Brain myelination in rats treated with ionizing radiation in utero.

Effects of ionizing radiation on brain myelination and some physical development parameters were studied in laboratory rats (Fisher F-344 inbred strain). Rats were treated with three different doses of radiation (150 rad, 15 rad, and 6.8 rad) delivered on the 20th day of prenatal life. Exposure to 150 rad reduced body, brain, ovary, kidney, heart and spleen weights. Prenatal exposure to 150 rad of radiation reduced the cerebral cortex weight by 22 percent at 30 days of age, and 20 percent at 52 days of age which caused a reduction in cerebral cortex myelin content by 20 and 23 percent at the ages of 30 and 52 days respectively. This dose did not affect the myelin content of the cerebellum or the brain stem, or the myelin concentration (mg myelin/g brain tissue) of the cerebral cortex, cerebellum, and the brain stem. The cerebral cortex weight of the 15 rad treated rats was reduced at the age of 30 days. Exposure to 15 rad, and 6.8 rad did not affect either the myelin content or the myelin concentration of these brain areas.     

Physical and motor development in male and female rat offspring prenatally exposed to gamma radiation

Abstract

The effects of radiation on physical and motor development of male and female rats exposed to ionizing radiation in utero were studied. Rats were exposed to three different doses of radiation (150 rad, 15 rad and 6.8 rad, considered as high, moderate and low doses of radiation respectively) on the 20th day of prenatal life. Exposure to 150 rad contributed to significantly lower body weights of both male and female rat offspring. Upper jaw tooth eruption was delayed in 150 rad treated male offspring, as well as in 15 rad and 150 rad treated female offspring. Cliff‐avoidance response was delayed in 6.8 rad, 15 rad, and 150 rad treated male offspring; and 150 rad treated female offspring. Lower jaw tooth eruption, eye opening, and crawling were not affected by radiation in male or female animals. Results indicate that radiation affected the developmental parameters of both male and female rat offspring, and sex of the offspring played no role in the magnitude of radiation induced damages.     

Mercury induced time-dependent alterations in lipid profiles and lipid peroxidation in different body organs of cat-fish Heteropneustes fossilis

The effects of mercuric chloride (HgCl2) on lipid profiles and lipid peroxidation in different body organs of fresh water cat-fish Heteropneustes fossilis were studied. The daily exposure of HgCl2 0.2 mg/L for 10, 20 and 30 days depleted the total lipids in brain. But the content of phospholipids enhanced significantly at 30 days. Significant diminution in C/P ratio was discernible with 30 days of exposure following mercury toxicosis. Liver exhibited elevated levels of total lipids, phospholipids, cholesterol and C/P ratio. Interestingly kidney showed marked decrease in the concentration of total lipids, cholesterol and C/P ratio at higher exposure. However, the phospholipid values increased during the longer exposure. The content of total lipids and phospholipids was high in muscle but the level of cholesterol and C/P ratio were depleted. Significant increment in lipid peroxidation was discernible in brain, liver and muscle. In kidney the rate of lipid peroxidation was significantly reduced. The results suggest that exposure of HgCl2 enhances the peroxidation of endogenous lipids in brain, liver and muscle. Interestingly the lipid contents are affected differently in different body organs.     

Alterations in the rabbit lymphoid tissue after bendiocarb administration

Various pesticides have immuno-suppressive effects, and thus the organisms become responsive to viral, bacterial and parasitic diseases and neoplasm. The aim of the study was to observe the structure of the small intestine (height of enterocytes and crypts), mucosal lymphoid tissue (Payer's patches, lymphocytes in lamina propria) and a lymph node after administration of bendiocarb (2,2-dimethyl-1,3-benzodioxol-4-yl-methylcarbamate) on days 3, 10, 20, 30 and 60 of the experiment. The height of the observed enterocytes showed an increasing tendency. On days 20, 30 and 60 we also observed an increase in diameter of crypts located in intestinal epithelium. The number of cells in lamina propria mucosae was significantly reduced on days 20 and 30 after administration of bendiocarb. Observations of the lymph node showed that on days 10 and 20 there was a significant increase in relative volume of medulla at the expense of the relative volume of the cortex and a decrease in the number of lymphocytes. However, we recorded an increase in diameter of lymphocytes. The intracellular parasite Toxoplasma gondii (T. gondii) belongs to the most common pathogenic parasites in the world and it can cause serious health complications in pregnant and immunodeficient individuals. DNA isolation, standard polymerase chain reaction (PCR) and visualization in a 2.5 % agarose gel, the presence of DNA T. gondii was detected in no examined rabbit brain samples. Using real time PCR T. gondii DNA was detected and quantified in the three rabbit brain samples (10 %).     

