Effect of histological processing and methacrylate sectioning on the area of gill tissue in teleost.

Deformation of biological tissues may occur during histological processing and results in loss of accuracy when quantitative information about cells, tissues and organs is necessary. In this study, the gill tissue from armored catfish (Pterygoplichthys anisitsi) was quantified in each step of processing using the stereological principles. During processing for glycol methacrylate embedding, gill tissue from shrinks significantly but regains its original dimensions after sectioning.

Effect of histological processing and methacrylate sectioning on the area of gill tissue in teleost / Efeito do processamento histológico e seccionamento em metacrilato sobre a área de tecido branquial em teleósteo

Deformation of biological tissues may occur during histological processing and results in loss of accuracy when quantitative information about cells, tissues and organs is necessary. In this study, the gill tissue from armored catfish (Pterygoplichthys anisitsi) was quantified in each step of processing using the stereological principles. During processing for glycol methacrylate embedding, gill tissue from shrinks significantly but regains its original dimensions after sectioning.

Deformações nos tecidos podem ocorrer durante o processamento histológico e resultar em informações errôneas quando há necessidade de dados quantitativos sobre células, tecidos e órgãos. Neste estudo, o tecido branquial do cascudo (Pterygoplichthys anisitsi) foi quantificado em cada etapa do processamento utilizando os princípios de estereologia. O tecido branquial reduziu significativamente durante processamento histológico com metacrilato, mas retornou às suas dimensões iniciais depois de seccionadas, o que indica não ocorrer nenhuma perda na informação quantitativa do tecido.

Pulmonary oxygen diffusing capacity of the South American lungfish Lepidosiren paradoxa: physiological values by the Bohr method.

Lungfish (Dipnoi) may represent the sister group to all land vertebrates and are therefore important for reconstructing the conquest of land by tetrapods. We determined venous and arterial blood gases, pulmonary O(2) uptake, and the form of the hemoglobin-O(2) dissociation curves in the South American lungfish Lepidosiren paradoxa. Measurements were performed at 25 degrees and 35 degrees C. Based on this information, we calculated its pulmonary O(2) diffusing capacity (D(L)O(2)), using the Bohr integration procedure. D(L)O(2) increased with temperature to reach about 0.04 mL stpd kg(-1) min(-1) mmHg(-1) at 35 degrees C. This value represents about 40% of the morphometric diffusing capacity and is similar to physiological values in some amphibians and reptiles.

Cardiorespiratory responses to hypercarbia in tambaqui Colossoma macropomum: chemoreceptor orientation and specificity.

Experiments were carried out to test the hypothesis that ventilatory and cardiovascular responses to hypercarbia (elevated water P(CO2)) in the tambaqui Colossoma macropomum are stimulated by externally oriented receptors that are sensitive to water CO(2) tension as opposed to water pH. Cardiorespiratory responses to acute hypercarbia were evaluated in both the absence and presence of internal hypercarbia (elevated blood P(CO2)), achieved by treating fish with the carbonic anhydrase inhibitor acetazolamide. Exposure to acute hypercarbia (15 min at each level, final water CO(2) tensions of 7.2, 15.5 and 26.3 mmHg) elicited significant increases in ventilation frequency (at 26.3 mmHg, a 42% increase over the normocarbic value) and amplitude (128%), together with a fall in heart rate (35%) and an increase in cardiac stroke volume (62%). Rapid washout of CO(2) from the water reversed these effects, and the timing of the changes in cardiorespiratory variables corresponded more closely to the fall in water P(CO2) (Pw(CO2)) than to that in blood P(CO2) (Pa(CO2)). Similar responses to acute hypercarbia (15 min, final Pw(CO2) of 13.6 mmHg) were observed in acetazolamide-treated (30 mg kg(-1)) tambaqui. Acetazolamide treatment itself, however, increased Pa(CO2) (from 4.81+/-0.58 to 13.83+/-0.91 mmHg, mean +/-S.E.M.; N=8) in the absence of significant change in ventilation, heart rate or cardiac stroke volume. The lack of response to changes in blood P(CO2) and/or pH were confirmed by comparing responses to the bolus injection of hypercarbic saline (5% or 10% CO(2); 2 ml kg(-1)) into the caudal vein with those to the injection of CO(2)-enriched water (1%, 3%, 5% or 10% CO(2); 50 ml kg(-1)) into the buccal cavity. Whereas injections of hypercarbic saline were ineffective in eliciting cardiorespiratory responses, changes in ventilation and cardiovascular parameters accompanied injection of CO(2)-laden water into the mouth. Similar injections of CO(2)-free water acidified to the corresponding pH of the hypercarbic water (pH 6.3, 5.6, 5.3 or 4.9, respectively) generally did not stimulate cardiorespiratory responses. These results are in agreement with the hypothesis that in tambaqui, externally oriented chemoreceptors that are predominantly activated by increases in water P(CO2), rather than by accompanying decreases in water pH, are linked to the initiation of cardiorespiratory responses to hypercarbia.

