Effects of the spaceflight on organ-development in the neonatal rats: results in the Neurolab (STS-90)]

In the Neurolab mission, we found that spaceflight affects the development of the aortic baroreflex system and the body weight of the flight rats was significantly lighter [correction of lightess] than that of the control group. The aim of this study is to examine the structural and functional development in various tissues and organs. One hundred and eighteen nine-day old rats and seven fifteen-day old rats, which were launched at these ages and nursed by their dams in the space shuttle Columbia for 16 days, were served for this study. Two hundred and twenty one neonates were used as the ground controls (VIV: vivarium and AGC: asynchronous ground controls). On the landing day after they returned to the earth, the rats were perfused with a fixative under deep urethane anesthesia, and the organs were weighed and the ratio of the organ weight to the body weight was calculated. Six animals of the nine-day old group were reared on the ground for 30 more days after landing and also examined in the same protocol as the landing-day-examination. The organs obtained to examine were heart, lung, spleen, thymus, adrenal glands, kidney, liver, small intestine, large intestine, mesentery, pancreas, testis and ovary. Paraffin sections were made from some organ tissues and prepared for HE staining and immunohistochemistry. We compared these organs in the flight rat with those in the ground controls. All organs except the lung of nine-day old group were significantly smaller. In the ratio of organ weight to body weight, the lung and heart were significantly larger. The weight and ratio of the liver showed no significant difference. The thymus, spleen, mesentery and pancreas were smaller in the weight and the ratio. There were no differences in the body weight among 30-day reared groups, but the lung in the flight group is significantly heavier than the control groups and thymus also tends to be relatively heavy. In flight rats of the fifteen-day group, the kidney was heavy and the ovary was light as compared to the controls. The adipose tissue was macroscopically little found around the thoracic and abdominal organs in all rats of the flight group. These results suggest that the organs related to oxygen supply like as the lung and heart have priority in development over the mesentery and immune system organs even during spaceflight. Lightness of the mesentery in space rats is due to small contents of adipose tissues, and may reflect amounts of the food taken by the flight dams. Lightness of the organs like as the thymus, spleen and pancreas suggests that spaceflight may affect the immune system and also affect continuously the lung and thymus development even after landing.     

Morphological characteristics of the kidney and lung in the neonatal rats observed after 16 days spaceflight.

The aim of this study is to examine the structural development in kidney and lung macroscopically which relate with cardiovascular system in rats raised in space. Twenty three nine-day old rats and six fifteen-day old rats, which were launched at these ages and nursed by their dams in the Space Shuttle Colombia for 16 days (STS-90; Neurolab). Seventeen animals of the nine-day old rats were defined as the nine-day group, and the rest was defined as the re-adaptation group, which were reared on the ground for 30 more days after landing. The organs were weighed and the ratio of the organ weight to the body weight (body weight ratio) was calculated. Both of lung and kidney in flight rats were significantly heavier than ground controls in the body weight ratio. We found that the kidney in the nine-day and the fifteen-day group tended to extend of dorsal-ventral length in macroscopic observations. However, this difference was not observed in the re-adaptation group. These results suggest that space environment may affect in kidney development. On the other hand, the lung had no differences in macroscopic structure among flight and control groups.     

Dynamics of organ blood flow in rats in postnatal ontogenesis

In the 4-, 13-, 30-, and 65–90-day old anestetized Wistar rats (8–15 rats in each group) the voluminous blood flow rate was measured in liver, kidney, small intestine wall, and in the predominantly white gracilis muscle of femur. A LAKK-01 laser-Doppler flowmeter and its cutaneous (for kidney) and needle (for other organs) probes were used; they provided estimation of blood flow to the tissue depth of about 1 mm. It has been found that the blood flow rates (per organ mass unit) fall in liver, intestine, and even more in muscle, whereas increase in kidney, particularly for the first month of life. Calculations show that in adult rats as compared with the 4-day old pups (with their mass exceeding 22 times) the blood flow to muscle, liver, intestine, and kidney should rise (with taking into account the increase of mass of the organs) 14, 17, 18, and 43 times, respectively.     

