Early postnatal sound exposure induces lasting neuronal changes in the inferior colliculus of senescence accelerated mice (SAMP8): a morphometric study on GABAergic neurons and NMDA expression

Senescence-acceleration-prone mice (SAMP8) provide a model to study the influence of early postnatal sound exposure upon the aging auditory midbrain. SAMP8 were exposed to a 9-kHz monotone of either 53- or 65-dB sound pressure level during the first 30 postnatal days, the neurons in the auditory midbrain responding selectively to 9 kHz were localized by c-fos immunohistochemistry and the following parameters were compared to control SAMP8 not exposed to sound: mortality after sound exposure, dendritic spine density, and quantitative neurochemical alterations in this 9-kHz isofrequency lamina. For morphometric analysis, animals were examined at 1, 4, and 8 months of age. Serial sections of the inferior colliculus were Golgi impregnated or stained immunohistochemically for the expression of 1 subunit of NMDA receptor or GABA. Mortality after exposure to 53 dB was the same as in controls, but was markedly increased from 7 months of age onward after postnatal exposure to 65 dB. No gross morphological alterations were observed in the auditory midbrain after sound exposure. However, sound exposure to 53 or 65 dB significantly reduced dendritic spine density by 11% at 4 months or by 11–17% both at 1 and 4 months of age, respectively. The effect of sound exposure upon neurons expressing the NMDA1 subunit was dose-dependent. Increasing with age until 4 months in control mice and remaining essentially stable thereafter, the percentage of NMDA1-immunoreactive neurons was significantly elevated by 40–66% in 1- and 8-month-old SAMP8 exposed to 53 dB, whereas no significant effect of 65 dB was apparent. The proportion of GABAergic cells declined with age in controls. It was significantly decreased at 1 month after 53 and 65 dB sound exposure. In contrast, it was elevated at later stages, being significantly increased at 4 months after exposure to 53 dB and at 8 months after exposure to 65 dB. The total cell number in the 9-kHz isofrequency lamina of SAMP8 decreased with age, but was not affected by exposure to either 53 or 65 dB. The present results indicate that early postnatal exposure to a monotone of mild intensity has long-term effects upon the aging auditory brain stem. Some of the changes induced by sound exposure, e.g., decline in spine density, are interpreted as accelerations of the normal aging process, whereas other effects, e.g., increased NMDA1 expression after 53 dB and elevated GABA expression after both 53 and 65 dB, are not merely explicable by accelerated aging.     

Calling song recognition in the cricket Teleogryllus oceanicus: comparison of the effects of stimulus intensity and sound spectrum on selectivity for temporal pattern

The turning response to surface waves of clawed toads (Xenopus laevis) with an inactivated lateral line was reinvestigated to examine whether sensory systems other than the lateral line (second systems) are involved. Two methods were used to block the lateral line input: selective and reversible inactivation of the lateral line periphery using CoCl₂ or chronic destruction with thermocautery. The time-course of the response recovery (response frequency, turning accuracy and reaction time) was recorded. Following CoCl₂ inactivation 10 out of 13 animals did not respond to surface waves for at least 2 days. The remaining 3 animals gave sporadic turning responses. It is assumed that in these individuals a second system is permanently involved in the detection of surface waves parallel to the lateral line. Five days after the chronic destruction of the lateral line all animals again turned to the centre of surface waves. It is suggested that by this time the second system had become capable of substituting for the missing lateral line input. The response frequency and the accuracy of the turning response of lesioned animals varied considerably among individuals but was always lower than in untreated animals (tested up to 120 days).Abbreviation LED light emitting diode     

Observation of Brillouin scattering from single muscle fibres

The propagation of sound waves along relaxed single fibres of glycerinated rabbit psoas muscle has been observed using Brillouin scattering at frequencies up to 1.6 GHz. Two types of waves were observed: one with a velocity of 1508±7 m s^–1, which is attributed to sound waves in intra-cellular saline, the other with a velocity of 912±25 m s^–1, which is attributed to waves propagating along the protein filaments within individual sarcomeres. The latter sound velocity is much higher than that which has been reported by Stienen and Blangé (1985) for 50 s tension transients, and the difference is attributed to the much higher stiffness of the protein filaments compared to the cross-bridges which determine the low-frequency elasticity of muscle fibres.     

