<b>Suppl. Fig. 1.</b> Detection of (A) glyceraldehyde 3-phosphate dehydrogenase (Gapd... more <b>Suppl. Fig. 1.</b> Detection of (A) glyceraldehyde 3-phosphate dehydrogenase (Gapdh), (B) clathrin (Cltc), (C) Chrm1, (D) Chrm2, (E) Chrm3, (F) Chrm4, (G) Chrm5, (H) Chat by RT-PCR of pulmonary arterial segments from the South American rattlesnake, <i>Crotalus</i> <i>durissus</i>. The products were sequenced and confirmed detection of the right gene product. (A, B) RT positive samples from 1: Proximal, 2: Distal, 3: Branches, 4: Truncus, 5: Proximal, 6: Branches of the pulmonary artery. (C, D, F, G) RT positive samples from 1: Proximal, 2: Distal, 3: Branches, 4: Truncus, 5: Proximal, 6: Branches of the pulmonary artery, while RT negative (genomic contamination) is checked for the same samples in -1 to -6. (E, H) RT positive samples from 1: Proximal, 2: Distal, 3: Branches of the pulmonary artery, while RT negative (genomic contamination) is checked for the same samples in -1 to -3. Primers can be seen in Table 1.
The South American rattlesnake, Crotalus durissus, has been successfully used as an experimental ... more The South American rattlesnake, Crotalus durissus, has been successfully used as an experimental model to study control of the cardiovascular system in squamate reptiles. Recent technical advances, including equipment miniaturization, have lessened the impact of instrumentation on in vivo recordings, and an increased range of anesthetic drugs has improved recording conditions for in situ preparations. Nevertheless, any animal-based experimental approach has to manage limitations regarding the avoidance of pain and stress the stability of the preparation and duration of experiments and the potentially overriding effects of anesthesia. To address such aspects, we tested a new experimental preparation, the decerebrate rattlesnake, in a study of the autonomic control of cardiovascular responses following the removal of general anesthesia. The preparation exhibited complex cardiovascular adjustments to deal with acute increases in venous return (caused by tail lifting), to compensate for blood flow reduction in the cephalic region (caused by head lifting), for body temperature control (triggered by an external heating source), and in response to stimulation of chemoreceptors (triggered by intravenous injection of NaCN). The decerebrate preparation retained extensive functional integrity of autonomic centers, and it was suitable for monitoring diverse cardiac and vascular variables. Furthermore, reanesthetizing the preparation markedly blunted cardiovascular performance. Isoflurane limited the maintenance of recovered cardiovascular variables in the prepared animal and reduced or abolished the observed cardiovascular reflexes. This preparation enables the recording of multiple concomitant cardiovascular variables for the study of mechanistic questions regarding the central integration of autonomic reflex responses in the absence of anesthesia.
Understanding the basis of vascular tonus regulation is fundamental to comprehending cardiovascul... more Understanding the basis of vascular tonus regulation is fundamental to comprehending cardiovascular physiology. In the present study, we used the recently developed decerebrate rattlesnake preparation to investigate the role of nitric oxide (NO) in the control of vascular tonus in a squamate reptile. This preparation allowed multiple concomitant cardiovascular parameters to be monitored, while avoiding the deleterious effect of anesthetic drugs on autonomic modulation. We observed that both systemic and pulmonary circuits were clearly responsive to NO signaling. NO increased vascular conductance in the systemic and pulmonary systems. Vasodilation by NO of the systemic circulation was compensated by cardiovascular alterations involving venous return, cardiac output, and cardiac shunt adjustments. The cardiac shunt seemed to be actively used for hemodynamic adjustments via modulation of the pulmonary artery constriction. N(ω)-nitro-L-arginine methyl ester injection demonstrated that NO contributes to modulating resting vasodilation in the systemic circuit. In contrast, NO-mediated vasodilation did not have an important role in the pulmonary circulation in inactive decerebrated snakes at 25°C. These responses vary importantly from those described for anesthetized snakes.
American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 2020
Vascular tone in the reptilian pulmonary vasculature is primarily under cholinergic, muscarinic c... more Vascular tone in the reptilian pulmonary vasculature is primarily under cholinergic, muscarinic control exerted via the vagus nerve. This control has been ascribed to a sphincter located at the arterial outflow, but we speculated whether the vascular control in the pulmonary artery is more widespread, such that responses to acetylcholine and electrical stimulation, as well as the expression of muscarinic receptors, are prevalent along its length. Working on the South American rattlesnake ( Crotalus durissus), we studied four different portions of the pulmonary artery (truncus, proximal, distal, and branches). Acetylcholine elicited robust vasoconstriction in the proximal, distal, and branch portions, but the truncus vasodilated. Electrical field stimulation (EFS) caused contractions in all segments, an effect partially blocked by atropine. We identified all five subtypes of muscarinic receptors (M1–M5). The expression of the M1 receptor was largest in the distal end and branches of ...