Repair processes of hemopoiesis after applying cyclophosphamide. II. Quantitative changes in basic proteins and DNA in the bone marrow, spleen, and thymus

The effect of cyclophosphamide on the organ weight, DNA and basic protein content in nuclei of bone marrow, thymus and spleen of rats was studied. Animals were examined 6, 12 and 24 hours and 3, 5, 10, 20 and 30 days after intraperitoneal application of cyclophosphamide in the doses of 100 and 200 mg X kg-1 of body weight. From the 5th day after application a marked decrease in weight and cellularity of all organs was observed. Amounts of DNA and basic proteins in nuclei of bone marrow and spleen were higher as compared with controls and their increase occurred prior to recovery of cellularity and organ weight, that was initiated within 5 and 10 days after cyclophosphamide treatment. Changes in the thymus persisted for a longer period of time and recovery was incomplete even at the day 30 after cyclophosphamide application. In accordance with these data no increase in DNA and basic protein amounts in the thymus nuclei was observed.     

Leptin-programmed rats respond to cold exposure changing hypothalamic leptin receptor and thyroid function differently from cold-exposed controls

We showed that neonatal leptin treatment programmes for hyperleptinemia and central leptin resistance both at 30days-old and adulthood, while programmes for lower serum T3 at 30days-old, but higher thyroid hormones (TH) at adulthood. As in these animals, acute cold at 30days-old normalized leptinemia and restored the expression of hypothalamic leptin receptor (OBR), here we evaluate the effect of cold exposure on the thyroid function and OBR in adult rats programmed by neonatal hyperleptinemia. Pups were divided into 2 groups: Lep-injected with leptin (8μg/100g/BW, sc) for the first 10days of lactation, and C-injected with saline. At 150days, both groups were subdivided into: LepC and CC, which were exposed to 8°C for 12h. Serum leptin, TH, TSH, liver type I and brown adipose tissue (BAT) type II deiodinases (D1 and D2) activities, liver mitochondrial alpha-glycerol-3-phosphate dehydrogenase (mGPD) activity and adrenal catecholamine content were measured. Hypothalamic and thyroid OBR protein contents were evaluated. Differences were significant when p<0.05. Lep group had hyperleptinemia (+19%), higher T4 (+20%) and T3 (+30%) with lower TSH (-55%), higher liver D1 (1.4 fold-increase), lower BAT D2 (-44%) and liver mGPD activities (-55%), higher adrenal catecholamines (+44%), lower hypothalamic OBR (-51%) and normal thyroid OBR. Cold exposure normalized leptinemia, D1, mGPD, catecholamine and hypothalamic OBR. However, cold exposure further increased TH and decreased D2. Thus, cold exposure normalizes most of the changes programmed by neonatal hyperleptinemia, at the expense of worsening the hyperthyroidism and BAT thermogenesis.     

Acute and chronic hypoxia in rats. I. Effect on organismic respiration, mitochondrial protein mass in liver and succinic dehydrogenase activity in liver, kidney and heart

The objective of this investigation was to characterize organismic, organ and mitochondrial alterations in rats over the course of 27 days at 0.4 atm. In the adjustment phase (day 1 through 5) a significant decrease in systemic oxygen uptake and body weight (23% of pre-altitude values) occurred. In the acclimating state (day 7 to 27) body weight was regained but oxygen consumption remained depressed. Hematocrit increased hyperbolically from 45% in 0-day rats to 79% in 27-day rats. Liver, kidney and heart weights and total organ protein paralleled the changes observed in body weight. Total organ succinic dehydrogenase activity showed a wave-like oscillation for liver and kidney; activity was decreased in both organs by day 5, showed a transient but significant increase on days 16 through 18 and a return to diminished activity on day 27. Succinic dehydrogenase activity for heart became depressed in the adjustment phase but showed a stable level comparable to pre-altitude values in the accliminating phase, days 7 through 27. Liver mitochondrial protein mass was unchanged from pre-altitude values on days 5 and 27 even though succinic dehydrogenase activity was significantly depressed. Therefore, the changes in succinic dehydrogenase activity are not representative of altered mitochondrial mass but suggest that mitochondrial function was altered.     

The development of the lymphoid organs of flounder, Paralichthys olivaceus, from hatching to 13 months

The growth of the lymphoid organs, such as head kidney, spleen and thymus were studied in flounder, Paralichthys olivaceus Temminck & Schlegel, from hatching to 13 months of age. Except for the thymus, all organs grew as the fish grew. By 2 months of age the lymphoid organs attained their maximum relative weight. The organ weight showed a closer correlation to body weight than they did to age. The total number of leucocytes in the lymphoid organs increased with age, but the number per milligram of lymphoid organ remained constant. A micro and ultrastructural study of the lymphoid organs showed that the full development of the lymphoid organs was not achieved until the juvenile stage. The spleen and head kidney had mixed populations of "red" and "white" cells. The head kidney was more lymphoid than the spleen. The thymus involuted quickly during the first 6 months. The blood components had no obvious relationship with age or season during the period studied.     

