazole, did not alter blood flow relative to untreated, actively sensitized rats. 4. The effects of active sensitization on blood flow were probably due to an action of platelet-activating factor (PAF) since treatment of actively sensitized animals with the selective antagonists, WEB-2086 and BN 52021, reversed the decrease in flow seen to the intestinal regions. The PAF antagonists increased blood flow to the kidneys and the trachea of sensitized animals. 5. These results suggest that the PAF released from undetermined sources in nematode-sensitized rats, produces altered blood flow, primarily to the stomach and proximal small bowel.