Chronic alpha1-adrenoreceptor blockade produces age-dependent changes in rat thymus structure and thymocyte differentiation

In order to examine the influence of chronic alpha1-adrenergic receptor (alpha1-AR) blockade on the thymus structure and T-cell maturation, peripubertal and adult male rats were treated with urapidil (0.20 mg/kg BW/d; s.c.) over 15 consecutive days. Thymic structure and phenotypic characteristics of the thymocytes were assessed by stereological and flow cytometry analysis, respectively. In immature rats, treatment with urapidil reduced the body weight gain and, affecting the volume of cortical compartment and its cellularity decreased the organ size and the total number of thymocytes compared to age-matched saline-injected controls. The percentage of CD4+8- single positive (SP) thymocytes was decreased, while that of CD4-8+ was increased suggesting, most likely, a disregulation in final steps of the positively selected cells maturation. However, alpha1-AR blockade in adult rats increased the thymus weight as a consequence of increase in the cortical size and cellularity. The increased percentage of most immature CD4-8- double negative (DN) cells associated with decreased percentage of immature CD4+8+ double positive (DP) thymocytes suggests a decelerated transition from DN to DP stage of T-cell development. As in immature rats, the treatment in adult rats evoked changes in the relative numbers of SP cells, but contrary to immature animals, favoring the maturation of CD4+8- over CD4-8+ thymocytes. These results demonstrate that: i) chronic blockade of alpha1-ARs affects both the thymus structure and thymocyte differentiation, ii) these effects are age-dependent, pointing out to pharmacological manipulation of alpha1-AR-mediated signaling as potential means for modulation of the intrathymic T-cell maturation.     

Age related changes in total DNA and RNA and incorporation of uridine and thymidine in rat brain

1. Total brain DNA and total brain RNA and the incorporation of thymidine[14C] and uridine[3H] were measured in young and aged rats. 2. From 20 days to the time of sexual maturation, both DNA and RNA levels increase. Total RNA exceeds total DNA at all ages. Comparatively, the ratio of total DNA/RNA is higher in young than in aged animals. 3. The incorporation of thymidine[14C]/g of DNA and of uridine[3H]/g of RNA decreases with age. This decrease is rapid in young animals. After 350 days of age, the incorporation becomes very low. The significance of data is discussed.     

Cellular distribution, developmental changes and effects of cryptorchidism on uridine kinase in the rat testis

High specific activity of uridine kinase was found in cultured peritubular cells (3.0 nmol/min per mg protein) which was more than 3-fold higher than that found in cultured Sertoli cells (0.79 nmol/min per mg protein). In the various classes of germ cells a decrease in specific uridine kinase activity was associated with increased maturity of the cells, primary spermatocytes, round spermatids and spermatozoa showing 1.3, 0.65 and 0.16 nmol/min per mg protein, respectively. A relationship between uridine kinase activity and the rate of RNA synthesis in these cells is suggested. A decrease in specific uridine kinase activity in testis with increasing age supports the finding of lower uridine kinase in mature germ cells than in earlier germ cells and somatic cells. This finding is further supported by the observation that cryptorchidism, which is associated with a time-dependent depletion of germ cells, resulted in an increase in specific uridine kinase activity. The results indicate that pyrimidine salvage is important in earlier germ cells, as well as in somatic cells in the testis, to produce substrates for nucleic acid synthesis.     

Immunological changes in the thymus epithelial tissue of mice during accidental involution

Using sera containing antibodies to antigens of epithelial reticular cells it is possible to detect by immunofluorescence early stages of organ accidental involution and to trace parenchyma demasking that results from lymphocyte migration upon injection of prednisolone, azathioprin or their combinations. Prior to the drug administration only certain short thin processes of epithelial cells masked by a great number of lymphoid elements are detectable in the cortical zone of the thymus. Injection of 10-40 mg/kg prednisolone, 100-200 mg/kg azathioprin or their combinations causes stable different-degree demasking of the epithelial reticulum seen during immunofluorescence as a network of fluorescent processes. Administration of drug combinations also causes marked attenuation of the reaction, probably resulting from dystrophic changes in thymic epithelial cells.     

The postpartum buffalo: I. Endocrinological changes and uterine involution

To maintain a calving interval of 13-14 months in buffaloes, successful breeding must take place within 85-115 days after calving. Disturbances during this period due to delay of uterine involution or resumption of estrous activity are likely to prolong the calving interval and reduce the lifetime reproductive and productive efficiency. In this article literature on endocrinological changes in the peripartum period and on factors affecting uterine involution are reviewed. The available information indicated that although the availability of releasable FSH does not appear to be a limiting factor for resumption of postpartum cyclicity a substantial increase of releasable LH and replenishment of pituitary stores occurred around Day 20 in dairy and Day 30 in swamp buffaloes. There is evidence that follicular activity is resumed early (15-30 days) in the postpartum period. However, the factors which initiate release of appropriate LH pulses, follicular maturation and ovulation in the postpartum buffalo need further studies. The mean interval to complete uterine involution varied widely between 19 and 52 days. Assessment of cervical and uterine horn diameters by rectal palpation alone is not satisfactory to diagnose delayed uterine involution and possible subclinical uterine infection. Vaginal inspection can be included as a fundamental part of postpartum genital examination for diagnosis of such case. Uterine involution, however, does not seem to be a limiting factor for achievement of satisfactory fertility in the postpartum buffalo but the main determinant is resumption of estrous activity.     