Growth enhancement induced by prolonged L-dopa administration in rats

To follow growth of rats, in which growth hormone secretion has been chronically stimulated, L-Dopa (5 mg/kg) was injected subcutaneously twice daily for 70 days to growing rats. A control group, matched for sex and sibship, pair fed with the treatment group was given saline injections. At 10-day intervals, the rats were weighed and measured. At 90 days of age the rats were ether anesthesized, bled for growth hormone determination, and internal viscera weighed. Weight gain and length in the L-Dopa-treated group was found to be significantly greater. A mean weight gain of 6% and 12% in the male and female rats, respectively, and a mean length gain of 5% in both male and female rats was observed at 90 days of age. The thymus, thyroid, adrenals, uterus, and gonads all tended to be heavier in the L-Dopa-treated group. Significantly heavier kidneys were found in the L-Dopa group. Serum growth hormone was found to be 8.44 +/- 1.4(SE) ng/ml in the L-Dopa group and 4.6 +/- 0.9 ng/ml in the control group. It is concluded that the continuous administration of L-Dopa produces an increase of circulating serum growth hormone levels, and this in turn enhances growth.     

洞庭湖区社鼠脏器重量的比较

对洞庭湖区社鼠(Niviventer confucianus)野外自然种群脏器的重量指标进行了测定,并比较了其在年龄组、性别、季节及生境间的差异。结果表明,社鼠内脏(心、肺、肝、脾、肾脏)随着年龄组的增加,重量有明显的增加,其重量与体重有极其显著的相关性。两性间的脏器重量指标没有显著性差异。脏器季节变化的共同特征是夏季脏器重量较低,四季间比较,仅有心脏重量有显著的季节变化。生境间心脏和肾脏重量的变化相对较大,达显著水平。参与繁殖与未参与繁殖的雌鼠相比,心、肺、肝、肾脏的各项指标均较高,脾脏则相反,但均未有显著性差异。总的来看,洞庭湖社鼠种群的脏器指标相对稳定,尽管重量指标随着年龄组而增加,受性别、季节、生境及繁殖行为的影响相对较小。     

Metabolism of zinc and copper in the neonate: Zinc thionein in developing rat brain,heart, lung,spleen, and thymus

In a continuing study of the importance of metallothionein (MT) in the growth and development of neonates, zinc and copper metabolism in rat brain, heart, lung, spleen, and thymus has been analyzed in 5, 10, 15, 20 and 25 day old rats. Total, cytosol, and MT zinc and copper concentrations and organ contents were determined. Zinc, but very little, if any copper was associated with MT in these organs. Concentrations ranged from 0.03 to 3.3 μg Zn in MT/g; organ contents ranged from 0.003 to 2.2 μg Zn in MT/organ. Brain exhibited the highest concentrations and contents of zinc in MT, approaching the levels found in kidneys. Rank order of organ contents of zinc in MT was brain > lung > heart, spleen, thymus, during this neonatal growth period. When organ growth was rapid, a large percentage (20–95%) of the cytosolic zinc present in these organs was associated with MT, as has been previously observed with liver, kidneys, and testes. None of these organs undergoes the dramatic changes in zinc and copper metabolism previously observed in neonatal rat liver and gastrointestinal tract, and in maturing testes. They are more comparable to kidneys in their concentrations of zinc in MT. Like testes, little copper is found in these organs.     

Stimulation of ornithine decarboxylase activity in rat tissues by growth hormone and by serum growth factor from rats infested with spargana of Spirometra mansonoides

Ornithine decarboxylase activity was studied in heart, kidney, liver, thymus, lung, spleen, skeletal muscle and fat of hypophysectomized rats after growth hormone treatment. A marked increase in enzyme activity was observed in kidney and liver, and a significant increase in heart and thymus at 4 h after injection of growth hormone. The kidney was the most responsive organ with an increase in activity of about 100 fold. The enzyme activity in kidney responded to a dose of 10 μg of growth hormone. Daily injection for 12 days raised activity only in the heart. Infestation for 6–13 days with spargana of Spirometra mansonoides, which also causes growth of hypophysectomized rats, increased enzyme activity in the heart and thymus. Intravenous injection of serum of hypophysectomized rats infested with spargana of Spirometra mansonoides caused a significant increase in the enzyme activity in liver and kidney after 4 h. Growth hormone and the serum growth factor of sparganosis seem to share the characteristic of causing an early increase in ornithine decarboxylase activity in rat tissues. The marked response in kidney and liver raises the possibility that these organs are the primary targets of both substances.     

