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PubMedDate not supplied

Effects of luteinizing hormone, follicle-stimulating hormone, and epidermal growth factor on expression and kinase activity of cyclin-dependent kinase 5 in Leydig TM3 and Sertoli TM4 cell lines.

We examined the effects of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and epidermal growth factor (EGF) on the expression and kinase activity of cyclin-dependent kinase 5 (Cdk5) in Leydig TM3 and Sertoli TM4 cell lines. Hormonal regulation of the expression and activity of Cdk5 by using normal and hypophysectomized rat testes was also investigated to elucidate its role. Cdk5 levels and kinase activity were significantly elevated in TM3 cells that were grown in the presence of 7.5% serum, EGF, or LH and were associated with an increase in testosterone production compared with controls. These increases were accompanied by an increase in proliferation of TM3 cells after treatment with serum or EGF but not with LH suggest that Cdk5 may be involved in cellular differentiation that is induced with LH treatment. In contrast, the presence of neither serum, EGF, nor FSH had a significant effect on Cdk5 activity levels in the Sertoli TM4 cell line, and there was no correlation with proliferative activity or transferrin levels. A significant decrease in Cdk5 expression and activity were noted in rat testis after hypophysectomy compared with normal rat testis and is associated with a simultaneous decrease in testosterone and transferrin levels. Immunohistochemical analysis revealed that Cdk5 was strongly expressed in the nuclei and cytoplasm of Leydig cells, Sertoli cells, spermatogonia, and peritubular cells of normal adult rat testis. After hypophysectomy, the pattern of Cdk5 staining differed markedly from that in normal rat testis and a profound reduction in staining of Cdk5 was observed in each tubule. Our results suggest that LH and EGF influence and modulate Cdk5 expression and activity in Leydig TM3 cells and may, conceivably, be involved in signal transduction cascades that are initiated by hormones or growth factors. Cdk5 in Sertoli TM4 cells is likely to possess some constitutive functions that are not affected by the cells' proliferation state. Moreover, Cdk5 is probably involved in the constitutive and hormonally stimulated activities of the rat testis, in addition to its involvement in cell proliferation.

Animals↗
PubMed1998

[Indirect sensitized upconversion in Tm3+ and Yb3+ codoped non-crystal pentaphosphate].

We report the blue upconversion luminescence of Tm3+ and Yb3+ codoped non-crystal pentaphosphate pumped by -798nm laser diode. The samples were directly excited to 3F4 level of Tm3+ ion. Two energy transfer processes between Tm3+ and Yb3+ contribute mainly to the population of 1G4 level of Tm3+. Phonon plays an important role in the upconversion process.

English Abstract↗
PubMed2005

[Direct sensitization up-conversion mechanism from Yb3+ to Ho3+ and Tm3+ in fluoride].

The direct sensitization up-conversion processes from Yb3+ to Ho3+ and Tm3+ in fluoride were studied. The properties of the up-converted spectra of Yb3+ /Ho3+ and Ho3+ /Yb3+ /Tm3+ co-doped fluorides pumped by a 980 nm laser were compared. Their up-conversion mechanism has been analyzed and discussed. Two strong up-conversion luminescence bands have been observed, which were the transitions of 5 F4 5 S2 (Ho3+) -->18 (Ho3+) in the Yb3+ /Ho3+ co-doped fluoride system and 3 H4 (Tm3+) --> 3 H6 (Tm3+) in the Ho+ /Yb3+ /Tm3+ co-doped fluoride system. The addition of Tm3+ into Yb3+ /Ho3+ co-doped fluoride decreases the up-converted emission intensities of Ho3+ in the visible region.

Algorithms↗
PubMed1985

A novel leukocyte differentiation antigen: two monoclonal antibodies TM2 and TM3 define a 120-kd molecule present on neutrophils, monocytes, platelets, and activated lymphoblasts.