Sound waves increases the ascorbic acid content of alfalfa sprouts by affecting the expression of ascorbic acid biosynthesis-related genes

Previous studies have shown that sound wave treatment can affect the expression of plant genes and improve the growth. So, we investigated the ability of sound waves to increase AsA (l-ascorbic acid) content in alfalfa (Medicago sativa) sprouts in this study. Sprouts were exposed to a range of sound wave frequencies for two 1-h periods per day for various numbers of days. Most sound wave treated sprouts had a higher AsA content than untreated sprouts. In addition, the activity level of superoxide dismutase, an enzyme with potent antioxidative properties, was increased in sound wave-treated sprouts. The AsA content varied in response to sound wave treatment. Most processing conditions, including 500 and 1000 Hz, increased AsA content by 24–50%; however, some treatment conditions caused reduced AsA content during sprout growth. Furthermore, AsA content during sprout storage was increased by most sound wave treatment conditions, with 13–36% increases observed following 800 and 1000 Hz sound wave treatments compared to untreated sprouts. To investigate the mechanisms underlying changes in AsA content, we analyzed the expression levels of AsA biosynthesis-related genes. We found that several genes, including VTC1, VTC2, VTC4, GME, L-GalDH, GLDH, MDHAR, and DHAR1, displayed differential expression in response to sound wave treatment. Therefore, sound wave treatment may be a viable method for increasing the nutritional contents of sprouted vegetables.     

Effect of chronic microwave radiation on T cell-mediated immunity in the rabbit

Experiments were conducted to elucidate the effects of chronic low power-level microwave radiation on the immunological systems of rabbits. Fourteen male Belgian white rabbits were exposed to microwave radiation at 5 mW/cm², 2.1 GHz, 3 h daily, 6 days/week for 3 months in two batches of 7 each in specially designed miniature anechoicchambers. Seven rabbits were subjected to sham exposure for identical duration. The microwave energy was provided through S band standard gain horns connected to a 4K3SJ2 Klystron power amplifier. The first batch of animals were assessed for T lymphocyte-mediated cellular immune response mechanisms and the second batch of animals for B lymphocyte-mediated humoral immune response mechanisms. The peripheral blood samples collected monthly during microwave/sham exposure and during follow-up (5/14 days after termination of exposures, in the second batch animals only) were analysed for T lymphocyte numbers and their mitogen responsiveness to ConA and PHA. Significant suppression of T lymphocyte numbers was noted in the microwave group at 2 months (P<0.01, % 21.5%) and during follow-up (P<0.01, % 30.2%). The first batch animals were initially sensitised with BCG and challenged with tuberculin (0.03 ml) at the termination of microwave irradiation/sham exposure and the increase in foot pad thickness ( mm), which is a measure of T cell-mediated immunity (delayed type hypersensitivity response, DTH) was noted in both the groups. The microwave group revealed a better response than the control group (%+12.4 vs.+7.54). The animals were sacrified and the tissue T lymphocyte counts (spleen and lymph node) were analysed. No significant variation was observed in the tissue T lymphocyte counts of microwave-irradiated rabbits. From these results it is speculated that the T lymphocytes are sequestered to various lymphoid organs under the influence of microwaves. A sub-population of T cells known as T helper cells (mediating DTH response) are probably not affected by microwave radiation. It is clear from our experiments that although chronic microwave radiation at 5 mW/cm² leads to suppression of peripheral T lymphocyte numbers, there is no concomitant functional impairment of these cells as evidenced by functional assays.     

Sterilizing Effects of High-Intensity Airborne Sonic and Ultrasonic Waves

The lethal effects of high-intensity airborne sonic (9.9 kc/sec) and ultrasonic waves (30.4 kc/sec) on spores of Bacillus subtilis var. niger ATCC 9372 were determined. The spores, which were deposited on filter-paper strips, were exposed to sound waves for periods varying from 1 to 8 hr, at a temperature of 40 C and a relative humidity of 40%. Significant reductions in the viable counts of spores exposed to airborne sonic or ultrasonic irradiations were obtained. The antibacterial activity of airborne sound waves varied with the sound intensity level, the period of irradiation, and the distance of the sample from the sound source. At similar intensity levels, the amplitude of motion of the sound waves appeared to be a factor in acoustic sterilization.     

