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1.
A 72-hour treatment of Tetrahymena with serotonin analogues at a concentration of 10(-9) mmole/l resulted in a decrease of the reproduction rate, whereas serotonin itself was ineffective. On the second exposure to antagonists, cellular division was enhanced but its rate remained around or below the control value, while the second exposure to serotonin produced a more marked effect than the first one did, which finding implies a receptor amplification. The most pronounced receptor amplification was caused by the first exposure to serotonin. This observation indicates that the receptor is rather selective as regards the amplification effect.  相似文献   

2.
It is known from model experiments on Tetrahymena that primary exposure to a hormone induces receptor formation or amplification, in other words a hormonal imprinting. Substances acting on the intracellular Ca2+ level of the Tetrahymena, such as TMB-8, EDTA, EGTA, NiCl2 and La(NO3)3, interfered with hormonal imprinting of the unicellular to different degrees, and some of them influenced hormone (insulin, TSH) binding also independently of imprinting. Interference with the intracellular Ca-metabolism generally influenced imprinting by insulin and TSH, which were mediated by different mechanisms, to dissimilar degrees, or in opposite directions. On combined application of the agents acting on Ca-metabolism, their effects were additive. It appears that intact Ca-mediation is an essential prerequisite for normal hormonal imprinting.  相似文献   

3.
Chemotactic selection is a method by which populations of cells exposed to ligands can be isolated and subsequently cultivated. We used Tetrahymena pyriformis GL cultures selected by chemotactic selection to insulin (10 nM), histamine (0.1 nM) and di-iodotyrosine (T2, 10 nM) to study the phagocytotic capacity under the induction of selector hormones. Our results show a long-lasting link between chemotactically selected cultures and phagocytotic activity. Cells selected to histamine produced the highest phagocytotic activity upon a second exposure to the selector hormone. T2 selection was also strongly effective, however, the phagocytosis stimulation was not specific to the hormone given later. Insulin selected sub-populations had different phagocytotic responses to the control substance itself, whereas histamine selected sub-populations seem to be heterogeneous in the phagocytotic response to histamine. For insulin, the increased endocytotic or metabolic activity was demonstrated by the lack of non-phagocytotic cells. These experiments call attention to the evolutionary role of selection in the later developing receptor-hormone relationship.  相似文献   

4.
The hormone combination epinephrine + diiodotyrosine inhibited the growth of Tetrahymena cells on first exposure, but stimulated it markedly on re-exposure. It appears that amplification of the receptor by the hormone does take place at the first encounter, regardless of whether the response of the cells was positive or negative. The amplifying effect persists over several generations. The intensity of stimulation by the second exposure was directly related with the duration of the first one. Prolongation of the first exposure accounted for a switch-over from negative to positive influence.  相似文献   

5.
1. Treatment of Tetrahymena pyriformis cells with diiodotyrosine (T2) gave rise to a considerable, concentration-dependent increase of the growth rate within the range of 10(-15) and 10(-9) M, but did not influence it at the level of 10(-18) M. 2. Re-exposure of the cells 1, 2 and 4 weeks later to the hormone concentrations originally used accounted for a marked increase of growth rate at all hormone levels tested, indicating that the extremely low concentration of 10(-18) M, which failed to stimulate growth on first exposure, did nevertheless give rise to hormonal imprinting, which caused the cells to "remember" the hormone, as judged from their increased responsiveness to it on re-exposure. 3. The degree of growth response was concentration-dependent on both first and second exposure: higher levels of treatment gave rise to firmer imprinting, and to greater response on re-exposure. 4. The length of exposure time proved to be more decisive than the level of treatment in respect of the development of hormonal imprinting. 5. Short-term exposures up to 60 min, although they stimulated cell growth by direct effect, gave rise to lasting inhibition of cellular response to re-exposure(s) rather than to hormonal imprinting.  相似文献   