We produced two hybridomas by fusion of mouse myeloma cells with splenocytes from a mouse immunized with the THP-1 human monocytoid leukemia cell line. Two cloned hybridoma cell lines, designated as TM2 and TM3, were obtained. They secreted antibodies against a unique cell surface antigen expressed on all normal peripheral blood monocytes, neutrophilic granulocytes, platelets, and mitogen-induced lymphoblasts, some cells from patients with immature-type lymphoid leukemias. However, the antibodies reacted neither with large numbers of peripheral blood lymphocytes nor with red cells. Cross-blocking studies showed that these monoclonal antibodies recognized the same or a nearly positioned antigen epitope. Immunoprecipitation of THP-1 cell extract with TM2 or TM3 under reducing and nonreducing conditions yielded a specific band of mol wt equal to 120,000 daltons. This determinant appeared to be involved in granulocyte chemotaxis, since neutrophilic granulocytes exposed to TM2 or TM3 showed a significant decrease in chemotaxis toward endotoxin-activated serum. These two monoclonal antibodies did not affect O2- release or luminol-dependent chemiluminescence of neutrophils. Moreover, they did not alter platelet aggregation induced by thrombin. TM2 and TM3 will provide a new reagent in defining the linkage between lymphoid and myeloid differentiation and intermyeloid development.

Acute Disease↗
PubMed2004

Effects of chloride introduction on up-conversion luminescence in Tm3+ -doped tellurite glasses.

Blue up-conversion luminescence properties in Tm3+ -doped lead chloride tellurite glasses have been studied under 980 nm excitation. The intense blue and relatively weak red emissions centered at 476 and 649 nm corresponding to the transitions 1G4 --> 3H6 and 1G4 --> 3H4 of Tm3+, respectively, are simultaneously observed at room temperature. The effect of PbCl2 on up-conversion intensity is observed and discussed, and possible up-conversion mechanisms are evaluated. The intense blue up-conversion luminescence of Tm3+ -doped lead chloride tellurite glasses may be a potentially useful material for developing up-conversion optical devices.

Cations↗
PubMed2005

Intense blue up-conversion luminescence in Tm3+/Yb3+ codoped oxyfluoride glass-ceramics containing beta-PbF2 nanocrystals.

Up-conversion luminescence properties of a Tm3+/Yb3+ codoped oxyfluoride glass-ceramics under 980 nm excitation are investigated. Intense blue emission centered at 476 nm, corresponding to 1G4-->3H6 transitions of Tm3+ was simultaneously observed in the transparent oxyfluoride glass ceramics at room temperature. The intensity of the blue up-conversion luminescence in a 1 mol% YbF3-containing glass-ceramic was found to be about 40 times stronger than that in the precursor oxyfluoride glass. The reason for the intense Tm3+ up-conversion luminescence in the oxyfluoride glass-ceramics is discussed. The dependence of up-conversion intensities on excitation power and possible up-conversion mechanism are also evaluated.

Ceramics↗
PubMed2004

Tryptophan substitutions reveal the role of nicotinic acetylcholine receptor alpha-TM3 domain in channel gating: differences between Torpedo and muscle-type AChR.

A recent tryptophan scanning of the alpha-TM3 domain of the Torpedo californica AChR demonstrated that this domain can modulate ion-channel gating [Guzman, G., Santiago, J., Ricardo, A., Martí-Arbona, R., Rojas, L., Lasalde-Dominicci, J. (2003) Biochemistry 42, 12243-12250]. Here we extend the study of the alpha-TM3 domain to the muscle-type AChR by examining functional consequences of single tryptophan substitutions at five conserved positions (alphaM282, alphaF284, alphaV285, alphaA287, and alphaI290) homologous to the alpha-TM3 positions that were recently characterized in the Torpedo AChR. Similarly to the Torpedo AChR, mutations alphaM282W and alphaV285W, which are presumed to face the interior of the protein, did not exhibit functional channel activity. Nevertheless, significant expression levels of these mutants were observed at the oocyte surface. In contrast to the Torpedo AChR, in the muscle-type AChR, tryptophan substitution at positions F284, A287, and I290 produces a significant increase in normalized macroscopic response. Single-channel recordings at low ACh concentration revealed that the increase in AChR sensitivity for the F284W, A287W, and I290W is due to an increase in the mean open duration. These results suggest that tryptophan substitution directly affects channel gating, primarily the channel closing rate. Our results suggest that residues facing the interior of the protein (i.e., alphaM282 and alphaV285) may similarly affect channel gating in Torpedo and muscle-type AChR. However, equivalent mutations (i.e., F284W and I290W) presumably facing the lipid environment display a very different functional response between these two AChR species.

Amino Acid Sequence↗
PubMed2005

A spectroscopic analysis of blue and ultraviolet upconverted emissions from Gd3Ga5O12:Tm3+, Yb3+ nanocrystals.