High blood lead level among garage workers in Bangkok, public concern is necessary

Lead is an important toxic metal agent found in many industrial processes in the present day. Lead exposure must be of particular concern because of ongoing exposure to thousands of workers in the industrial plants and recent research indicating that asymptomatic lead exposure can result in chronic toxicity manifestations. Therefore, determination and control lead exposure among the risk workers is very necessary. Like other developing countries, lead pollution becomes an important public health problem of Thailand, especially for the big cities as Bangkok but relatively few of these countries have introduced policies and regulations for significantly combating the problem. We set this pilot study to determine the blood lead levels by anodic stripping volammetry (ASV) method as a marker for lead exposure among the occupational exposed and control subjects. Totally 89 subjects, 20 control subjects and 69 garage workers (52 mechanics and 17 dye sprayers), as the representatives of occupational exposed subjects, were included into this preliminary study. The mean blood lead level in the control group was 0.32±0.07 mol/l. The mean blood lead level in the mechanics group was 0.42±0.13 mol/l. The mean blood lead level in the dye sprayers was 0.58±0.07 mol/l. Significant higher blood lead levels among the mechanics and dye sprayer groups were observed (P<0.05). Based on this study, the considerations for prevention of possibly exposure to lead among the high-risk workers as public health policies was recommended.     

Alterations in enzymes involved in fat metabolism after acute and chronic altitude exposure

The purpose of this study was toexamine the effect of acute (24 h) and chronic (5 wk) hypobaric hypoxicexposure equivalent to a simulated altitude of 4,300 m (446 mmHg) onthe enzymes of fat metabolism. Heart, liver, and skeletal muscle weretaken from 32 male Sprague-Dawley rats. Altitude exposure did notaffect the activity of citrate synthase in any of the tissues,suggesting that mitochondrial content was unchanged. Carnitinepalmitoyltransferase-I (CPT-I) activity was significantly reduced inthe heart by both acute and chronic high altitude exposure comparedwith controls. A similar reduction was found for CPT-I activity inextensor digitorum longus after acute and chronic exposurecompared with control animals. CPT-I activity was not affected byaltitude exposure in the soleus muscle or the liver. 3-Hydroxyacyl-CoAdehydrogenase (-HAD) activity was significantly depressed in thehearts of chronically exposed animals compared with controls. Nodifference between acute and control animals was found in the heart for-HAD activity. Liver -HAD activity was also significantlydecreased in the acclimatized as well as in the acute animals comparedwith the control group. Quadriceps -HAD activity was reduced for the chronic animals only compared with controls. These data suggest thatacclimatization to high altitude selectively decreases key enzymes infat utilization and oxidation in the heart, liver, and select skeletal muscles.

     

Alteration of life span of mice chronically exposed to 2.45 GHz CW microwaves

Female CD 1 mice were exposed from the thirty-fifth day of age for the remainder of their lives to 2.45 GHz, CW-microwave radiation at a power density of 3 or 10 m W/cm² (SAR = 2.0 or 6.8 W/kg). Exposures took place 1 h/day, 5 day/week in an anechoic chamber at an ambient temperature of 22 °C and a relative humidity of 50%. There were 25 animals in each exposure group, and an equal number of controls were concurrently sham exposed. The average life span of animals exposed at 10 mW/cm² was significantly shorter than that of sham-exposed controls (572 days vs. 706 days; P = .049; truncation >20%). In contrast, the average lifespan of the animals exposed at 3 mW/cm² was slightly, but not significantly, longer (738 days) than that of controls (706 days). © 1994 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Analysis of surface wave direction by the lateral line system of Xenopus: Source localization before and after inactivation of different parts of the lateral line

The turning responses of clawed toads (Xenopus laevis) to surface waves were examined in animals with an intact lateral line or with different combinations of lateral lines reversibly inactivated by CoCl₂. The responses were characterized with respect to response frequency, turning accuracy, turning side, response time, and swim distance. After the inactivation most animals still responded to surface waves but the responses were different from those of animals with an intact lateral line. They also differed according to the combination of inactivated lines. In all experiments the responses for stimuli in some sectors of the surface did not differ from controls. The location of these sectors co-varied with the position of the intact lines, i.e., normal responses were found for frontal stimulus directions when head lines were intact and for caudolateral stimulus directions when trunk lines were intact. Their size was larger when lines on both sides of the body were intact and smaller when only lines on one side were intact. When the number of functional lines was reduced to one or two on one side of the body the turning angles shown within the sector of normal responses were maintained for stimulus directions outside these sectors. These results can be interpreted as indicating that head and trunk lines represent different position values. When only a single line was functional the toads still turned towards the stimulus source more often than by chance.It is hypothesized that Xenopus uses two mechanisms to determine the direction of surface waves. One uses the position values of head and trunk lines; this mechanism is comparable to the place value postulated for individual head neuromasts of surface feeding fish. The other uses the information encoded in the activity pattern that is elicited in one line when the surface wave travels over the line. This second mechanism yields information about stimulus side but not about stimulus angle.     