6.
The first interaction between target cell and hormone gives rise to hormonal imprinting, which accounts for greater responsiveness of the cell at later interactions. The mechanism of hormonal imprinting is obscure; we based experimental approach to its closer study on combined treatment of Tetrahymena, as model cells, with diiodotyrosine (T2), which stimulates the division, and cell growth inhibitors, which interfere with different stages of cell reproduction, and methylamine, which inhibits cluster formation in the membrane. Of these, actinomycin D and methylamine inhibited the growth of the Tetrahymena, while colchicine did not, and all three suppressed the division stimulating action of T2, but could not prevent hormonal imprinting, as demonstrated on later re-exposure to T2 of cells preexposed and not preexposed to T2 in combination with the inhibitors. It appears that the underlying mechanism of hormonal imprinting is highly complex, and involves many subcellular mechanisms and structures, but suppression of, or gross interference with, one or another of these cannot delete, only quantitatively reduce, the consequence of the first interaction with the hormone, i.e. hormonal imprinting.  相似文献   

7.
Functional overlap of FSH and TSH in cockerel testicle is expressed by alteration of testicular weight and canalicular diameter length. While on neonatal exposure TSH was more active than FSH, at five weeks of age both hormones developed an equal action. A single neonatal exposure to FSH had a durable effect on testicle weight and canalicular diameter, while the effect of post-hatching TSH persisted only in respect of the second parameter. A single neonatal exposure to TSH failed to amplify TSH effect on a second exposure at five weeks of age, while neonatal TSH and FSH treat equally enhanced the effect of FSH given at five weeks. This suggests that under given conditions, perinatal hormone excess may account for lasting amplification of the receptor(s) not only in respect of the adequate hormone, but also in respect of a related hormone, hormone-like material or analogon.  相似文献   

8.
Diiodotyrosine and serotonin enhance the growth of Tetrahymena. A second exposure of the unicellular to these hormones accounts for a still greater increase of its growth rate, probably due to the amplification of the receptor induced by the first exposure.  相似文献   

9.
The aim of the experiments was to study the regulation of triiodothyronine (T3) production in the unicellular Tetrahymena. Untreated and troph-hormone treated specimen were prepared and in different timepoints T3 content was measured and compared by immunocytochemical flow cytometry. 0.1 or 0.001 IU TSH in tryptone-yeast medium stimulated T3 synthesis at 10, 20, 30 min, but does not stimulate after 1 h. The overlapping gonadotropic hormone (GTH) also did it, however only at 10 min. In Losina salt solution (physiological for Tetrahymena) the effect was weaker, however outer amino acid source was not absolutely needed for the production of the hormone. The results show that the TSH regulation of thyroid hormone synthesis (storage, secretion) and troph-hormone overlap can be deduced to a unicellular level. This may allow the hypothesis that the endocrine mechanisms proved at a low level of phylogeny are preserved for the higher ranked organisms.  相似文献   

10.
A single exposure of Tetrahymena to diiodotyrosine stimulated replication of the unicellular organism significantly relative to the control. The stimulatory effect tended to decrease with progressing time, but was still demonstrable after as many as 500 generations. Reexposures to diiodotyrosine also enhanced cell growth, indicating the existence of a receptor 'memory' in respect to the initial exposure, but their effect tended to decline soon after initial stimulation, and did not, in all probability, contribute to the preservation of the 'memory' which itself tends to vanish gradually in due course.  相似文献   

11.
Studies based on treatment with antibodies to thyrotropic hormone, luteotropic hormone, growth hormone or adrenocorticotropic hormone have shown that although the unicellular Tetrahymena does not possesssui generis receptors to all polypeptide hormones, such binding structures may arise, or become established in the membrane of the unicellular Tetrahymena in the presence of exogenous hormone. The Tetrahymena subjected to hormonal imprinting still contained an increased amount of hormone after six generation changes, which suggested that either hormone production had been induced by treatment, or the internalized hormone had not been degraded intracellularly. Thus the role of hormonal imprinting in receptor formation has also been substantiated by the immunocytochemical approach used in the present study.  相似文献   

12.
Diiodotyrosine induced receptor 'memory' at a concentration as low as 10(-18) M. Repeated exposure enhanced cellular responsiveness. Treatment with diiodotyrosine for 1 h, 4 times, induced receptor 'memory' more efficiently than a single uninterrupted treatment for 4 h. Immediately after induction, the receptor 'memory' is subject to retroactive interference by foreign hormonal stimuli, and can be extinguished completely by combined hormone treatment. Thus, the phenomenon of retroactive interference also takes effect at the unicellular level. The experimental observations indicate that receptor 'memory', induced in Tetrahymena by hormonal imprinting, has certain common features with the neuronal memory of higher organisms.  相似文献   