Age-related changes in total dna and rna and incorporation of uridine and thymidine in rat liver,kidney and spleen

• 1.1. Total content of DNA and RNA in liver, kidney and spleen were measured in young and aged rats. At the same time the incorporation of [¹⁴C]thymidine, a DNA precursor, and [³H]uridine, an RNA precursor, were also determined.
• 2.2. Changes in total organ DNA and RNA correlated with sexual maturation as did incorporation of precursors.
• 3.3. Young animals have more DNA per organ relative to RNA. with kidney and spleen DNA showing a decrease between maturity and senescence.
• 4.4. However, liver RNA increases with age. a change probably due to decreased catabolism of RNA since [³H]uridine uptake decreases.
• 5.5. Liver polyploid differentiation, and [¹⁴C]thymidine and [³H]uridine uptake, are correlated.
• 6.6. In kidney, incorporation of [³H]uridine is inversely related to [¹⁴C]thymidine incorporation.

     

Density-dependent cell division after cortisol treatment of rat thymus in relation to age involution.

Cellular proliferation, in relation to cell density was investigated in the thymus of control and cortisol treated animals at 6 and 18 weeks of age. It was found that there was very little difference in the response of the two age groups to cortisol treatment. Cell density and cellular proliferation were markedly reduced 2 days after cortisol administration. From 4 days there was a rapid increase in cellular proliferation to triple the control rate. The mitotic index remained above normal until 12days then decreased to control values at 14 days. During this time the cell density of the thymus was being progressively restored. At all stages of regeneration, the mitotic index at first increased to a maximum at the mean cell density then decreased at the highest cell concentrations. A model system is discussed to account for this density dependent control of cellular proliferation in the thymus.     

Vascular and stromal changes in irradiated and recovering rat thymus.

To analyze the mechanisms responsible for thymocyte proliferation, maturation and migration in the thymus, the rat thymus just after, and recovering from irradiation was studied morphologically. The vascular structures of the rat thymus after a radiation dose of 6 Gy were found to be destroyed on day 3, but had recovered to almost normal by day 7, suggesting that the abrupt recovery of thymus structure after irradiation was due primarily to this change in vascular structure. Furthermore, the epithelial tissues in the thymic cortex appeared to contribute to this abrupt proliferation, and possibly to the abrupt maturation of thymocytes, while medullary epithelial tissues remained sparse and appeared inactive for a relatively long period. These findings are considered important for understanding the interrelationship between thymic epithelial cells and thymocytes with respect to thymocyte proliferation, maturation and migration.     

Cellular localization and age-dependent changes in mRNA for cyclic adenosine 3',5'-monophosphate-dependent protein kinases in rat testis

Gonadotropin activation of cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinases plays an important role in the regulation of testicular function. This study was undertaken to establish the expression of various subunits of cAMP-dependent protein kinases in different testicular cell types as well as during sexual maturation. RNA was extracted from cultured Sertoli cells, cultured peritubular cells, germ cells (pachytene spermatocytes, round spermatids), tumor Leydig cells, as well as whole testis from rats of various ages. Messenger RNA levels were studied by Northern analysis using available cDNA probes. The regulatory subunit (R) designated RII51 was found to be predominantly expressed in cAMP-stimulated Sertoli cells and tumor Leydig cells. Much lower levels were found in cultured peritubular cells and germ cells. A 2.9- and 3.2-kb mRNA for the RI subunit were found at about similar levels in all cell types, whereas the smaller 1.7-kb mRNA was expressed in high levels in germ cells. Also, the catalytic subunit (C) of cAMP-dependent protein kinase, designated C alpha, was expressed in all cell types; the highest mRNA levels for this subunit were found in germ cells and in tumor Leydig cells. The 1.7-kb mRNA for androgen-binding protein (ABP) was abundant in cAMP-stimulated Sertoli cells and was not present in other cell types of the testis. Furthermore, the cellular localization of the cAMP-dependent protein kinase subunits was also supported by developmental studies. The mRNA level of the RII51 3.2-kb species was relatively constant until Day 30, after which there was a tendency to decrease. A 1.6-kb message first appeared at greater ages. The mRNA for the smaller 1.7-kb species of RI, as well as the C alpha, showed a significant increase during development, supporting an enrichment of these mRNAs in germ cells. Messenger RNA levels for ABP were not detected in testis from 5- to 10-day-old rats but increased up to Day 30. After this age, mRNA for ABP revealed an age-dependent decrease, which parallels the relative increase of germ cells in the testis. In summary, these results demonstrate a clear pattern of cellular localization of the various mRNA species for subunits of the cAMP-dependent protein kinase in the rat testis.     