Neurophysiologial Study of the Hemispheric Organization of the EEG Spectrum in Mental Infantilism in Adolescents

Sixteen adolescents between 15 to 17 years of age with mental infantilism syndrome participated in the study. The data were compared with the control group including 10 mentally and neurologically healthy young subjects, secondary-school students in the 10th or 11th grades. The EEG was recorded monopolarly from 12 leads in the states of quiet wakefulness (closed eyes), on general activation (open eyes), when performing cognitive hemispheric-specialized tests, and when exposed to aversive sound stimulation. The studies showed that EEG spectral parameters could be regarded as the objective criteria of mental infantilism. In the baseline, the patients demonstrated the EEG characteristics typical of the earlier ontogenesis stage, i.e., a generalized increase in the spectral powers of -, ₁-, ₂-, and ₁-ranges and a decrease in the interhemispheric coherence in the frontal temporal region. When performing cognitive hemispheric-specialized tests, infantile patients were characterized by the insufficient differentiation of reactions to functional loads in terms of the frequency ranges, topography, and lateralization of changes, with the predominance of shifts in the ₂-range that is directly connected with nonspecific aspects of activation. On the presentation of sound stimulation, only the patients' group demonstrated an increase in the power of slow EEG ranges and interhemispheric coherence in the frontal temporal area. The results demonstrate the delayed formation of hemispheric specialization with a more retarded maturation of the nervous networks of the left hemisphere and the dysfunction of the frontal temporal regions of the brain.     

Changes of body fluid and hematology in toad and their rehabilitation following intermittent exposure to simulated high altitude

Three groups of adult male toads were exposed intermittently in a decompression chamber for a daily period of 4 and 8 hours at a time for 6 consecutive days to an altitude of 12,000; 18,000 and 24,000 feet (3658; 5486; 7315 m) respectively. Most of the exposed animals were sacrificed immediately after the last exposure, but only a few animals experiencing 8 hours of exposure were sacrificed after a further 16 hours of exposure at normal atmospheric pressure. Eight hours of daily exposure for 6 days causes a decrease of body fluids and an increase of hematological parameters in all the altitude exposed animals compared with to the changes noted in the animals having 4 hours of daily exposure for 6 days at the same altitude levels. The animals that were exposed to pressures equivalent to altitudes of 12,000 and 18,000 feet daily for 8 hours were found to return nearly to their normal body fluids and hematological balance after 16 hours of exposure to normal atmospheric pressure, whereas the animals exposed for a similar period at an equivalent 24,000 feet failed to get back their normal balance of body fluids and hematology after 16 hours of exposure at normal atmospheric pressure. The present experiment shows that the body weight loss and changes of body fluid and hematological parameters in the toad after exposure to simulated high altitude are due not only to dehydration, but suggest that hypoxia may also have a role.     

The effects of short pulses of light at night on numbers of pineal “synaptic” ribbons and serotonin N-acetyltransferase activity in male Sprague-Dawley rats

Summary To characterize further the functionally enigmatic synaptic ribbons (SR) of the mammalian pineal gland and to study possible relationships to melatonin synthesis, in the present investigation rats were exposed to short pulses of light at night when both SR numbers and serotonin N-acetyltransferase (NAT) activity are high in comparison to day-time values. Male Sprague-Dawley rats were killed at 13:00 and 01:00 h, respectively, and at 01:10 and 02:00 h after exposure to light for 10 and 60 min, respectively. The pineals were rapidly taken out and cut sagittally in half. One half was processed for electron-microscopic quantitation of SR numbers and the other half for NAT determinations. It was found that both SR numbers and NAT activity decreased significantly when the animals were exposed to light at night. Although both parameters showed corresponding changes, there was no clear-cut correlation between SR numbers and NAT activity in individual animals within a group, except after exposure to light for 60 min when a positive correlation (R = 0.939; p < 0.05) existed. After exposure to light the electron-lucent vesicles of the SR decreased in number, but the length of the SR was unchanged. These results show that numbers of pineal SR can be easily and quickly manipulated and that the presently used model may be ideal in studying the poorly understood mode in which degradation of SR occurs.Recipient of a DAAD stipend, on leave from Department of Zoology, University of Burdwan, Burdwan, India     