13.
Earlier experiments demonstrated that a membrane pattern of Protozoa behaves as a receptor with respect to hormones of higher organisms. This raised the possibility that some selection or strengthening of this unspecific pattern is involved in the evolution of the specific membrane patterns of the individual cells of higher organisms. In the present experiments, as a result of continual histamine treatment, the phagocytotic ability of a population of Tetrahymena was increased more intensely than after a single histamine treatment. The phagocytotic rate remained high for some time after the animals were returned from histamine-containing to normal medium. Thus, from the experimental data, it appears likely that as the hormone appears, selection becomes involved in the proliferation of cells possessing receptors. This observation might not only be of phylogenetic interest, but could also be relevant in receptor maturation as manifested in ontogenetic membrane differentiation.  相似文献   

14.
Earlier experiments demonstrated that a membrane pattern of Protozoa behaves as a receptor with respect to hormones of higher organisms. This raised the possibility that some selection or strengthening of this unspecific patter is involved in the evolution of the specific membrane patterns of the individual cells of higher organisms. In the present experiments, as a result of continual histamine treatment, the phagocytotic ability of a population of Tetrahymena was increased more intensely than after a single histamine treatment. The phagocytotic rate remained high for some time after the animals were returned from histamine-containing to normal medium. Thus, from the experimental data, it appears likely that as the hormone appears, selection becomes involved in the proliferation of cells possessing receptors. This observation might not only be of phylogenetic interest, but could also be relevant in receptor maturation as manifested in ontogenetic membrane differentiation.  相似文献   

15.
Cytosol estrogen receptor (ERc) and nuclear estrogen receptor (ERN) levels were investigated in rat uteri under different conditions of hormonal exposure. The amount of directly assayable receptor was closely related to the serum concentration of 17 beta-[2,4,6,7-3H] estradiol ( [3H]E2). A double injection technique was established to maintain serum levels of [3H]E2 which were sufficient to saturate receptor sites. Under these conditions, stable ERC and ERN levels are maintained throughout the study period. 30% of the total ER remains cytoplasmic in localization despite continuous hormonal exposure. Properties of ERC and ERN after 6 h of continuous hormonal exposure were investigated and found to be different from receptors found in these subcellular compartments 30 min after hormone injections. ERC from uteri 30 min after injection showed a faster sedimentation coefficient than ERC prepared 6 h after hormone treatment. ERC after 6 h of hormonal exposure showed a reduction of binding to calf thymus DNA adsorbed on cellulose in a cell-free system. ERC 30 min after [3H]E2 treatment had a biphasic dissociation pattern consistent with two different receptor populations, whereas uterine ERC obtained after 6 h of in vivo exposure to estradiol showed virtually no dissociation at 22 and 28 degrees C. In contrast to ERC, ERN 6 h after hormone injection sedimented faster than ERN obtained 30 min after treatment. KCl extractable ERN obtained either at 30 min or 6 h posthormone treatment showed biphasic dissociation kinetics at 22 and 28 degrees C, whereas KCl nonextractable ERN showed virtually no dissociation. Virtually all of the specifically bound ligand in cytosol and nuclear preparations was proven to be authentic E2. We conclude that total cellular receptor is quantitatively conserved during 6 h of continuous hormonal treatment. Nuclear receptor loss is not a requisite for receptor-mediated steroid function, although important time-dependent changes in receptor properties in both cytoplasmic and nuclear compartments do occur.  相似文献   

16.
After cyclodextrin-coated 10(-6) m steroid hormone treatment for 3 days (hormonal imprinting), Tetrahymena cells and their media were analysed by radioimmunoassay for the same hormone and for the presence of the other two. In the absence of hormone treatment, the cells contained no detectable levels of the three steroids. By 2 days in fresh medium following exposure of cells to a 72 h pretreatment of each specific hormone, correspondingly high quantities of hydrocortisone and oestradiol, but lesser quantities of testosterone, were found in both the media and the cells. One week after treatment only traces of hydrocortisone were found, exclusively within the cells themselves. Oestradiol was present in measurable quantities in both cells and media, whereas testosterone was only present in the medium. The presence of the other two hormones to the one used in the pretreatment were not usually present, except that when testosterone had been given, some oestradiol was also detected at 48 h, suggesting Tetrahymena has a functional cytochrome P(450)aromatase.  相似文献   

