RESUMO
Perilymph and serum of guinea pigs were investigated using immunoelectrophoresis and polyacrylamide gel electrophoresis. The molecular weights of the perilymph proteins were estimated by electrophoresis in a linear polyacrylamide gel gradient (3-20%). The immunoelectropherograms and the polyacrylamide gel electropherograms of equivalent amounts of perilymph protein and serum protein are nearly the same. The immunological detection of the perilymph proteins indicates that they are serum proteins. The clear identification of the individual proteins fails because of missing monospecific antisera against the individual proteins of guinea pig. High-molecular proteins (range of the molecular weight estimation 67 000-290 000) are also detectable in the perilymph according to the molecular weight estimation.
Assuntos
Líquidos Labirínticos/análise , Perilinfa/análise , Proteínas/análise , Animais , Proteínas Sanguíneas/análise , Eletroforese em Gel de Poliacrilamida , Cobaias , Imunoeletroforese , Peso MolecularRESUMO
The protein concentration of the guinea pig perilymph was investigated systematically using a micro-modification of the method of Lowry et al. Perilymph of scala vestibuli and of scala tympani was obtained from living animals and immediately post mortem by various methods. In living animals it is especially difficult to obtain samples without blood contamination. Another problem in the obtaining of perilymph from living animals is the contamination of tympanic perilymph samples with cerebrospinal fluid. This contamination diminishes the protein concentration of perilymph to a high degree. When the subarachnoid space is opened suboccipitally before perilymph extraction, there is no significant difference between protein content in tympanic and vestibular perilymph. The mean protein concentration in both cochlea scales is about 150 mg/100 ml. When samples are extracted post mortem from animals perfused intra-arterially, mean values of protein are in the same range. Without perfusion of animals, the mean value of tympanic samples extracted post mortem is significantly higher. Causes of artefacts in perilymph investigations are discussed.
Assuntos
Líquidos Labirínticos/metabolismo , Perilinfa/metabolismo , Proteínas/metabolismo , Animais , Cóclea/metabolismo , Cobaias , Perfusão , Perilinfa/análise , Proteínas/análise , Espaço Subaracnóideo/metabolismo , Membrana Timpânica/metabolismoRESUMO
The protein concentration in the perilymph of noise-exposed guinea pigs was investigated using a micromethod described previously. The animals were exposed to wide band noise at 140 dB for one hour unilaterally in a closed acoustic system. Perilymph was obtained after exposure at different intervals: immediately, 6 or 24 hours later. From animals exposed to noise, it is particularly more difficult to obtain perilymph without blood contamination than from normal ones. Even post mortem, it is difficult to obtain samples without blood contamination and without hemolysis, particularly from the scala tympani. Therefore, some animals were perfused intra-arterially with Ringer solution or with Infukoll M 40 before decapitation. After exposure to noise only a small increase in protein concentration of the perilymph can be detected in the noise-exposed ears of animals perfused intra-arterially. The difference in concentration is most distinct in samples extracted 6 hours after noise exposure had ceased. No reliable clue to the source of this protein could be obtained from these investigations.
Assuntos
Líquidos Labirínticos/análise , Ruído , Perilinfa/análise , Proteínas/análise , Animais , Membrana Basilar/fisiologia , Feminino , Cobaias , Perda Auditiva Provocada por Ruído/metabolismo , Fatores de Tempo , Membrana Timpânica/fisiologia , Vestíbulo do Labirinto/fisiologiaRESUMO
The paper deals with comparative studies of lactate concentration in the perilymph (PL) of scala tympani and of scala vestibuli, arterial and venous blood, serum and cerebrospinal fluid (CSF) of normal and sound exposed guinea pigs, special consideration having been given to possible sources of error in the methods employed. Lactate was determined enzymatically using a micromodification of the Boehringer UV-test combination adapted to 1 mul PL. The lactate concentrations in the PL of scala tympani and scala vestibuli did not differ significantly. The mean values amounted to 4.5-5.2 mM/l in the case of the opened and of the unopened subarachnoid space (Table 1). The lactate concentrations in the PL of both cochlea scales were significantly higher already ten minutes post-mortem. In the exposure experiments the animals were unilaterally exposed to sound for 1 h in an acoustically isolated system using a wide-band noise at an intensity of 120 dB SPL for one series and 2-kHz pure-tone at intensities of 112 and 122 dB SPL for two other series. We did not detect any changes in the lactate concentrations neither in the PL nor in the blood and in the CSF, following sound exposure (Table 2 and 3). The lactate concentrations of arterial and venous blood and CSF did not differ significantly. The mean values amounted to 1.4-1.8 mM/l (Table 2). However, if blood was not deproteinized or centrifuged immediately after being taken, the lactate concentration increased markedly. A comparison of the present results has shown that the lactate concentration in the PL is about three times as high as in blood and in CSF. This difference in concentration suggests that the PL lactate is of intracochlear origin and that glycolytic processes take place in the inner ear also under normal conditions. Systematic studies of additional metabolic parameters must be conducted before a definitive physiological interpretation of the present analytical results can be given.
