Modulation by neonatal thymectomy of the reproductive axis in male and female rats during development

The effects of neonatal thymectomy, at 3 days of age, on parameters of the reproductive axis were examined in male and female Sprague-Dawley rats. Gonadal and accessory sex tissue (male: epididymis, seminal vesicle, and ventral prostate; female: uterus) weights as well as anterior pituitary, spleen, and adrenal weights were determined in the thymectomized and sham-thymectomized animals at 20, 30, 40, 50, 60, and 90 days of age. Plasma gonadotropin concentrations as well as pituitary content of the gonadotropins and prolactin were assessed at each of these time intervals. No significant difference in gonad and accessory sex tissue weights was detected in thymectomized versus sham-operated controls at each of these times. Adrenal weights were increased in thymectomized animals compared with controls at 50 days of age and older in male rats and at 90 days in females. Spleen weights were decreased in the thymectomized males at 50 and 60 days of age. Thymectomy did not affect the spleen weight of females. Plasma concentrations of gonadotropins were unaffected in thymectomized males but were altered in females during the pre- and peripubertal period (Days 20-40). Vaginal opening, however, occurred at the same time in the thymectomized and control females. Pituitary gonadotropin and prolactin content were unaffected by thymectomy of the females, except at 90 days when pituitary luteinizing hormone (LH) content was lower in thymectomized than in control animals. LH and prolactin content were significantly reduced in the males at 60 and 90 days of age. These results demonstrate that there are sexual differences in the effects of thymectomy on parameters of the reproductive axis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of thymalin and heterotransfusion on blood coagulation and fibrinolysis in thymectomized rats

Experiments on rats have shown that thymectomy brings about the development of hypercoagulation and inhibition of fibrinolysis. Heterotransfusion is accompanied by hypocoagulation and stimulation of fibrinolysis in both intact and thymectomized rats. At the same time fibrinolysis in thymectomized rats is stimulated to a lesser degree than in intact animals. Preinjection into thymectomized rats of the thymus low-molecular factor thymaline over one week does not only make blood coagulation and fibrinolysis return to normal but also leads to adequate changes in the hemostatic system in response to heterotransfusion.     

Recapitulation of normal and abnormal BioBreeding rat T cell development in adult thymus organ culture

Congenitally lymphopenic diabetes-prone (DP) BioBreeding (BB) rats develop spontaneous T cell-dependent autoimmunity. Coisogenic diabetes-resistant (DR) BB rats are not lymphopenic and are free of spontaneous autoimmune disease, but become diabetic in response to depletion of RT6+ T cells. The basis for the predisposition to autoimmunity in BB rats is unknown. Abnormal T cell development in DP-BB rats can be detected intrathymically, and thymocytes from DR-BB rats adoptively transfer diabetes. The mechanisms underlying these T cell developmental abnormalities are not known. To study these processes, we established adult thymus organ cultures (ATOC). We report that cultured DR- and DP-BB rat thymi generate mature CD4 and CD8 single-positive cells with up-regulated TCRs. DR-BB rat cultures also generate T cells that express RT6. In contrast, DP-BB rat cultures generate fewer CD4+, CD8+, and RT6+ T cells. Analysis of the cells obtained from ATOC suggested that the failure of cultured DP-BB rat thymi to generate T cells with a mature phenotype is due in part to an increased rate of apoptosis. Consistent with this inference, we observed that addition of the general caspase inhibitor Z-VAD-FMK substantially increases the number of both mature and immature T cells produced by DP-BB rat ATOC. We conclude that cultured DR-BB and DP-BB rat thymi, respectively, recapitulate the normal and abnormal T cell developmental kinetics and phenotypes observed in these animals in vivo. Such cultures should facilitate identification of the underlying pathological processes that lead to immune dysfunction and autoimmunity in BB rats.     