Exposure to sublethal concentrations of Zn(II) and Cu(II) changes biochemical parameters in Leporinus obtusidens

The aim of the present study was to assess the effect of the exposure of Leporinus obtusidens (Piava) to zinc and copper on catalase activity in the liver, delta-aminolevulinate dehidratase (delta-ALA-D) activity in liver, muscle, brain and kidney, and thiobarbituric reactive species (TBARS) in brain, muscle and liver. In addition, hematological parameters were measured in blood. The fish were exposed to 10% and 20% of the derived LC(50) values, 2.3 and 4.6 mg Zn l(-1) and 0.02 and 0.04 mg Cu l(-1), and sampled on days 30 and 45. Exposure to Zn(II) and Cu(II) decreased hematological parameters and also delta-ALA-D activity mainly in liver and kidney at all concentrations tested. Liver catalase activity increased after zinc or copper exposure at all concentrations and exposure times tested. Thiobarbituric reactive substances (TBARS) increased in the brain and liver of the fish exposed to zinc(II) for 45 days at both metal concentrations. In muscle, zinc(II) increased TBARS production at both exposure times and concentrations tested. Copper(II) exposure reduced the TBARS levels in liver at both concentrations and times tested. In brain, there was a decrease in TBARS levels only after 45 days of exposure. In muscle, this decrease was observed after 30 days of exposure at both concentrations. Although zinc and copper are required as microelements in the cells, our results showed that the sublethal concentrations of these metals can change biochemical parameters which may alter normal cellular function. These results pointed out the differential sensitivity of fish tissues to essential, but also toxic and environmentally relevant metals. The alterations of distinct biochemical parameters in fish tissues certainly contribute to the toxicity of Zn and Cu, and are of importance for an area that has been growing and has still been poorly explored in the literature.     

Fluoride induced changes in protein metabolism in the tissues of freshwater crab Barytelphusa guerini

Exposure of the freshwater field crab Barytelphusa guerini to a sublethal concentration of NaF (30 mg litre(-1)) caused significant alterations in protein metabolism. After an exposure time of 15 days, the crab was found to have a marked depletion of total protein and free amino acid content. A significant elevation in transaminases, Aspartate aminotransferase (AAT) and Alanine aminotransferase (ALAT), and Glutamate dehydrogenase (GDH) activities was reflected in the free amino acid levels of the tissues.     

Effect of the trichothecene deoxynivalenol on brain biogenic monoamines concentrations in rats and chickens

Male Sprague-Dawley rats (180 g) and 28-day-old Single Comb White Leghorn Cockerels (300 g) were orally dosed with deoxynivalenol (DON) at 2.5 mg kg-1 body weight. In the first experiment, whole brains were collected at 2, 6, 12, 24 and 48 hours after the toxin treatment and analyzed for brain biogenic monoamines by high-performance liquid chromatography with electrochemical detection. Although several interesting trends were observed, DON did not influence whole brain concentrations of monoamine neurotransmitters or their metabolites in either species, at any time. In a second experiment, brains were collected 24 hours postdosing, dissected into 5 brain regions (pons and medulla oblongata, cerebellum, hypothalamus, hippocampus and cerebral cortex), and analyzed. DON treatment resulted in significantly elevated concentrations of serotonin (HT) and 5-hydroxyindole-3-acetic acid (HIAA) in all brain regions of the rat. However, this was not seen in poultry, where DON treatment resulted in a decrease in norepinephrine (NE) in the hypothalamus and hippocampus, and a decrease in dopamine (DA) in the pons and medulla oblongata region. These results suggest that DON influences brain biogenic amine metabolism, and that there may be intraspecies differences in the central effects of this mycotoxin.     