Assuntos
Líquidos Labirínticos/metabolismo , Lactatos/metabolismo , Perilinfa/metabolismo , Som , Animais , Cobaias , Lactatos/sangue , Lactatos/líquido cefalorraquidianoRESUMO
Lactate and pyruvate of perilymph (PL) were studied 30, 60, and 120 min postmortem. During this period the mean lactate concentration of scala tympani and scala vestibuli increased from 4.8 mmol/l found intravitally to 17.8 and 15.1 mmol/l, respectively, whereas pyruvate decreased from an average of 0.33 to 0.10 mmol/l (fig. 1). These inverse changes of concentration yield postmortem lactate/pyruvate quotients which are more than one order of magnitude higher than the quotients found intravitally (Table 1). In comparative tests of blood samples carried out 30, 60, and 120 min after the sampling (Fig.1), the lactate increase was found to be markedly lower than in postmortem PL. The substantial metabolite changes in PL seem to be caused by glycolytic activity of all cochlear structures that are in direct contact with PL. The decrease of pyruvate level is probably due to a shift of the lactate-pyruvate equilibrium (lactate dehydrogenase system) in PL. The blood vessels in the perilymphatic space can be neglected as postmortem metabolite source of PL.
Assuntos
Líquidos Labirínticos/metabolismo , Lactatos/metabolismo , Perilinfa/metabolismo , Mudanças Depois da Morte , Piruvatos/metabolismo , Animais , Cobaias , Ácido Láctico , Ácido Pirúvico , Rampa do Tímpano/metabolismo , Vestíbulo do Labirinto/metabolismoRESUMO
Lactate and pyruvate were studied comparatively in perilymph (PL), blood, and cerebrospinal fluid (CSF) of anesthetized guinea pigs. Arterial blood pressure, heart and respiration rate (Fig. 1), and arterial blood-gas state (Table 2) were simultaneously checked in a group of the animals. The metabolites were determined enzymatically by using the fluorometric technique. The studies have shown (Table 1) that both the lactate and the pyruvate concentrations are in PL at a similar rate (about 3:1) higher than in native blood and also higher than in CSF. The metabolite values of blood, especially the lactate values, were lower when blood was taken alone, e.g., more physiological, than in the case when CSF and PL had been sampled before. The lactate/pyruvate ratios of Pl are somewhat higher than the blood ratios. The ratio of CSF was found to be lower. The high metabolite levels in PL suggest an intracochlear origin. A direct perilymphatic lactate origin could not be detected.
Assuntos
Líquidos Labirínticos/análise , Lactatos/análise , Perilinfa/análise , Piruvatos/análise , Animais , Gasometria , Cobaias , Humanos , Lactatos/sangue , Lactatos/líquido cefalorraquidiano , Piruvatos/sangue , Piruvatos/líquido cefalorraquidianoRESUMO
Glucose, pyruvate, and lactate of perilymph (PL), blood, and cerebrospinal fluid (CSF) of unexposed and sound-exposed guinea pigs under ethyl urethane anesthesia were examined with due consideration of the principal sources of error. The animals had fasted for 15--20 h before the experiment to stabilize the blood glucose level. The metabolites were determined enzymatically by means of fluorescence measurements. It was found that the glucose levels depend not only on ingestion but also on the duration of anesthesia of the animals before sampling. The mean values of the scala tympani and scala vestibuli PL and CSF did not differ significantly, being about half those of blood or plasma immediately (10--20 min) after introducing anesthesia (Table 2). This concentration difference is in disagreement with the original ultrafiltration hypothesis of PL, suggesting a blood-PL barrier for glucose. The dependence on the duration of anesthesia and on the animals' ingestion before sampling appears to be an important cause of the differences in glucose data published in literature hitherto. No influence of anesthesia on pyruvate and lactate concentrations was observed. Data obtained on unexposed control animals (Tables 3 and 4) confirmed our earlier metabolite findings (Scheibe et al. 1976, 1981). No major changes in glucose, pyruvate, and lactate concentration of PL, blood, and CSF were detectable immediately after 1 h of exposure to wide-band noise at an intensity of 120 dB SPL. The present lactate findings confirmed our earlier exposure experiments (Scheibe et al. 1976), but they did not agree with the information given by Schnieder (1974).
Assuntos
Glicemia/metabolismo , Perda Auditiva Provocada por Ruído/metabolismo , Lactatos/metabolismo , Piruvatos/metabolismo , Animais , Cobaias , Perilinfa/metabolismo , Rampa do Tímpano/metabolismo , Vestíbulo do Labirinto/metabolismoRESUMO
The paper deals with: 1. the protein concentration in the perilymph (PL), the serum and the cerebrospinal fluid (CSF), 2. the protein pattern in the PL and 3. histological findings in the middle and inner ear in unilaterally ear-infected guinea pigs. The studies were performed 6 h to 21 days post infectionem (Fig. 1). The pathological changes in the middle ear, which, in most cases, were limited to the infected ear, were initially evaluated under the operating microscope and divided into 4 stages. The analytical and histological results were presented as functions of these stages. As the inflammation intensity increased, the protein concentration in the PL of the infected ears increased to a level exceeding that of the normal value more than ten times (Fig. 2). However, in the serum and in the CSF this concentration remained unchanged. Likewise, no significant protein increase in the PL of the contralateral ears was detectable in most cases. As the inflammation intensity increased, the number of the precipitation lines detectable immunoelectrophoretically increased in the PL of the infected ears (Fig. 3). An increase in the alpha1- and gamma-globulins and a decrease in Albumin was found by electrophoresis on cellulose acetate strips (Tab. 3). The histological findings correlated with initially established inflammatory stages of the middle ear mucous membrane (Tab. 4). As the inflammation intensity increased, the round window, too, was changed pathologically, so that in some cases of purulent otitis media middle ear secretion could enter the cochlea. The protein increase in the PL immediately after the infection is probably due to an increase in the blood vessel permeability in the inner ear.