The frog brainstem preparation as a model for studying the central control of breathing in tetrapods

Our frog brainstem preparation revealed mechanisms for the central control of breathing that are in many ways similar to those of mammals. Thus, the basic control mechanisms for air-breathing appear to have been present in the Devonian common ancestors of frogs and mammals and may be common to all lung-breathing vertebrates. Location: The in vitro frog brainstem, including motor nuclei of cranial nerves V to X, maintains frequency and ratio of fictive buccal oscillations to fictive lung inflation episodes comparable with that of the living animal. In this preparation, transaction caudal to V abolishes spontaneous discharge in X but slow, spontaneous discharge in V may remain. Independent central pattern generation is present in the left and right half-brainstems. Chemosensitivity: The frequency of fictive lung inflation increases with decrease in pH within the physiological range. Response to glutamate: Biphasic response, consisting of a pause, followed by a dramatic increase in the frequency of fictive inspirations and positive baseline deflection, followed, in turn, by slow return of the baseline to the control level with frequency remaining above control as long as glutamate is applied. Local application reveals glutamate-sensitive sites in the ventral reticular formation. Response to substance P and physalaemin: Similar to glutamate but the frequency of fictive inspirations decreases below control values. Response to strychnine: The normal temporal sequence in firing of motor neurons of cranial nerves is disrupted and all nerves are synchronously active. The firing sequence of respiratory neurons is consistent with a grouping possibly homologous to the mammalian inspiratory, post-inspiratory and expiratory phases.

The frog brainstem preparation as a model for studying the central control of breathing in tetrapods.

Our frog brainstem preparation revealed mechanisms for the central control of breathing that are in many ways similar to those of mammals. Thus, the basic control mechanisms for air-breathing appear to have been present in the Devonian common ancestors of frogs and mammals and may be common to all lung-breathing vertebrates. LOCATION: The in vitro frog brainstem, including motor nuclei of cranial nerves V to X, maintains frequency and ratio of fictive buccal oscillations to fictive lung inflation episodes comparable with that of the living animal. In this preparation, transection caudal to V abolishes spontaneous discharge in X but slow, spontaneous discharge in V may remain. Independent central pattern generation is present in the left and right half-brainstems. CHEMOSENSITIVITY: The frequency of fictive lung inflations increases with decrease in pH within the physiological range. RESPONSE TO GLUTAMATE: Biphasic response, consisting of a pause, followed by a dramatic increase in the frequency of fictive inspirations and positive baseline deflection, followed, in turn, by slow return of the baseline to the control level with frequency remaining above control as long as glutamate is applied. Local application reveals glutamate-sensitive sites in the ventral reticular formation. RESPONSE TO SUBSTANCE P AND PHYSALAEMIN: Similar to glutamate but the frequency of fictive inspirations decreases below control values. RESPONSE TO STRYCHNINE: The normal temporal sequence in firing of motor neurons of cranial nerves is disrupted and all nerves are synchronously active. The firing sequence of respiratory neurons is consistent with a grouping possibly homologous to the mammalian inspiratory, post-inspiratory and expiratory phases.