The immunological role of the thymus in the pathogenesis of virus-induced lymphomas. Suppression of cytolytic T-cell function

Thymectomy of young adult mice has been found to prevent virus-induced lymphomas which develop as the animals age. Thymectomy protects mice by removing a source of suppressor T cells which inhibit the generation of cytolytic T cells against autochthonous tumors. Furthermore, suppression is specific since T cells are regulated in their capacity to respond to syngeneic but not allogeneic tumor cells. To determine if suppression could be adoptively transferred, lethally irradiated, bone-marrow-reconstituted mice were inoculated with T cells from either normal or thymectomized mice. Only T cells from thymectomized animals transferred enhanced T-cell reactivity to syngeneic tumor cells. More importantly, T cells from thymectomized mice injected with virus protected recipients challenged with lethal doses of syngeneic tumor cells. We conclude that thymectomy protects mice from developing virus-induced T-cell lymphomas by removing a source of suppressor T cells which regulates the activity of specific cytolytic T cells directed against autochthonous tumor cells.     

Thymic control of secretory antibody responses in the rat.

The effect of neonatal thymectomy on salivary and serum antibody responses was studied in rats. Local immunization of thymectomized rats with a T-dependent antigen (DNPBGG) elicited negligible amounts of IgA anti-DNP antibody in saliva. In contrast, both normal and sham-thymectomized animals demonstrated substantial levels of salivary IgA antibody. All thymectomized rats locally injected with a T-independent antigen (DNP-Lys-Ficoll) exhibited salivary IgA antibody production. Salivary IgG antibodies were somewhat decreased in thymectomized rats injected with either antigen; however, the final effect of T cell deprivation on IgG synthesis was not as pronounced as on IgA synthesis. Serum IgA antibody was induced in control rats injected with DNPBGG, whereas this Ig class of antibody was absent in thymectomized rats. The results suggest that thymus-derived cells exert a regulatory influence on both serum and secretory IgA responses to antigens.     

Control of IgE and IgG1 antibody production in mice.

The production of IgE and IgG1 was studied in untreated, thymectomized. splenectomized, anti-thymocyte serum-treated, or sublethally X-irradiated mice. Dinitrophenyl Ascaris and ovalbumin were used as antigens, and aluminum hydroxide was used as adjuvant. A suppression of IgE production was observed in adult thymectomized mice, although the kinetic pattern of the antibody response was the same as in control animals. IgG1 antibody production was not affected by thymectomy. Splenectomy did not change either IgE or IgG1 production. A single dose of rabbit anti-thymocyte serum (ATS) given 8 days after immunization inhibited IgE antibody production. The effect of ATS was dose dependent and also varied with the amount of antigen used, the immune response to high doses being more susceptible to the effect of ATS. No alteration in IgG1 production was caused by ATS even when IgE antibody formation was completely inhibited. When preceding immunization, sublethal irradiation enhanced IgE antibody formation and partially suppressed IgG1 production; applied after immunization, irradiation caused an enhancement of IgE production which was inversely proportional to the interval elapsed between the two procedures. On the other hand, the IgG1 antibody production was fairly resistant to the same treatment. The results suggest a clearcut separation between the mechanisms regulating IgE and IgG1 production in mice.     

Lymphoid organ development in Xenopus thymectomized at eight days of age

This paper examines the effect of early thymectomy on the subsequent development of lymphoid tissues in the toad, Xenopus laevis. At the time of thymic removal (8 days post-fertilization) all the lymphoid organ anlagen are at a rudimentary state of differentiation and contain few, if any, small lymphocytes. Despite the absence of any thymic tissue all thymectomized animals grew normally. Thymectomized larvae developed relatively normal lymphoid organs. However, lymphoid depletion was apparent in the splenic red pulp and in the pharyngeal ventral cavity bodies. Examination of the lymphoid organs of post-metamorphic Xenopus revealed reduction in spleen size following thymectomy. Lymphoid depletion was evident in the splenic red pulp of many thymectomized toadlets and reduction in proportion of white to red pulp was also noted in a few of these animals. Absence of the thymus had no apparent effect on the histology of the other lymphoid organs examined.     