Concanavalin A-induced chemiluminescence in rat thymus lymphocytes. Its origin and role in mitogenesis.

Incubation of a particulate preparation from potato tissue culture cells with UDP-beta-L-[1-3H] arabinose yielded a glycoprotein fraction containing labelled material with the characteristics of hydroxyproline arabinosides. The sugar-protein linkage was resistant to hot alkaline hydrolysis, and the hydrolytic products showed similar electrophoretic and chromatographic behavior to authentic hydroxyproline-arabinosides prepared from potato tissue culture cell walls. Incorporation of arabinose into glycoprotein was stimulated by the addition of de-arabinosylated potato lectin. The product of the incubation co-migrated with native potato lectin on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The subcellular distribution of the arabinosyl-transferase was investigated by fractionating potato tissue culture membranes on a discontinuous sucrose gradient in the presence or absence of Mg2+. Under both fractionation conditions the highest specific activity of the enzyme was found in the Golgi-enriched fraction. The results are discussed in relation to the synthesis of the hydroxy-proline-rich glycoprotein component of plant cell walls.     

Cellular distribution of prothymosin alpha and parathymosin in rat thymus and spleen

By means of immunohistochemical methods, we have investigated the cellular distribution of prothymosin alpha and parathymosin in rat thymus and spleen, using specific antibodies raised against thymosin alpha-1 and against parathymosin. We observed prothymosin alpha immunoreactivity in lymphoid cells both in thymus and spleen. In the thymus, prothymosin alpha staining was more marked in cortex than in medulla. In the spleen, prothymosin alpha was found in lymphocytes of the periarteriolar lymphatic sheaths and was especially prominent in the germinal centers. Parathymosin immunoreactivity in the thymus was mainly localized in the medulla; positive cells were reticuloepithelial cells from the thymic reticulum and the blood barrier. Thymocytes were negative. In spleen, parathymosin was found in reticular cells arranged in a ring between the periarteriolar lymphatic sheath and the marginal zone. Our results do not support an exclusive role for these peptides as immune system hormones or cytokines.     

Antigenic markers on rat lymphocytes. I. Characterization of A.R.T., an alloantigenic marker on rat thymus and thymus-derived cells.

Drugs with pharmacologic activities on cell membrane components were tested for their effects on responses of murine lymphocytes to mitogens. The effects of the drugs were found to be partly selective and related to the type and dose of the mitogen. Most striking were the effects of cytochalasin B, which inhibited the responses to low doses of concanavalin A or phytohemagglutinin, but potentiated the reactions to the high doses of the lectins. Chloroquine and chlorpromazine, on the other hand, were more inhibitory to cultures containing supraoptimal doses of the lectins. Colchicine inhibited the responses against lipopolysaccharide more than those against the two lectins. The inhibitory effects of colchicine were almost unchanged when the drug was added at intervals up to 24 hr after the onset of culture, whereas the effects of the other tested drugs diminished markedly when added to cultures 6 hr or more after the mitogens. The results are discussed in view of their relationship to the poorly understood mechanisms which regulate the lymphocyte responses in the presence of different doses of mitogens.     

Evolution, development and involution of the thymus

We studied the appearance of the thymus primordia of main representatives of the vertebrate evolution, observed these anlages in different places of the branchial region and followed their gradual reduction in phyletic line. On the assumption that the thymus potency has been encoded in evolution, we studied the human thymus ontogeny and described the entodermal thymus primordia not only in the 3rd but also in the 2nd and 4th pouch. We describe a very important contribution of the ectodermal canaliculus, projecting from the 3rd cleft, as well as the participation of the hypobranchial neural placode to the definitive thymus formation. Even though the entodermal thymus primordium in the 2nd pouch has reached an advanced stage, the thymus cannot complete its development because the ento-ectodermal relation is missing. In addition to this dysgenesis, we describe thymic involution in a later fetal period in connection with some malformations and confirm the close relation between the central immune organ and the neuro-endocrine system.     

Effect of T-activin on thymus involution in mice

The effect of T-activin on thymus involution in mice was studied. T-activin in a dose of 1.0 micrograms/mouse was injected into young male (CBA X C57BL)F1 mice weighing 24.0 +/- 2.0 g daily for 20 days. Morphometric analysis of the thymus was made 6 months after the treatment with T-activin was completed. It was found that T-activin induced the suppression of physiological involution of the thymus together with the enhancement of the processes of thymocyte transformation and proliferation.