Der Serotoninstoffwechsel in einzelnen Hirnteilen vom Tupaia (Tupaia belangeri) bei soziopsychischem Stress

Zusammenfassung Bei sozialem Stress nehmen die Konzentrationen von Serotonin (5-HT) und 5-Hydroxyindolessigsäure (5-HIAA) in den einzelnen Hirnteilen von Tupaias zu. Diese Zunahme läßt sich durch eine erhöhte Stoffwechselgeschwindigkeit (turnover) erklären.Die Änderungen in der Stoffwechselgeschwindigkeit der einzelnen Hirnteile hängen von der Art der sozialen Belastung ab.Schon Kampf mit Unterwerfung (zweimal täglich) führt zu einem schnelleren Stoffwechsel des 5-HT in allen Hirnteilen. Belastet man die Tiere außer durch Kampf noch dadurch, daß sie das Tier, das sie besiegt hat, zwischen den Kampfpausen sehen können, so wird der Stoffwechsel des 5-HT in einzelnen Hirnteilen (Septum, Frontaler Cortex) nochmals erhöht.Eine mögliche funktioneile Bedeutung dieser Veränderungen wurde diskutiert.

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Serotonin metabolism in discrete brain areas and sociopsychic stress in tupaia (Tupaia belangeri)

Summary The physiological state of the animals depends not only on ecological factors. It is also determined by the social interactions of the animals. Some of these social interactions are harmful to the physiological state of the individuals. The aim of this work was to separate the physical loads of social interactions from the mere psychic ones and to correlate them with neurochemical changes. For this purpose the concentration levels of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were measured in discrete brain areas.To separate the psychic from the physical loads male tupaias (Tupaia belangeri) were exposed two times a day (at 9.30 and 13.30 h) for two minutes to the attacks of an experienced fighter. The animals were defeated during these attacks. One group of the defeated animals was in optical contact with the experienced fighter throughout the day (12 hours; Stresstiere). The other group of the defeated animals was optically separated from the experienced fighter after each fight (Kampftiere).After four days the animals were decapitated at 15.00 h, 5.5 hours after the last fight (on the fourth day the animals were not allowed to fight at 13.30 h). Animals which had not fought were decapitated at the same time and served as controls. 5-HT and 5-HIAA levels were estimated within two days after decapitation.During the test the animals lose body weight. Animals who are only exposed to attacks (Kampftiere) loose 2.8±2.8%; animals who are exposed to attacks and optical contact with the experienced fighter (Stresstiere) lose 11.8±3.4%.The concentration levels of 5-HT and 5-HIAA in both groups are elevated compared with controls. This fact indicates a higher turnover of these substances. The concentration levels are most elevated in parts of the brain of defeated animals exposed to continuous optical contact with the experienced fighter (Stresstiere). These differences are most prominent in the septum and the frontal cortex (s. p. 60).A model for the physiological meaning of these changes is discussed. It was assumed that the activity of the brain is controlled by a system which is functionally analogous to the peripheral autonomous nervous system. 5-HT and norepinephrine should act on antagonistic branches of such a system in central nervous system. Under stress conditions the activity of both branches should be enhanced.

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Herrn Prof. Dr. H. Autrum möchte ich für die Überlassung des Themas, für sein stetes Interesse und anregende Diskussionen danken. Die Untersuchungen wurden durch Sachbeihilfen gefördert, die die Deutsche Forschungsgemeinschaft Herrn Prof. Dr. H. Autrum zur Verfügung stellte.

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Der Firma Sharp & Dohme GmbH danke ich für die kostenlose Überlassung von 15 g Probenecid.     

The effect of clenbuterol on body composition in spontaneously eating tumour-bearing mice