Environmental science and pollution research international, Jan 21, 2018
Cyanobacterial proliferation in river and lakes is the result of eutrophication. The cyanobacteri... more Cyanobacterial proliferation in river and lakes is the result of eutrophication. The cyanobacterium Radiocystis fernandoi strain R28 produces mostly two MC variants MC-RR and MC-YR and small amounts of other oligopeptides, but does not produce MC-LR. The present study investigated the hepatotoxic potential of the crude extract of the R. fernandoi strain R28 on the Neotropical fish, Piaractus mesopotamicus, at 3, 6, and 24 h after intraperitoneal injection (100 μg MC-LR equivalent per kgbody mass) using biochemical and morphological biomarkers of liver damage. Although the protein phosphatases PP1 and PP2A were not inhibited during the 24-h treatment, liver parenchyma and hepatocyte structure were disrupted. Alkaline phosphatase increased at 3 h post-injection and decreased after 24 h; alanine aminotransferase and aspartate aminotransferase increased in a time-dependent manner up to 24 h indicating impaired liver function. Progressive histopathological changes were consistent with bi...
Alternagin-C (ALT-C) is a disintegrin-like protein isolated from Rhinocerophis alternatus snake v... more Alternagin-C (ALT-C) is a disintegrin-like protein isolated from Rhinocerophis alternatus snake venom, which induces endothelial cell proliferation and angiogenesis. The aim of this study was to evaluate the systemic effects of a single dose of alternagin-C (0.5 mg·kg(-1), via intra-arterial) on oxidative stress biomarkers, histological alterations, vascular endothelial growth factor (VEGF) production, and the degree of vascularization in the liver of the freshwater fish traíra, Hoplias malabaricus, seven days after the initiation of therapy. ALT-C treatment increased VEGF levels and hepatic angiogenesis. ALT-C also enhanced hepatic antioxidant enzymes activities such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, decreasing the basal oxidative damage to lipids and proteins in the fish liver. These results indicate that ALT-C improved hepatic tissue and may play a crucial role in tissue regeneration mechanisms.
<b>Suppl. Fig. 1.</b> Detection of (A) glyceraldehyde 3-phosphate dehydrogenase (Gapd... more <b>Suppl. Fig. 1.</b> Detection of (A) glyceraldehyde 3-phosphate dehydrogenase (Gapdh), (B) clathrin (Cltc), (C) Chrm1, (D) Chrm2, (E) Chrm3, (F) Chrm4, (G) Chrm5, (H) Chat by RT-PCR of pulmonary arterial segments from the South American rattlesnake, <i>Crotalus</i> <i>durissus</i>. The products were sequenced and confirmed detection of the right gene product. (A, B) RT positive samples from 1: Proximal, 2: Distal, 3: Branches, 4: Truncus, 5: Proximal, 6: Branches of the pulmonary artery. (C, D, F, G) RT positive samples from 1: Proximal, 2: Distal, 3: Branches, 4: Truncus, 5: Proximal, 6: Branches of the pulmonary artery, while RT negative (genomic contamination) is checked for the same samples in -1 to -6. (E, H) RT positive samples from 1: Proximal, 2: Distal, 3: Branches of the pulmonary artery, while RT negative (genomic contamination) is checked for the same samples in -1 to -3. Primers can be seen in Table 1.
The South American rattlesnake, Crotalus durissus, has been successfully used as an experimental ... more The South American rattlesnake, Crotalus durissus, has been successfully used as an experimental model to study control of the cardiovascular system in squamate reptiles. Recent technical advances, including equipment miniaturization, have lessened the impact of instrumentation on in vivo recordings, and an increased range of anesthetic drugs has improved recording conditions for in situ preparations. Nevertheless, any animal-based experimental approach has to manage limitations regarding the avoidance of pain and stress the stability of the preparation and duration of experiments and the potentially overriding effects of anesthesia. To address such aspects, we tested a new experimental preparation, the decerebrate rattlesnake, in a study of the autonomic control of cardiovascular responses following the removal of general anesthesia. The preparation exhibited complex cardiovascular adjustments to deal with acute increases in venous return (caused by tail lifting), to compensate for blood flow reduction in the cephalic region (caused by head lifting), for body temperature control (triggered by an external heating source), and in response to stimulation of chemoreceptors (triggered by intravenous injection of NaCN). The decerebrate preparation retained extensive functional integrity of autonomic centers, and it was suitable for monitoring diverse cardiac and vascular variables. Furthermore, reanesthetizing the preparation markedly blunted cardiovascular performance. Isoflurane limited the maintenance of recovered cardiovascular variables in the prepared animal and reduced or abolished the observed cardiovascular reflexes. This preparation enables the recording of multiple concomitant cardiovascular variables for the study of mechanistic questions regarding the central integration of autonomic reflex responses in the absence of anesthesia.