17.
Tetrahymena pyriformis GL cells pretreated (imprinted) and not pretreated with insulin showed dissimilar quantitative relations of FITC-insulin binding. Displacement of FITC-insulin by unlabelled insulin was considerably less in the control than in the imprinted series. The curve for saturation of the binding sites with FITC-insulin resembled a true saturation curve. The imprinted cells bound considerably more hormone in a shorter time than the control cells at identical levels of exposure. The dissociation of bound hormone from the imprinted cells increased over the control at 23 degrees C, and to a still greater degree at 4 degrees C. The effect of the pH of the medium on the dissociation of bound FITC-insulin also differed between the imprinted and not imprinted cells. Thus the proposed cytofluorimetric assay of binding kinetics demonstrated the actual conditions of receptor activity, and indicated that the induced insulin binding sites of Tetrahymena behaved similarly to 'classical' receptors.  相似文献   

18.
We previously revealed that Tetrahymena actin can copolymerize with rabbit skeletal muscle actin whereas it has a very divergent primary structure and some unusual properties. To investigate the effects of coexistence of this unusual Tetrahymena actin in mammalian cells, we here transfected Tetrahymena actin gene on an expression vector into COS-1 cells. From the results of immunofluorescence microscopy, it was found that Tetrahymena actin expressed in COS-1 cells copolymerized with intrinsic actin, and it was conspicuously localized to the tips of microfilament core bundles in microspikes. On the other hand, increase in cell number tended to cease temporarily about 24 hr after transfection with Tetrahymena actin gene, implying the inhibition of cytokinesis by Tetrahymena actin coexistence.  相似文献   

19.
The RIA technique detected prostaglandin (PGF2) and human placetal lactogen (hPL) in Tetrahymena cultures grown in bacto tryptone + yeast extract medium which, however, itself contained these hormones. About one to two per cent of the total hormone content of the medium was demonstrated intracellularly. Treatment with diiodotyrosine (T2), which is known to stimulate the growth of Tetrahymena, was followed by a decrease in the intracellular prostaglandin level. Triiodothyronine and thyroxine were not detected in Tetrahymena or in the medium, and did not appear in it on induction with TSH either. In the light of these observations it might well be doubted that prostaglandin was native in Tetrahymena: the use of synthetic media, and/or a reliable demonstration of the hormone content of the growth medium is recommended for evidence of hormone biosynthesis by unicellular organisms.  相似文献   

20.
Tetrahymena pyriformis reportedly possesses binding structures for the vertebrate hormone insulin that are amplified in cells having prior exposure to the hormone. Conventional and confocal microscopic studies were conducted to verify the validity of the reports and to localize the binding sites. Logarithmic cultures were exposed to insulin concentrations of 0, 3, 6, and 12 micrograms/ml for 1 h (receptor induced, RI). After an additional culture period the cells were fixed, exposed to porcine insulin (antigen), immunocytochemically processed, and examined for staining intensity by video image analysis. Observations indicate that T. pyriformis does bind insulin whether or not the cells have prior exposure to insulin. Staining intensity increased at the two highest RI concentrations over 0 microgram/ml (P less than 0.01) but the staining intensity at 0 microgram/ml was not different from that at 3 micrograms/ml. The results confirm that T. pyriformis does bind insulin and that prior exposure to insulin increases the binding capacity for insulin in what may be a concentration-dependent manner. Confocal microscopy of RI cells that had been labeled with either fluorescein isothiocyanate-insulin or the immunocytochemical technique outlined above revealed labeling of the cytoplasm that appeared to be vesicular. Both techniques produced very similar labeling patterns when optical sections through the cells were viewed. Conventional fluorescence revealed ciliary labeling that could be decreased by incubation with excess unlabeled insulin. Further studies with the exo- mutant of T. thermophila, SB 255, showed that mucocyst discharge and capsule formation are not involved in insulin binding.  相似文献   

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