The aim of this study was to investigate the effect of a selective ₂-adrenoceptor agonist, clenbuterol, on body composition in tumour-bearing adult and growing mice. Therefore, adult female C57/BL6 mice (n=20) were inoculated subcutaneously with a 3-methylcholanthrene-induced sarcoma and divided into two identical groups. One group received injections twice a day of clenbuterol corresponding to 1 mg/kg body weight, the other group received sham injections. Growing mice (n=20) were similarly divided after tumour inoculation into one study group with clenbuterol injections and one control group. The growing animals were sacrificed on day 11 after commencement of treatment, the adult mice on day 16.Clenbuterol treatment had no statistically significant effect on accumulated food intake or body composition in the adult mice. However, fooe intake in these animals increased numerically compared to control animals after day 12 of the study. Tumour growth was also unaffected. The growing animals displayed an increased carcass dry weight with borderline significance (p=0.06) and an increased quadriceps muscle fat free dry weight after clenbuterol treatment. Tumour growth was not affected. Food intake measured on a daily basis was significantly increased in the growing clenbuterol treated animals and accumulated food intake was increased with a trend towards statistical significance (p=0.06). The results support the suggestion that treatment with a selective ₂-adrenoceptor agonist does not improve body composition in tumour-bearing adult mice relying on spontaneous food intake while growing animals may benefit from such treatment.     

Effects of fasting and food restriction on sympathetic activity in brown adipose tissue in mice

Summary The activity of the sympathetic nervous system in mice that were either fed ad libitum, food restricted or fasted was estimated by measuring the accumulation of dopamine following the inhibition of dopamine -hydroxylase activity. Mice in each group were injected with the dopamine -hydroxylase inhibitor 1-cyclohexyl-2-mercaptoimidazole and were exposed to either 30°C (warm) or 4°C (cold). Mice were killed 1 h after the injection. Both heart and brown adipose tissue were then quickly removed and homogenized in ice-cold perchloric acid. Dopamine and noradrenaline were determined using high performance liquid chromatography. Regardless of whether mice were warm or cold exposed, both content and concentration of brown adipose tissue and dopamine were predictably higher in 1-cyclohexyl-2-mercaptoimidazole-injected mice than in non-injected animals. In mice fed ad libitum, post-injection content and concentration of dopamine in both brown adipose tissue and heart were higher in cold-exposed mice than in warm-exposed animals. In food-restricted and fasted mice, post-injection concentrations of dopamine in brown adipose tissue were higher in cold-exposed mice than in warm-exposed animals. In food-restricted and fasted mice there was no difference between warm- and cold-exposed animals with respect to post-injection contents and concentrations of dopamine in heart tissue. In fasted mice there was no difference between warm- and cold-exposed animals in post-injection content of dopamine in brown adipose tissue. This study provides further evidence that fasting, in contrast to food restriction, may blunt the tissue sympathetic nervous system response in brown adipose tissue of cold-exposed mice.Abbreviations BAT brown adipose tissue - CHMI 1-cyclohexyl-2-mercaptoimidazole - DA dopamine - DHBA dihydroxybenzylamine - EDTA ethylenediaminetetra-acetic acid - HPLC high performance liquid chromatography - NA noradrenaline - PCA perchloric acid - SNS sympathetic nervous system     

Morphological changes of integumental chloride cells to ambient cadmium during the early development of the teleost, Oreochromis mossambicus

Synopsis Newly-hatched embryos of Oreochromis mossambicus were reared in freshwater and treated with 0 (control), 50 (low level) or 200 (high level) ppb cadmium for 4 days. Changes in the numbers and dimensions of chloride cell apical crypts on the skin of the free embryos were examined daily using scanning electron microscopy. The apical crypts of the chloride cells were rarely observed on the skin of the embryo trunk, and unevenly distributed on the surface of the yolksac. Two days after hatching, the chloride cells of the free embryos exposed to 50 ppb Cd were more active than those of the other two groups. Compared with the control group, the maximum dimensions of the developing apical crypts were stimulated by 50 ppb Cd and inhibited by 200 ppb Cd. The results indicated that the development of chloride cells in tilapia free embryos was provoked by low level Cd exposure and stunted by high level Cd exposure, suggesting the existence of structure/function relationships in which the activation of chloride cells may be related to the ionoregulatory mechanism in adaptation to Cd exposure.     

Metabolism of amino acids during hyposmotic adaptation in the whiteleg shrimp, Litopenaeus vannamei