The spectroscopic behavior of gadolinium gallium garnet (Gd3Ga5O12, GGG) nanocrystals codoped with 1% each of Tm3+ and Yb3+ prepared via a solution combustion synthesis procedure was investigated. Initial excitation of the codoped nanocrystals with 465.8 nm (into the 1G4 state) showed a dominant blue-green emission ascribed to the 1G4-3H6 transition as well as red and NIR emissions from the 1G4-3F4 and 1G4-3H5/3H4-3H6 transitions, respectively. Excitation at this wavelength (465.8 nm) showed the existence of a Tm3+ --> Yb3+ energy transfer process evidenced by the presence of the 2F5/2-2F7/2 Yb3+ emission in the NIR emission spectrum. The decay time constants proved that the transfer of energy occurred via the 3H4 state. Following excitation of the Yb3+ ion with 980 nm, intense upconverted emission was observed. Emissions in the UV (1D2-3H6), blue (1D2-3F4), blue-green (1G4-3H6), red (1G4-3F4), and NIR (1G4-3H5/3H4-3H6) were observed and were the direct result of subsequent transfers of energy from the Yb3+ ion to the Tm3+ ion. Power dependence studies showed a deviation from expected values for the number of photons involved in the upconversion thus indicating a saturation of the upconversion process. An energy transfer efficiency of 0.576 was determined experimentally.

Journal Article↗
PubMed2002

[High efficiency and low threshold upconversion from IR to red for Er3+ and Tm3+ co-doped fluoride-oxide glass-ceramic].

In this paper, high efficiency and low threshold upconversion from IR to red is reported, for Er3+ and Tm3+ co-doped fluoride-oxide glass-ceramic under 978 nm LD excitation. The component of sample in experiment is 65GeO2-25NaF-8.5BaF2-1Er2O3-0.5 Tm2O3, and the prepared method is obtained. The upconversion emission spectra under 978 nm LD excitation is measured at room temperature. Analyzing it, we find that introduction of Tm3+ into Er3+ doped system preferentially quenches the green upconversion fluorescence from 4S3/2 level of Er3+ duo to the efficient cross-relaxation of 4I13/2-->4I15/2 (Er): 3H6-->3H4 (Tm) which can significantly reduce the upconversion efficiency from 4I13/2 level to the emitting 4S3/2 level, and the Tm3+ behaves as a good sensitizer of the red upconversion from the 4F9/2 level of Er3+ which is mainly populated by the cross-relaxation of 3H4-->3H6 (Tm): 4I11/2-->4F9/2 (Er). However, at low Er3+ concentration (2 mol%), it is impossible for strong red upconversion. X-ray analysis is done, there are lots of nanocrystallites in MFG glass-ceramic. So we think, this red upconversion is attributed to Er3+ enriched fluoride microcrystallites, which makes the cross-relaxation of 3H4-->3H6 (Tm): 4I11/2-->4F9/2 (Er) more effective, therefore their active optical properties may be optimised. In the end, the relationship between LD working current and intensity of upconversion luminescence is discussed, the results confirm that both red and green upconversion processes are consisted by two photons.

Ceramics↗
PubMed2004

[Luminescence properties and up-conversion mechanism of Er3+ -Yb3+ -Tm3+ co-doped CdF2: PbF2-based glass].

Under 980 nm excitation, we obtained five intense up-conversion luminescence bands in Er3+ -Yb3+ -Tm3+ co-doped CdF2: PbF2-based glass as follows: infrared (800 nm), red (645 nm), green (545 nm and 525 nm), blue (480 nm ) and violet (407 nm) up-conversion luminescence. With the addition of Tm3+ , the intensity of 480 nm blue luminescence increased notably compared with that of co-doped Yb3+ -Er3+ sample, which attributed to Tm3+ special energy level structure; logI-logP plot of the luminescence intensity versus pump power shows that 480 nm luminescence emission is a two-photon excitation process induced by the cooperative up-conversion between two Yb3+ ions and the slope of logI-logP plot decreases gradually with increasing pump power and shows a tendency to bend down. The up-conversion mechanism is analyzed in detail under 980 nm semiconductor laser excitation; the dependence of 480 nm up-conversion luminescence intensity on the pump power under the steady states is discussed by using rate equation and the results agree well with the experiments.

English Abstract↗
PubMed2004

[Up-conversion luminescence in nanocrystalline zirconia co-doped with Yb3+ and Tm3+ ions].