Effects of dioxins and polychlorinated biphenyls (PCBs) on thyroid function in infants born in Japan – the second report from research on environmental health

Dioxins (PCDD + PCDF) and polychlorinated biphenyl (PCB) are potentially hazardous compounds and have structural similarity to thyroid hormones. Our research group on “Dioxins and PCB in Human Milk” was organized in 1997 and has been active for the past three years. We collected breast milk from 80 mothers living in Tokyo, Saitama, Ishikawa and Osaka Prefecture in Japan at 5, 30, 150, 300 postpartum days and measured PCDDs, PCDFs and PCBs during 1998–1999. We added another 20 prefectures and cities during 1999–2000. Breast milk was obtained from 415 mothers at 30 postpartum days (breast-feeding group). Blood was taken from infants who were breast-fed at the age of 1 year for evaluation of thyroid and immune functions. Blood was also taken from 53 infants who were bottle-fed (bottle-feeding group) as a control. Serum T4, T3, FT4 and TSH levels in the breast-feeding (n=337) and bottle-feeding (n=53) groups were within normal ranges and were not significantly different between the two groups. Although there were a large geographic differences of dioxins and co-PCB content in breast milk, there were no differences in serum T4, T3, FT4, TSH levels and no significant correlation between the mean serum levels of TSH and TEQ in breast milk. There was a significant correlation between serum TSH at 1 year of age and that from blood on dried filter paper TSH at 5 days of age. We concluded that dioxin intake from breast milk in Japanese infants did not impair thyroid function. However, long-term effects remain to be evaluated.     

Polychlorinated dibenzo-p-dioxins and dibenzofurans in commercial diphenyl ether herbicides,and in freshwater fish collected from the application area

This paper describes the analyses of polychlorinated dibenzodioxins, dibenzofurans, and related compounds in diphenyl ether herbicides, CNP, NIP, and X-52. The levels of tetra-, penta-CDDs, and tetra-CDFs were about 150, 30, and 15 ppm, respectively in CNP formulations. NIP and X-52 contained lower chlorine alalogs. Isomer distribution of PCDDs in CNP agreed with an estimate from the reaction of chlorophenols. Freshwater fish samples heavily contaminated with CNP in the application season also contained 0.2 ppb of 1,3,6,8-tetra-CDD.     

Alteration in some biochemical and enzymological parameters in the snake head fish , exposed chronically to quinalphos

The effect of chronic quinalphos exposure (0.025 mg/1) for 15 and 30 days on the levels of glucose, lactic acid and haemoglobin in the blood; glycogen and lactic acid contents of the liver and muscles; and the activities of hexokinase, lactate dehydrogenase, pyruvate dehydrogenase and succinate dehydrogenase in liver, kidney, intestine, brain, gills and muscles was examined. Blood glucose, lactic acid and haemoglobin levels decreased in quinalphos exposed fish. Glycogen content of liver and muscles increased but lactic acid decreased. Hexokinase was inhibited in intestine and muscles after 30 days of exposure but increase in enzyme activity was noted in gills. Lactate dehydrogenase activity was inhibited in all the six tissues. Pyruvate dehydrogenase activity of liver, kidney, gills and muscles was inhibited. However, in brain the enzyme activity was elevated. Succinate dehydrogenase activity was elevated in intestine and inhibited in other tissues.     

Soil dehydrogenase, phosphomonoesterase and arginine deaminase activities in an insecticide treated groundnut (Arachis hypogaea L.) field