Effect of antilymphocyte serum and neonatal thymectomy upon onset of diabetes in the Chinese hamster

Prediabetic Chinese hamsters were treated with antilymphocyte serum (ALS), or thymectomized in order to test the hypothesis that beta-cell loss leading to diabetes in this animal model was related to cell-mediated autoimmunity. In addition, passive transfer of diabetes from the Chinese hamster to the nude mouse was attempted by transplantation of lymphocytes. Treatment of prediabetic Chinese hamsters with ALS or thymectomy did not alter development or severity of diabetes in this animal model. Lymphocytes from newly diagnosed diabetic Chinese hamsters did not cause hyperglycemia in nude mice. These three lines of evidence suggest that cell-mediated autoimmunity does not contribute to the etiology of diabetes in the Chinese hamster. The Chinese hamster remains a good model for the study of those forms of diabetes not related to cell-mediated autoimmune phenomena.     

Lymphocyte-mediated cytotoxicity: effects of ageing, adult thymectomy and thymic factor.

Adult thymectomy, as well as ageing, depressed splenic lymphocyte-mediated cytotoxicity (LMC) in the mouse. Ageing depressed significantly LMC as early as 19 weeks of age, independently of the number of cells used for immunization. Thymectomy affected LMC only when supoptimal numbers of immunizing allogeneic cells were used. This effect peaked at 6 to 12 weeks after thymectomy. No difference between thymectomized and normal mice was observed when LMC was tested 16 to 20 weeks after thymectomy, at an age when normal control mice themselves already showed a lowered LMC due to ageing. The effect of in vivo treatment with a circulating thymic factor (TF), which was shown to disappear with ageing as well as after adult thymectomy, has been tested in adult thymectomized mice and normal young and ageing mice. TF treatment prevented LMC depression in adult thymectomized mice, whereas it depressed paradoxically splenic LMC in normal young and old mice. The possible mechanisms of the effects of adult thymectomy, ageing, and thymic factor on the different T cell subsets involved in allogeneic killer cell generation are discussed.     

Pup Shoving by Adult Naked Mole-Rats

Adult naked mole‐rats (Heterocephalus glaber) characteristically perform an unusual behavior toward young: they shove small pups frequently and vigorously around the nest. We studied 15 litters in five captive colonies to quantify which adults shove pups, changes in shoving frequencies as pups develop, how external disturbances affect pup‐shoving frequencies, and behavior of juveniles that were not shoved as pups. In all litters and colonies the breeding female shoved pups significantly more often than any other individual. Breeding females also shoved adult colony mates, but at far lower rates than they shoved pups. Breeding males shoved pups about half as often as did breeding females. Together, the parents shoved pups ten times more often than did nonbreeders. Frequencies of pup shoving peaked when pups were 3–4 wk old, roughly coincident with weaning. When colonies were disturbed experimentally, frequencies of pup shoving increased dramatically, whereas rates at which nonbreeding adults were shoved decreased sharply. We separated four newly‐weaned litters and raised half the pups apart from their colony. When these litters were reunited 4–9 wk later, the unshoved (experimental) pups were the same size as the frequently shoved (control) pups, but the unshoved pups were significantly less likely to flee from a disturbance. Shoving of small pups encourages them to flee from danger, and also may enforce weaning.     

Breeding colony refinement through phenotypic and genotypic characterization of the SPRD-Pkdr1/Rrrc rat model of polycystic kidney disease