Understanding the basis of vascular tonus regulation is fundamental to comprehending cardiovascul... more Understanding the basis of vascular tonus regulation is fundamental to comprehending cardiovascular physiology. In the present study, we used the recently developed decerebrate rattlesnake preparation to investigate the role of nitric oxide (NO) in the control of vascular tonus in a squamate reptile. This preparation allowed multiple concomitant cardiovascular parameters to be monitored, while avoiding the deleterious effect of anesthetic drugs on autonomic modulation. We observed that both systemic and pulmonary circuits were clearly responsive to NO signaling. NO increased vascular conductance in the systemic and pulmonary systems. Vasodilation by NO of the systemic circulation was compensated by cardiovascular alterations involving venous return, cardiac output, and cardiac shunt adjustments. The cardiac shunt seemed to be actively used for hemodynamic adjustments via modulation of the pulmonary artery constriction. N(ω)-nitro-L-arginine methyl ester injection demonstrated that NO contributes to modulating resting vasodilation in the systemic circuit. In contrast, NO-mediated vasodilation did not have an important role in the pulmonary circulation in inactive decerebrated snakes at 25°C. These responses vary importantly from those described for anesthetized snakes.
American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 2020
Vascular tone in the reptilian pulmonary vasculature is primarily under cholinergic, muscarinic c... more Vascular tone in the reptilian pulmonary vasculature is primarily under cholinergic, muscarinic control exerted via the vagus nerve. This control has been ascribed to a sphincter located at the arterial outflow, but we speculated whether the vascular control in the pulmonary artery is more widespread, such that responses to acetylcholine and electrical stimulation, as well as the expression of muscarinic receptors, are prevalent along its length. Working on the South American rattlesnake ( Crotalus durissus), we studied four different portions of the pulmonary artery (truncus, proximal, distal, and branches). Acetylcholine elicited robust vasoconstriction in the proximal, distal, and branch portions, but the truncus vasodilated. Electrical field stimulation (EFS) caused contractions in all segments, an effect partially blocked by atropine. We identified all five subtypes of muscarinic receptors (M1–M5). The expression of the M1 receptor was largest in the distal end and branches of ...
Environmental science and pollution research international, Jan 21, 2018
Cyanobacterial proliferation in river and lakes is the result of eutrophication. The cyanobacteri... more Cyanobacterial proliferation in river and lakes is the result of eutrophication. The cyanobacterium Radiocystis fernandoi strain R28 produces mostly two MC variants MC-RR and MC-YR and small amounts of other oligopeptides, but does not produce MC-LR. The present study investigated the hepatotoxic potential of the crude extract of the R. fernandoi strain R28 on the Neotropical fish, Piaractus mesopotamicus, at 3, 6, and 24 h after intraperitoneal injection (100 μg MC-LR equivalent per kgbody mass) using biochemical and morphological biomarkers of liver damage. Although the protein phosphatases PP1 and PP2A were not inhibited during the 24-h treatment, liver parenchyma and hepatocyte structure were disrupted. Alkaline phosphatase increased at 3 h post-injection and decreased after 24 h; alanine aminotransferase and aspartate aminotransferase increased in a time-dependent manner up to 24 h indicating impaired liver function. Progressive histopathological changes were consistent with bi...
Alternagin-C (ALT-C) is a disintegrin-like protein isolated from Rhinocerophis alternatus snake v... more Alternagin-C (ALT-C) is a disintegrin-like protein isolated from Rhinocerophis alternatus snake venom, which induces endothelial cell proliferation and angiogenesis. The aim of this study was to evaluate the systemic effects of a single dose of alternagin-C (0.5 mg·kg(-1), via intra-arterial) on oxidative stress biomarkers, histological alterations, vascular endothelial growth factor (VEGF) production, and the degree of vascularization in the liver of the freshwater fish traíra, Hoplias malabaricus, seven days after the initiation of therapy. ALT-C treatment increased VEGF levels and hepatic angiogenesis. ALT-C also enhanced hepatic antioxidant enzymes activities such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, decreasing the basal oxidative damage to lipids and proteins in the fish liver. These results indicate that ALT-C improved hepatic tissue and may play a crucial role in tissue regeneration mechanisms.
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