The penaeid prawn, Litopenaeus vannamei, was employed to investigate intracellular isosmotic regulation in situations where invertebrates encounter hyposmosis. Hemolymph osmolality was first analyzed to confirm osmoregulatory conditions in the experimental animals, followed by analysis of amino acids in muscle and hemolymph using high-performance liquid chromatography. Total muscle amino acid levels decreased when hemolymph osmolality was extremely low, whereas glycine and l-serine levels increased in the hemolymph. These results suggest that tissue amino acids were released into the hemolymph to lower the osmolality of the tissues for purposes of low-salinity adaptation. Next, oxygen consumption and ammonia excretion rates were examined, and the O/N ratio was determined. Oxygen consumption levels and ammonia excretion rates increased, and the O/N ratio decreased when the animals were exposed to low salinity. These results suggest that amino acids were abundantly consumed as an energy source when animals were exposed to low salinity. To confirm the consumption of particular amino acids, the specific activity of l-serine ammonia lyase was also examined. Specific activity was highest when l-serine levels in the hemolymph were highest. Thus, it appears that l-serine levels increased under hyposmotic conditions due to the consumption of l-serine as an energy source. It was concluded that particular amino acids as osmolytes are likely metabolized as energy sources and consumed for purposes of hyposmotic adaptation.     

Photoperiodic Variation in CD45-Positive Cells and Cell Proliferation in the Mediobasal Hypothalamus of the Soay Sheep

The Earth's solar orbit induces annual climatic changes challenging to survival. Many animals have evolved to cope with seasonal variability through compensatory annual changes in their physiology and behavior, which involve innate long-term timing and photoperiodic synchronization to anticipate the environmental seasonal cycles. Here we considered the potential involvement of cyclical histogenesis in seasonal timing mechanisms in the sheep. Adult Soay rams were established in three distinctive seasonal states by controlled photoperiod exposure. A first group, representing the condition in late spring (long-photoperiod [LP] group), was taken indoors in May and exposed to 4 wks of 16?h light/day (LP). A second group was exposed to 20 wks of LP to establish a late-summer/long-day refractory condition (LPR group). A third group of animals was brought indoors in August and exposed to 4 wks of LP followed by 4 wks of 8?h light/day (short photoperiod [SP]) to establish an autumn-like condition (SP group). At the end of these regimes, we injected 5-bromo-2-deoxyuridine (BrdU), and animals were killed 24?h or 4 wks later. When BrdU was administered 24?h before death, more BrdU-immunopositive cells were detected in the hilus of the hippocampus in LP compared with SP animals, indicative of a higher proliferation rate. When BrdU was administered 4 wks before death, more BrdU-positive cells were detected in the hippocampus under LP, compared with SP, indicating increased cell survival. These mitotic cells were occasionally seen to adopt a neuronal phenotype in the hippocampus, but not in the hypothalamus. Approximately 10% of BrdU-positive cells in the basal hypothalamus coexpressed the pan-leukocytic marker CD45, and showed morphological features and regional distribution consistent with ameboid microglia. Increased numbers of these cells were detected in the region of the median eminence and tuberoinfundibular sulcus of animals kept in SP compared with LP or LPR. These data suggest that neuroimmune mechanisms may be involved in photoperiod-dependent seasonal remodeling of the adult brain. (Author correspondence: d.hazlerigg@abdn.ac.uk)     

Alterations in isoforms of glutathione S-transferase in liver and kidney of cadmium exposed rhesus monkeys: Purification and kinetic characterization

Exposure of animals to cadmium (Cd) (25 mg kg-1 body wt day-1) for 10 weeks resulted in preferential accumulation of the metal in liver and kidney. Cd accumulation concomitantly increased zinc (Zn) concentration in both the organs. However, significant decrease in copper level was observed in liver, whereas kidney showed increase in copper (Cu) level. Cd exposure resulted in decreased total GST activity in liver (63%) and kidney (41%) as compared to control group monkeys on normal diet (group I). On isoelectric focusing (IFP) control liver GST segregated into thirteen isoenzymes, while in Cd-treated experimental animals (group II) liver GST resolved into nine isoenzymes. Similarly kidney GST from control animals separated into seven isoenzymes as compared to four isoenzymes from Cd-treated animals. Kinetic analysis showed that Cd exposure did not alter the affinity constant (Km) of GST for GSH and CDNB whereas maximal velocity (Vmax) for these substrates decreased as compared to controls in both the organs, indicating inhibition in GST synthesis by Cd. Cd resulted in a noncompetitive type of inhibition with respect to GSH in vitro. On isoelectric focussing GST of liver and kidney in group II resolved into nine and four isoenzymes as compared to thirteen and seven in group I, showing loss of four basic isoenzymes in case of liver and three isoenzymes in case of kidney. Monkey liver and kidney expressed all the three classes of GST isoenzymes i.e. , µ and , which were serologically identical to human , µ and GSTs. (Mol Cell Biochem 166: 55-63, 1997)