The SPRD-Pkdr1 rat model is widely used for the study of human autosomal dominant polycystic kidney disease. This rat model carries the Cy allele of the Pkdr1 gene, which results in polycystic kidney disease. Because the Cy allele is lethal in the homozygous state at weanling age, the breeding colony must be maintained in the heterozygous state. A random breeding scheme in which production of homozygous pups with enlarged kidneys indicates heterozygous breeders is commonly used. This study was performed to determine whether biochemical markers (blood urea nitrogen [BUN] or creatinine), ultrasonography, or genetic analysis could be used to select breeding animals in the SPRD-Pkdr1/Rrrc colony and thus replace the random breeding scheme with a more efficient selective breeding scheme. BUN was predictive of the Cy allele in 8- to 9-wk-old male but not female rats. Ultrasonography identified animals with polycystic kidney disease in both sexes by 9 wk of age. Microsatellite marker polymorphism analysis could not be used to determine carrier status for the Cy allele, but restriction fragment length polymorphism analysis appropriately detected the Cy allele in 100% of the animals examined. In conclusion, multiple methods can be used for detecting the Cy allele, making possible a selective breeding scheme that markedly reduces the necessary number of breeder animals and eliminates the euthanasia of offspring needed with a random test-mating scheme.     

Neonatal thymectomy affects follicle populations before the onset of autoimmune oophoritis in B6A mice

Using a variation on a standard follicle classification technique, 5 classes of follicles were quantified in serial sections of ovaries from intact mice and mice thymectomized on Day 3 at 5-day intervals from 5 to 40 days of age. Sera from these animals and from animals 60 days of age were analysed for the presence of anti-oocyte antibodies. Ovaries from intact animals 10 to 40 days of age were examined for the presence of antigen(s) using anti-oocyte antibody-positive sera from all ages of mice. There was a dramatic decrease in the primordial follicle population at 10 days of age in thymectomized mice and that population remained significantly lower until 40 days of age. The growing follicle population was also significantly lower at 20 days of age in thymectomized mice and remained lower through 40 days of age. Anti-oocyte antibodies were not detectable until 30 days of age and at that age reacted with oocytes from all follicle types including primordial. Ovarian antigens were present in similar patterns in ovaries from mice at all ages tested. We conclude that thymectomy has an earlier influence on the ovary than previously thought and this influence does not appear to involve the immune activity associated with autoimmune ovarian dysgenesis. This suggests that the effect of thymectomy on the ovary may be biphasic: (1) an early effect, possibly involving a disruption in the hypothalamic-pituitary-ovarian-thymic axis, that influences the primordial and growing follicle populations before 20 days of age; and (2) a later effect involving an immune imbalance first evident by 25-30 days of age that ultimately results in the destruction of the ovary.     

Anti-immunoglobulin stimulation of murine lymphocytes. III. Enhancement of the development of responsive B cells by thymic deprivation.

The development of splenic B cells that can be induced to proliferate by soluble anti-immunoglobulin (anti-Ig) reagents requires 7 to 9 months in normal mice. We have found that this age-associated response is enhanced by thymic deprivation. Both neonatally thymectomized LAF₁ mice and thymectomized, lethally irradiated, and bone marrow-restored Balb/c mice respond earlier and more strongly to anti-Ig than their sham controls. Nevertheless, at least 3–4 months are still required after thymectomy before a response can be measured. The earlier and enhanced response to anti-Ig seen in thymectomized animals is not due simply to an increase in the total number of Ig-positive spleen cells. The age-associated response of splenic B cells to anti-Ig we have observed in normal mice may be explained by the “natural” loss of thymic influence that occurs with age.     

In vitro responses of rat lymphocytes following adult thymectomy. II. Increased inhibition by splenic adherent cells of responses to phytohemagglutinin

Adult thymectomy in rats results in a marked fall of spleen cell responsiveness to PHA over a period of days to weeks. Examination of dose-response curves showed that, with high PHA dose or ≥ 10⁶ cells/ml, there is a profound inhibition of the response with spleen cells from thymectomized animals compared with cells of matched sham-operated controls. However, when adherent cells are removed from the cell suspensions, the remaining nonadherent cells give an almost linear dose-response relationship with increasing PHA similar to that exhibited by the nonadherent cells of the controls or sometimes a slightly decreased response. Similarly, when increasing numbers of spleen cells from these animals are cultured (with admixed thymocytes to make a constant total of 2 × 10⁶ cells/ml) with PHA, the linear portion of the doseresponse curve can be extrapolated to give a similar value for the maximal potential response, which again is the same as or somewhat less than the corresponding value for sham-operated controls. A difference in inhibitory capacity is also shown in mixtures of the two spleen cell populations with LNC or with purified spleen cells. It is concluded that adult thymectomy results in increased “suppressor” activity in the spleen within a few days and may reduce slightly the number of T lymphocytes in the spleen reactive with PHA.     