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2 pyridyl phosphorothioate) 20 EC and Quinalphos (O,O-diethyl O-quinoxalin-2-yl phosphorothioate) 25 EC, were applied in groundnut (Arachis hypogaea L.) field as seed treatment at 25 ml/kg and soil treatment at 4 l/ha in 1998 and 1999. The residues of these insecticides were monitored during the entire crop season and their effect on the soil enzymes dehydrogenase, phosphomonoesterase and arginine deaminase were studied. Ninety nine percent of chlorpyrifos residues were dissipated within 60 days from seed treated soil and 98% dissipation was observed in soil treated field for the same days. Its half lives in seed treated soil were 8 days and 7 days and in soil treated field were 9.2 days in and 7.5 days in 1998 and 1999 respectively. Dissipation of quinalphos in comparison to chlorpyrifos was slow both in seed treated and soil treated field. Eighty seven percentage to 92% dissipation of quinalphos residues were observed from seed treated soil and 98% residues were dissipated from soil treated field within 75 days. Its half lives in seed treated soil were 20 days and 18 days and in soil treated field, its half lives were 13 days and 17 days 1998 and 1999 respectively. Inhibition in dehydrogenase activity followed by recovery was observed both in seed and soil treatments with chlorpyrifos. An inhibition of 17.2% was estimated after 60 days of seed treatment in comparison to control. Dehydrogenase activity was significantly reduced to 63% after 15 days of quinalphos seed treatment in comparison to control in 1998. Similar trends were observed in 1999. A significant inhibition in dehydrogenase activity was observed after soil treatment both in 1998 and 1999. Phosphomonoesterase activities were significantly inhibited upto 25.2% as compared to the control, on the 15th day of chlorpyrifos seed treatment in 1998 and similarly, after one day of treatment in 1999. Quinalphos inhibited the phosphomonoesterase activity till the end of the experimental period in the soil treated fields, whereas recovered within 30-60 days of treatment in the seed treated fields. Arginine deaminase activity was significantly stimulated within one day after chlorpyrifos seed and soil treatments in both years. The activity was almost threefold higher on the 30th and the 15th day of soil treatment in 1998 and 1999, respectively. A temporary inhibition of arginine deaminase activity was observed after quinalphos treatment. It was observed that in most of cases insecticides have temporary inhibitory effect on soil enzymes. However, inhibition was smaller in seed treated soil than in direct soil treatment.     

Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats

Two experiments were conducted in male SD rats (225-250 g) to determine changes in the activities of endogenous antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and concentrations of glutathione (GSH) in tissues after exposure to low doses of endosulfan and chlorpyrifos using a whole body exposure technique. In both experiments, 6 rats/group were exposed 3 hr/day, 5 days/week for 30 days to: 0 (control), 5, 10, 20, 40 and 60% of LD50 of either pesticide in 50% ethanol; actual concentrations were: endosulfan = 0, 0.5, 1.0, 2.0, 4.0, 6.0 mg/250 g body weight; chlorpyrifos = 0, 1.9, 3.8, 7.6, 15.2, and 22.8 mg/250 g body weight. Endosulfan decreased erythrocyte SOD by 21% in all groups and chlorpyrifos increased SOD by 18% in groups 40 and 60. Liver SOD was 12%-20% lower after endosulfan exposure; lung SOD was altered: endosulfan decreased activity by 21% and 51% and chlorpyrifos by 58 and 75% in the 40 and 60 groups, respectively (P < or = 0.05). Both pesticides increased plasma GPX activity at lower levels and reduced it by 26% and 19% in groups 40 and 60, respectively (P < or = 0.05). Liver GPX increased in the 60 group and lung GPX declined between 20% and 38% after endosulfan exposure. GSH in the liver and lung: endosulfan reduced GSH by about 30% at lower levels and increased by 41% or 70% at higher levels; chlorpyrifos decreased GSH by 28-40% in 20 and 60 groups, respectively (P < or = 0.05). Exposure to low, increasing levels of endosulfan and chlorpyrifos can differentially modify endogenous antioxidants SOD, GPX and GSH, which may lead to the development of oxidative stress in some tissues.     

Alteration in some biochemical and enzymological parameters in the snake head fish Channapunctatus, exposed chronically to quinalphos

The effect of chronic quinalphos exposure (0.025 mg/1) for 15 and 30 days on the levels of glucose, lactic acid and haemoglobin in the blood; glycogen and lactic acid contents of the liver and muscles; and the activities of hexokinase, lactate dehydrogenase, pyruvate dehydrogenase and succinate dehydrogenase in liver, kidney, intestine, brain, gills and muscles was examined. Blood glucose, lactic acid and haemoglobin levels decreased in quinalphos exposed fish. Glycogen content of liver and muscles increased but lactic acid decreased. Hexokinase was inhibited in intestine and muscles after 30 days of exposure but increase in enzyme activity was noted in gills. Lactate dehydrogenase activity was inhibited in all the six tissues. Pyruvate dehydrogenase activity of liver, kidney, gills and muscles was inhibited. However, in brain the enzyme activity was elevated. Succinate dehydrogenase activity was elevated in intestine and inhibited in other tissues.     