Nanocrystalline zirconia co-doped with Yb3+ and Tm3+ ions was prepared by co-precipitation. Up-conversion luminescence of the samples doped with different Tm3+ concentrations was investigated under a 980 nm LD excitation. When x = 0.2 and 0.4, the luminescences of the two samples were mostly the same: strong blue up-conversion peaked at 474 nm and very weak red up-conversion were observed, corresponding to the transitions of 1G4 --> 3H6 and 1G4 --> 3F4 respectively. However, there was no luminescence of x = 1. Blue up-conversion luminescence of 0.2 mol% Tm3+-ions-doped nanocrystalline zirconia under different annealing temperatures was investigated. Blue up-conversion emission grew stronger with annealing temperature increasing. With pumping current increasing, blue up-conversion emission grew stronger too. By studying the sources of blue up-conversion, we concluded that it was three-photon process.

Ions↗
PubMed2006

[Construction and expression of ATP50 fluorescent protein in TM3 mouse Leydig cells].

OBJECTIVE: To study the expression and localization of ATP50 by construction of ATP50-pEYFP-N1 in primary cultured mouse Leydig cells. METHODS: Primary cultured mouse Leydig cells were confirmed by 3B-HSD staining. ATP50 was cloned into pEYFP-N1 between Bam HI and Eco RI sites. Cell-transfection and living-cell fluorescence imaging microscopy were employed to investigate the sub-cellular localization of YFP-ATP50 in TM3 mouse Leydig cells. RESULTS: ATP50 green fluorescent protein was well co-localized with red fluorescence mitochondrion marker-Mitotracker in TM3 mouse Leydig cells. CONCLUSION: ATP50 was expressed in primary cultured mouse Leydig cells. The fluorescent expression vector of ATP50 was constructed successfully and YFP-ATP50 was located in mitochondria in TM3 mouse Leydig cells, which provided a useful clue for further research on the steroidogenesis dysfunction in aging males.

Adenosine Triphosphatases↗
PubMed2005

Growth suppression of Leydig TM3 cells mediated by aryl hydrocarbon receptor.

Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin induces developmental toxicity in reproductive organs. To elucidate the function of AhR, we generated stable transformants of TM3 cells overexpressing wild-type aryl hydrocarbon receptor (AhR) or its mutants which carried mutations in nuclear localization signal or nuclear export signal. In the presence of 3-methylcholanthrene (MC), proliferation of the cells transfected with wild-type AhR was completely suppressed, whereas cells expressing AhR mutants proliferated in a manner equivalent to control TM3 cells, suggesting AhR-dependent growth inhibition. The suppression was associated with up-regulation of cyclin-dependent kinase inhibitor p21Cip1, which was abolished by pretreatment with actinomycin D. A p38 MAPK specific inhibitor, SB203580, blocked the increase of p21Cip1 mRNA in response to MC. Treatment with indigo, another AhR ligand, failed to increase of p21Cip1 mRNA, although up-regulation of mRNA for CYP1A1 was observed. These data suggest AhR in Leydig cells mediates growth inhibition by inducing p21Cip1.

Animals↗
PubMed2005

Nicotine induces apoptosis in TM3 mouse Leydig cells.

OBJECTIVE: To investigate whether nicotine-induced testicular toxicity involves the induction of apoptosis in Leydig cells. DESIGN: Cell study using cells of the TM3 cell line derived from mouse Leydig cells. SETTING: Academic research laboratory. INTERVENTION(S): Morphological and biochemical analyses for the detection of apoptosis. MAIN OUTCOME MEASURE(S): The effect of nicotine on the occurrence of apoptosis was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6-diamidino-2-phenylindole staining, terminal deoxynuclotidyl transferase-mediated dUTP nick end labeling assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction, caspase-3 enzyme assay, and Western blot analysis. Nicotine treatment exhibits several features of apoptosis in mouse Leydig cells. RESULT(S): TM3 cells treated with nicotine exhibit several features of apoptosis. It was also shown that nicotine increases the mRNA level of bax and decreases that of bcl-2. In addition, nicotine enhanced the expression of the activated form of caspase-3 and caspase-3 enzyme activity. CONCLUSION(S): Nicotine appears to activate specific intracellular death-related pathways, probably by bax-dependent activation of caspase-3, inducing apoptosis in Leydig cells. Thus, nicotine-induced apoptosis of Leydig cells might be one of the important mechanisms behind nicotine-related urogenital disorders in men.

Animals↗
PubMed2005

Structural and up-conversion luminescence properties in Tm3+/Yb3+-codoped heavy metal oxide-halide glasses.