Reproduction and survival of suricates (Suricata suricatta) in the southern Kalahari

In most respects, the demography of Kalahari suricates (Suricata suricatta) resembles that of other social mongooses. Average group size varies from four to nine, and groups typically include several mature females, of which one is responsible for the majority of breeding attempts. Breeding females show a postpartum oestrus; gestation is around 60 days; litter size is three to five pups at emergence and females rarely breed before the age of 24 months. In contrast, annual survival rates (0.20 for pups and 0.43 for animals over one year old) are lower than those recorded in other species. Breeding frequency is related to rainfall and breeding can cease altogether when rainfall is unusually low. In a year when this occurred, group size eroded rapidly and over 60% of groups became extinct. Total numbers were slow to recover during the following year because emigration by females was infrequent and new groups did not form in vacant ranges created by the extinction of groups. High rates of group extinction have been found in other cooperative breeders and may occur because breeding success and survival show inverse density dependence.     

Development of echolocation and communication vocalizations in the big brown bat, <Emphasis Type="Italic">Eptesicus fuscus</Emphasis>

Big brown bats form large maternity colonies of up to 200 mothers and their pups. If pups are separated from their mothers, they can locate each other using vocalizations. The goal of this study was to systematically characterize the development of echolocation and communication calls from birth through adulthood to determine whether they develop from a common precursor at the same or different rates, or whether both types are present initially. Three females and their six pups were isolated from our captive breeding colony. We recorded vocal activity from postnatal day 1 to 35, both when the pups were isolated and when they were reunited with their mothers. At birth, pups exclusively emitted isolation calls, with a fundamental frequency range <20 kHz, and duration >30 ms. By the middle of week 1, different types of vocalizations began to emerge. Starting in week 2, pups in the presence of their mothers emitted sounds that resembled adult communication vocalizations, with a lower frequency range and longer durations than isolation calls or echolocation signals. During weeks 2 and 3, these vocalizations were extremely heterogeneous, suggesting that the pups went through a babbling stage before establishing a repertoire of stereotyped adult vocalizations around week 4. By week 4, vocalizations emitted when pups were alone were identical to adult echolocation signals. Echolocation and communication signals both appear to develop from the isolation call, diverging during week 2 and continuing to develop at different rates for several weeks until the adult vocal repertoire is established.     

Trypanosoma cruzi: effects of anti-thymocyte serum in mice and neonatal thymectomy in rats

CF₁ mice were given eight injections of normal rabbit serum (NRS), Hanks' balanced salt solution (HBSS), or rabbit anti-mouse thymocyte serum (ATS) beginning 3 days prior to and at 3-day intervals subsequent to intraperitoneal (ip) inoculation with 5 × 10⁴ trypomastigotes of a Brazil strain of Trypanosoma cruzi. Markedly enhanced parasitemia, increased numbers of tissue stages (amastigotes), and higher mortality occurred in ATS-treated mice as compared to NRS- or HBSS-treated controls. Administration of three injections of ATS at 3-day intervals during the latter stages of acute Chagas' disease, i.e., when numbers of parasites were declining, resulted in a transitory relapse (increase in numbers) of blood and tissue parasites. No relapse occurred in mice when ATS was administered at 3-day intervals over a period of 15 days during the subacute stage of the disease, i.e., after parasites had disappeared from the blood.Parasitemia and mortality were enhanced in neonatally thymectomized rats when compared to that observed in sham-operated and unoperated control rats following ip injection of 2 × 10⁵ trypomastigotes of T. cruzi. Serum obtained from thymectomized and control rats 5 weeks after inoculation with T. cruzi at a time when the blood of all animals had become microscopically negative for parasites were equally protective in passive transfer experiments, while serum from uninfected controls gave no protection.Gamma globulin levels significantly increased in thymectomized as well as intact rats by the third to fourth week of infection with T. cruzi, reached maximum concentrations in 5–6 wk, and remained elevated significantly at the twelfth week post infection as compared with uninfected controls. No significant changes occurred in total serum proteins or α and β fractions of any group, infected or uninfected.Total circulating leukocytes, especially lymphocytes, were diminished in mice and rats subjected to treatment with ATS or neonatal thymectomy.These data clearly indicate that neonatal thymectomy of rats and ATS treatment of mice suppress the acquired immune response to T. cruzi. Further, passive transfer experiments in rats confirm the protective role of circulating antibody in acquired immunity to Chagas' disease.     