Rare earth elements in naturally grown fern Dicranopteris linearis in relation to their variation in soils in south-Jiangxi region (southern China)

Ferns (Dicranopteris linearis) and soils, sampled from four rare earth mining areas and one non-mining area locating in South-Jiangxi region, were chosen for analysis of 15 rare earth elements (REEs) by inductively coupled plasma-mass spectrometry (ICP-MS). The fern samples were divided into 4 parts: root, stem, petiole, and lamina. The soils of the same sites were also sampled, and 4 soil layers were observed at the profile: A (0-20 cm depth), B (20-150 cm depth), C (150-400 cm depth), and D (rock). The distribution patterns and transportation characteristics of REEs of different soil layers and of different parts of D. linearis were studied. The results showed that in the soil layers, the sigma REE (the sum of the concentrations of 15 REEs) of A layer, as well as B layer were lower than that of C layer, but Ce was relatively concentrated in A layer. The sigma REE of different parts of D. linearis were lamina, root > stem > petiole. The REE distribution patterns of D. linearis root were similar to the total REE distribution patterns of the A layer of their host soils, especially to the soluble REE distribution patterns of the A layer of their host soils. The REEs were fractionated in their transportation in D. linearis. The abundances of heavy rare earth elements (HREEs, Gd to Lu and Y) in the stem, petiole, lamina are lower than those in the root.     

Cobalt-induced oxidative stress in brain, liver and kidney of goldfish Carassius auratus

Cobalt is an essential element, but at high concentrations it is toxic. In addition to its well-known function as an integral part of cobalamin (vitamin B₁₂), cobalt has recently been shown to be a mimetic of hypoxia and a stimulator of the production of reactive oxygen species. The present study investigated the responses of goldfish, Carassius auratus, to 96 h exposure to 50, 100 or 150 mg L⁻¹ Co²⁺ in aquarium water (administered as CoCl₂). The concentrations of cobalt in aquaria did not change during fish exposure. Exposure to cobalt resulted in increased levels of lipid peroxides in brain (a 111% increase after exposure to 150 mg L⁻¹ Co²⁺) and liver (30-66% increases after exposure to 50-150 mg L⁻¹ Co²⁺), whereas the content of protein carbonyls rose only in kidney (by 112%) after exposure to 150 mg L⁻¹ cobalt. Low molecular mass thiols were depleted by 24-41% in brain in response to cobalt treatment. The activities of primary antioxidant enzymes, superoxide dismutase (SOD) and catalase, were substantially suppressed in brain and liver as a result of Co²⁺ exposure, whereas in kidney catalase activity was unchanged and SOD activity increased. The activities of glutathione-related enzymes, glutathione peroxidase and glutathione-S-transferase, did not change as a result of cobalt exposure, but glutathione reductase activity increased by ∼40% and ∼70% in brain and kidney, respectively. Taken together, these data show that exposure of fish to Co²⁺ ions results in the development of oxidative stress and the activation of defense systems in different goldfish tissues.     

Toxicity of copper on rice growth and accumulation of copper in rice grain in copper contaminated soil

Pot soil experiments showed that copper (Cu) is highly toxic to rice. Rice grain yields decreased exponentially and significantly with the increase of soil Cu levels. Rice grain yield was reduced about 10% by soil Cu level of 100 mg kg(-1), about 50% by soil Cu level of 300-500 mg kg(-1) and about 90% by soil Cu concentration of 1,000 mg kg(-1). Root was more sensitive to soil Cu toxicity than other parts of rice plant at relatively lower soil Cu levels (less than 300-500 mg kg(-1)), but the growth of whole rice plant was severely inhibited at high soil Cu levels (300-500 mg kg(-1) or above). Cu concentrations in rice grain increased with soil Cu levels below 150-200 mg kg(-1), but decreased with soil Cu levels above 150-200 mg kg(-1), with peak Cu concentration at soil Cu level of 150-20 mg kg(-1). Cu was not distributed evenly in different parts of rice grain. Cu concentration in cortex (embryo) was more than 2-fold that in chaff and polished rice. More than 60% of the Cu in grain was accumulated in polished rice, about 24% in cortex (embryo), and about 12% in chaff. So, about 1/3 of the Cu in rice grain was eliminated after grain processing (chaff, cortex and embryo was removed).     