Tm3+/Yb3+-codoped heavy metal oxide-halide glasses have been synthesized by conventional melting and quenching method. Structural properties were obtained based on the Raman spectra, indicating that halide ion has an important influence on the phonon density and maximum phonon energy of host glasses. Intense blue and weak red emissions centered at 477 and 650 nm, corresponding to the transitions 1G4-->3H6 and 1G4-->3H4, respectively, were observed at room temperature. The possible up-conversion mechanisms are discussed and estimated. With increasing halide content, the up-conversion luminescence intensity and blue luminescence lifetimes of Tm3+ ion increase notably. Our results show that with the substitution of halide ion for oxygen ion, the decrease of phonon density and maximum phonon energy of host glasses both contribute to the enhanced up-conversion emissions.

Fluorescence↗
PubMed2005

Intense upconversion luminescence and effect of local environment for Tm3+/Yb3+ co-doped novel TeO2-BiCl3 glass system.

We present the results of a study that uses theoretical and experimental methods to investigate the characteristics of the upconversion luminescence of Tm3+/Yb3+ codoped TeO2-BiCl3 glass system as a function of the BiCl3 fraction. These glasses are potentially important in the design of upconversion fiber lasers. Effect of local environment around Tm3+ on upconversion fluorescence intensity was analyzed by theoretical calculations. The structure and spectroscopic properties were investigated in the experiments by measuring the Raman spectra, IR transmission spectra, and absorption and fluorescence intensities at room temperature. The results indicate that blue luminescence quantum efficiency increases with increasing BiCl3 content from 10 to 60 mol%, which were interpreted by the increase of asymmetry of glass structure, decrease of phonon energy and removing of OH- groups.

Bismuth↗
PubMed2005

Estrogenic compounds inhibit gap junctional intercellular communication in mouse Leydig TM3 cells.

Some estrogenic compounds are reported to cause testicular disorders in humans and/or experimental animals by direct action on Leydig cells. In carcinogenesis and normal development, gap junctional intercellular communication (GJIC) plays an essential role in maintaining homeostasis. In this study, we examine the effects of diethylstilbestrol (DES, a synthetic estrogen), 17beta-estradiol (E(2), a natural estrogen), and genistein (GEN, a phytoestrogen) on GJIC between mouse Leydig TM3 cells using Lucifer yellow microinjection. The three compounds tested produced GJIC inhibition in the TM3 cells after 24 h. Gradually, 10 microM DES began to inhibit GJIC for 24 h and this effect was observed until 72 h. On the other hand, both 20 microM E(2) and 25 microM GEN rapidly inhibited GJIC in 6 h and 2 h, respectively. The effects continued until 24 h, but weakened by 72 h. Furthermore, a combined effect at microM level between DES and E(2) on GJIC inhibition was observed, but not between GEN and E(2). DES and E(2) showed GJIC inhibition at low dose levels (nearly physiological estrogen levels) after 72 h, but GEN did not. DES-induced GJIC inhibition at 10 pM and 10 microM was completely counteracted by ICI 182,780 (ICl), an estrogen receptor antagonist. On the other hand, the inhibitory effects on GJIC with E(2) (10 pM and 20 microM) and GEN (25 microM) were partially blocked by ICI or calphostin C, a protein kinase C (PKC) inhibitor, and were completely blocked by the combination of ICI and calphostin C. These results demonstrate that DES inhibits GJIC between Leydig cells via the estrogen receptor (ER), and that E(2) and GEN inhibit GJIC via ER and PKC. These estrogenic compounds may have different individual non-genotoxic mechanism including PKC pathway on testicular carcinogenesis or development.

Animals↗
PubMed2002

Alcohol induces apoptosis in TM3 mouse Leydig cells via bax-dependent caspase-3 activation.

To investigate whether ethanol induces apoptosis in Leydig cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, terminal deoxynuclotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, DNA fragmentation assay, caspase-3 enzyme assay, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis were performed on TM3 mouse Leydig cells. Through morphological and biochemical analyses, it was demonstrated that TM3 cells treated with ethanol at concentrations of 50 and 100 mM exhibit classical apoptotic features. In addition, it was shown that ethanol induces increases in levels of bax and caspase-3 and a decrease in bcl-2 expression. Based on the results, alcohol appears to activate specific intracellular death-related pathways leading to bax-dependant caspase-3 activation and the induction of apoptosis in Leydig cells.

Animals↗