胸腺对雌性大鼠肝脏抗氧化功能的影响及其与细胞免疫、性激素水平的关系

雌性成年大鼠切除胸腺后,其肝脏脂质过氧化产物丙二醛增多,还原型谷胱甘肽(GSH)含量减少,微粒体和线粒体膜流动性降低。隔日1次皮下注射自猪胸腺提取的胸腺因子D(TFD)2mg/kg共3月,可逆转去胸腺大鼠上述指标的变化。为了探讨胸腺影响肝脏抗氧化功能的中间途径,还测定了脾T淋巴细胞增殖反应及血浆性激素水平。结果表明,去胸腺大鼠脾脏T淋巴细胞增殖反应减弱、血浆雌二醇/睾酮比值下降;给予TFD可逆转其变化。实验结果提示,胸腺对大鼠肝脏抗氧化功能的影响可能与胸腺-内分泌-免疫网络有关。     

Lack of effect of vitamin D administration during pregnancy and early life on diabetes incidence in the non-obese diabetic mouse.

BACKGROUND AND AIMS: Several studies have suggested that vitamin D supplementation in early life may reduce the risk of developing type 1 diabetes in later life. The non-obese diabetic (NOD) mouse is a model of spontaneous type 1 diabetes currently used for testing hypothesis/compounds aimed at disease prevention. In this study, we tested the effect of vitamin D (16 IU by gavage) on diabetes incidence in NOD/Ba mice treated from conception with olive oil containing vitamin D via maternal dosing up to 10 weeks of age and followed up until 32 weeks of age. METHODS: Twelve breeding pairs were administered olive oil containing vitamin D during pregnancy, 15 days following the birth of the pups and for the next 10 weeks subsequently. The same breeding pairs were bred again after a clearance period of 15 days using a control solution to produce a control litter. This control group received a control solution for the same period of time. Diabetes incidence, degree of insulitis, and insulin content in the pancreas were investigated in the two groups. RESULTS: 12 vitamin D-treated NOD mice developed diabetes compared to 15 animals in the control group (Log rank test p = 0.899, NS). There were no significant differences between the groups in diabetes incidence, time of onset of the disease, degree of insulitis, or the insulin content in the pancreas. CONCLUSION: Vitamin D administered in utero and in the early stages of life at the dosage used does not change the incidence of diabetes or modify the disease process that leads to beta cell destruction in the NOD mouse.     

The influence of sialoadenectomy, thymectomy and starvation on liver glycogen in the rat

Hepatic glycogen was assayed in young and adult rats subjected to sialoadenectomy and/or thymectomy and starvation. Sialoadenectomy in young, but not in adult rats caused the rats to stop feeding. In young, but not in adult sialoadenectomized and starved rats the glycogen level was notably higher than in unoperated and starved rats, indicating active participation of salivary glucagon in immature animals in hepatic glycogenolysis under conditions of starvation. Simultaneous sialoadenectomy and thymectomy caused glycogen depletion in the liver of young rats in spite of the absence of the salivary glands. Acceleration of glycogenolysis in these rats was not due to thymectomy, being probably a result of excessive secretion of adrenal catecholamines.