Effect of manganese on the concentration of amino acids in different regions of the rat brain

The present study was designed to determine if chronic exposure of weanlings and adult rats to Mn produces significant alterations in amino acid concentrations in different regions of the rat brain. Weanling (30 day old) and adult (90 day old) male rats were exposed to 10 and 20 mg Mn/kg body weight per day, by gavage, for 30 days. Forty-eight hours after the last dose, animals were sacrificed by decapitation and brains were dissected into different regions to determine the concentration of amino acids by HPLC/EC. A dose dependent decrease in body weight gain was found in the adult, but not in the weanling rats. Significant increases occurred in concentrations of aspartate, glutamate, glutamine, taurine and gamma-aminobutyric acid (GABA) in the cerebellum of the adult rats dosed with 20 mg/kg per day, Mn. A significant decrease in the concentration of glutamine was observed in caudate nucleus and hippocampus of weanling rats dosed with 10 mg/kg, Mn. These data suggest that chronic Mn exposure can produce a decrease in body weight gain in adult rats and alterations in amino acids in different regions of weanling and adult rat brains.     

Influence of pH and aluminum on developing brook trout in a low calcium water

Eyed embryos of brook trout (Salvelinus fontinalis) were exposed to nominal pHs of 4.5, 5.5 and 7.5 with and without aluminum (300 microg liter(-1)) in extremely soft water (hardness <9mg liter (-1)) at 12 degrees C. Embryo mortality exceeded 80% at pH 4.5, averaged 15 to 18% in the pH 5.5 treatments and was less than 2% in the pH 7.5 treatments. Aluminum significantly reduced embryo mortality (85.3% vs 99.5%) at pH 4.5 but did not affect mortality at pH 5.5 or pH 7.5. Percent hatch and poor hatch were pH dependent and were not significantly influenced by aluminum. Brook trout larvae cumulative mortalities were 100% within 30 days at pH 4.5, with or without aluminum; 69% after 60 days at pH 5.5; 100% in 15 days at pH 5.5 with aluminum and 20% after 60 days at pH 7.5 with or without aluminum. Fish that survived the pH 5.5 treatment showed decreased growth and behavioral impairments compared to the controls (pH 7.5 without aluminum).     

28-homobrassinolide protects chickpea (Cicer arietinum) from cadmium toxicity by stimulating antioxidants

In the present experiment the seeds of Cicer arietinum (L.) cv. Uday were inoculated with specific Rhizobium grown in sandy loam soil and were allowed to grow for 15 days. At this stage, the seedlings were supplied with 0, 50, 100 or 150 microM of cadmium in the form of cadmium chloride and sprayed with 0.01 microM of 28-homobrassinolide (HBL) at 30-day stage. The data indicated that plant fresh and dry mass, number of nodules, their fresh and dry mass, leghemoglobin content, nitrogen and carbohydrate content in the nodules, leaf chlorophyll content, nitrate reductase and carbonic anhydrase activities decreased proportionately with the increasing concentrations of cadmium but the content of proline and the activities of catalase, peroxidase and superoxide dismutase increased. The ill effect, generated by cadmium, was overcome if the stressed plants were sprayed with HBL.     

Effect of manganese on the concentration of amino acids in different regions of the rat brain

Abstract

The present study was designed to determine if chronic exposure of weanlings and adult rats to Mn produces significant alterations in amino acid concentrations in different regions of the rat brain. Weanling (30 day old) and adult (90 day old) male rats were exposed to 10 and 20 mg Mn/kg body weight per day, by gavage, for 30 days. Forty‐ eight hours after the last dose, animals were sacrificed by decapitation and brains were dissected into different regions to determine the concentration of amino acids by HPLC/EC. A dose dependent decrease in body weight gain was found in the adult, but not in the weanling rats. Significant increases occurred in concentrations of aspartate, glutamate, glutamine, taurine and y‐aminobutyric acid (GABA) in the cerebellum of the adult rats dosed with 20 mg/kg per day, Mn. A significant decrease in the concentration of glutamine was observed in caudate nucleus and hippocampus of weanling rats dosed with 10 mg/kg, Mn. These data suggest that chronic Mn exposure can produce a decrease in body weight gain in adult rats and alterations in amino acids in different regions of weanling and adult rat brains.