Influence of castration on development of the thymus in neonatal male rats.

The influence of castration on the development of the thymus in neonatal rats was studied to elucidate when after birth the thymus comes under inhibitory regulation by the testis in rats. The relative and absolute weights of the thymus were measured five days after castration these cases. No excessive changes in the weights of the thymus with castration were observed by 31 days after birth. Significant changes in the thymus appeared in the relative weight at 36-day-castration. The absolute weight of the thymus was also significantly increased after 41-day-castration. These findings suggest that in rats the inhibitory regulation of the thymus by testis development does not appear before at least 31 days of age.     

Differential recovery of intestine, bone marrow, and thymus of rats with solid tumors following 5-fluorouracil administration.

Time relationships for recovery of several host organs from toxic effects of 5-fluorouracil were determined in ACI rats bearing Morris hepatoma 3924A. A single injection of 150 mg/kg body weight 5-fluorouracil (the LD10) resulted in loss of 90% of the tibial bone marrow, 60% of the intestinal mucosa, and 90% of the thymus as measured by total DNA content of the organs. Organ DNA contents following 150 mg/kg of the drug were minimal on day 3 for intestine and on day 5 for marrow and thymus. A return to pretreatment or higher levels of DNA was observed by day 4 for intestine, day 11 for tibial marrow, and day 19 for thymus. Incorporation of 3H-deoxyuridine into host organ DNA after 150 mg/kg 5-fluorouracil was inhibited 36 hrs for intestine, 3 days for thymus, and 5 days for tibial bone marrow. Inhibition of 3H-deoxyuridine incorporation into DNA was similar for 50, 100, and 150 mg/kg doses both in tumor and in host organs, but recovery of 3H-deoxyuridine incorporation and DNA content of host organs began later with the higher doses of 5-fluorouracil. Maximal incorporation of 3H-deoxyuridine into DNA was observed on day 4 for intestine, day 8 for marrow, and day 9 for thymus after treatment with 150 mg/kg 5-fluorouracil. Animal lethality following the second of two 150 mg/kg injections of 5-fluorouracil was related to the extent of recovery of intestinal mucosa and bone marrow at the time of the second injection. Survival decreased to 0% for normal rats when the interval between injections was 3-4 days, improved at 5 days and was 100% when the interval was 10-11 days.     

Thymus, peripheral lymphoid tissues and immunological responsiveness of the pituitary dward mouse (author's transl)]

Snell's pituitary dwarf mice (dw) were used for studies on the relationship between hypophysis and lymphoid organs. The age-dependent changes of thymus or spleen weights of dwarf mice were compared with those of normal littermates. The suppression of growth of the thymus or spleen in dwarf mice was recognized at 5th day of age. Although involution of the thymus varied among animals, a strong positive correlation was demonstrated between relative thymus weight and body weight in 30 approximately 40 days old dwarf mice. Lymphoid organs of dwarf mice were reconstituted by injection of growth hormone and or thyroxin. Relative thymus weight significantly increased in dwarf mice when the treatment with growth hormone started at 7 days of age, but the same treatment at 3 months of age did not show any effect on the increment of relative thymus weight. On the other hand, the antibody-forming capacitiy against sheep erythrocytes of dwarf mice was significantly increased even when the treatment with growth hormone was started at 3 months of age. A marked increase in the number of lymphoid cells in dwarf mice was observed by treatment with thyroxin, even if treatment was started either at 7 days or 3 months of age. Similar changes were also obtained in the antibody-forming capacity.     

Effects of strenuous maternal exercise on fetal organ weights and skeletal muscle development in rats

The purpose of the present study was to observe the effects of strenuous maternal aerobic exercise throughout gestation on fetal outcome in the rat. The strenuous exercise intensity consisted of a treadmill speed of 30 m.min-1 on a 10 degrees incline, for 120 min.day-1, 5 days.week-1. The rats were conditioned to run on a motor-driven treadmill by following a progressive two-week exercise program, so that by the end of the two weeks the rats were capable of running comfortably at this strenuous intensity in the non-pregnant state. Following the two-week running programme, the rats were paired by weight and randomly assigned to either a pregnant group that continued the running program throughout gestation (pregnant runner), or a pregnant group that did not continue the running program throughout pregnancy (pregnant control). At birth the neonates born to the pregnant running group did not differ in average neonatal body weight values, number per litter or total litter weight values when compared to controls, nor were superficial gross abnormalities observed in neonates born to the pregnant control or pregnant running groups. The strenuous maternal exercise intensity did not alter neonatal organ weight values (brain, heart, liver, lung, kidney), nor neonatal skeletal muscle (gastrocnemius, sternomastoid, diaphragm) when compared to control values. It is suggested that maternal exercise of this intensity throughout gestation does not affect fetal outcome in the rat, and may be due to the animals accustomization to the strenuous exercise protocol prior to pregnancy.     

A study of dose response and organ susceptibility of copper toxicity in a rat model

Copper (Cu) in higher concentration is toxic and results in various organ dysfunction. We report Cu concentration in liver, brain and kidney in the rat model following chronic exposure of oral copper sulphate at different subtoxic doses and correlate the tissue Cu concentrations with respective organ dysfunction. Fifty-four male wistar rats divided in 3 groups, the control group received saline water and the experimental group (Group-IIA and IIB) received oral copper sulphate in dose of 100 and 200 mg/kg Body Weight. At the end of 30 days, 60 days and 90 days of exposure, six rats were sacrificed from each group. The maximum peak force in grip strength, latency to fall in rotarod and percentage attention score in Y-maze were significantly reduced in the copper sulphate exposed rats compared to the controls at all time points and these were more marked in Group-IIB compared to Group-IIA. Cu concentration was significantly higher in liver, kidney and brain in the Group-II compared to the Group-I. The Cu concentration was highest in the liver (29 folds) followed by kidney (3 folds) and brain (1.5 folds). Serum ALT, AST and bilirubin correlated with liver Cu, BUN with kidney Cu, and grip strength, rotarod and Y-maze findings correlated with brain Cu level. In rats, chronic oral copper sulphate exposure at subtoxic level results in neurobehavioral abnormality and liver and kidney dysfunctions due to increased Cu concentration in the respective organs. Liver is the most vulnerable organ and copper toxicity increases with increasing dose and duration of exposure.     

DNA-protein cross-links in cells of internal organs of rats after fractional irradiation at various doses

The influence of daily fractional irradiation of male Wistar rats for 30 days on DNA-protein cross-links (DPC) in spleen, thymus, and liver cells was studied. The level of DPC depended strongly on the daily dose of irradiation and the studied organ. After irradiation at dose 0.5 Gy per day increased DPC level was detected in all organs. The highest level was in the lymphoid organs and the lowest in the liver. After irradiation at dose 0.3 Gy per day DPC formation was detected only in the thymus. The data suggest the existence of a dose threshold for DPC formation during fractional irradiation.     

Prolonged effect of stress (water and food deprivation) at weaning or in adult age on the triiodothyronine and histamine content of immune cells.

We used two days of total water and food deprivation as stress for female rats at weaning (three weeks old) and at adult age (two and a half months old). Triiodothyronine (T3) and histamine content of immune cells (lymphocytes, mast cells and monocyte-macrophage-granulocyte group in peritoneal fluid; lymphocytes, granulocytes and monocytes in blood; and lymphocytes in thymus) were studied three weeks after stress application using specific antibodies for flow cytometry and confocal microscopy. The stress at weaning increased T3 content of thymus lymphocytes. In case of adult T3, there was a cell type independent significant effect of stress, decreasing values in peritoneal fluid and slightly increasing effect in the blood. Histamine content of granulocytes was also significantly elevated. The experiments demonstrate that not only fetal or neonatal stress has long-lasting consequences, but also stress events in later periods of life in cells (organs) that are continuously differentiating. We will go on to discuss the importance of T3 and histamine in connection with stress and immunity.     

Effect of ovine follicular fluid peptide on ovarian responses and other organ weights in rats, Rattus norvegicus Berkenhout 1769

The present study was aimed to study the effect of an ovine follicular fluid peptide on ovarian follicle and good oocyte numbers and weights of ovary, uterus, liver, pancreas and kidney in rats, R. norvegicus. A 30.1 kDa peptide was isolated from ovine follicular fluid by ammonium sulphate precipitation and then gel filtration. The peptide was tested at various levels in normal (22 and 36 day-old), superovulated (29 day-old) immature and 121-day old mature rats on the ovarian responses and other organ weights. The isolated peptide inhibited the growth of antral follicles in normal and superovulated rats. Ovarian, uterine weight and recovery of good oocytes were reduced when the peptide was administered at 100 microg dose. The peptide had no effect on kidney, liver, pancreas weight and recovery of preantral follicles.