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Query: UNIPROT:Q9UIJ5 (Rec)
 58,342 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cyclic AMP and cyclic GMP, considered to be cell proliferation regulators, have been reported to fluctuate in proliferating fibroblasts in vitro. The objectives of this experiment were to study the localization, distribution and staining patterns of these cyclic nucleotides in mechanically-stressed, proliferating periodontal fibroblasts in vivo. Cat canines were tipped by force applications for 0 to 48 hours and serial sagittal sections of fresh frozen, unfixed, undecalcified jaws were processed immuno-histochemically for the localization of cAMP and cGMP. Periodontal tension sites were studied microscopically. Fibroblastic staining for cAMP, which was localized mainly in the cell periphery, did not change appreciably as a result of tension. However, staining intensity increased one hour after the application of force, decreased after 6 hours and increased again at 24 hours. Staining for cGMP, initially covering the entire cell area, was concentrated over nuclei three hours after onset of tension, and diffused over cell periphery and cytoplasm thereafter. Intensity of staining for cGMP was maximal at 3 hours and low at 12 and 48 hours. These results demonstrate that stress-induced fibroblastic responses in vivo involve alterations in staining intensity for both cyclic nucleotides which may correspond with fluctuations of these regulators, reported to occur in vitro in various stages of the mitotic cycle. Moreover, this technique enables the identification of the involved cells in a stimulated, non-synchronized cell population.
Anat Rec 1978 Nov
PMID:Immuno-histochemical localization of cyclic nucleotides in the periodontium: mechanically-stressed cells in vivo. 8 16

Previous experiments indicate that bone cells respond to external stimuli with fluctuations of cyclic nucleotide levels. The objective of this experiment was to study the response of alveolar bone to the application of tensile forces through an examination of the osteoblastic staining pattern for cAMP and cGMP. Cat canines were tipped by 80-g force for 0 to 48 hours. Fresh frozen, unfixed, undecalcified jaws were sectioned sagittally and stained immuno-histochemically for cAMP and cGMP. In tension sites, osteoblastic staining intensity for cAMP decreased gradually from one to three hours, and then increased by 24 hours. Intense staining for cGMP, visible in osteoblasts of all treated cats, peaked after three hours of treatment and then again at 24 hours. Generally, groups of cGMP-stained osteoblasts were found adjacent to unstained osteoblasts. The observed fluctuations in the osteoblasts staining pattern for cAMP and cGMP indicates involvement of these substances in the early response of osteoblasts to mechanical stimuli in vivo.
Anat Rec 1978 Nov
PMID:Immuno-histochemical localization of cyclic nucleotides in mineralized tissues: mechanically-stressed osteoblasts in vivo. 8 17

We have assessed the regulatory influence of human recombinant TSH (rec-hTSH) on its homologous receptor (TSHR) using a well characterized human fetal thyroid monolayer cell culture technique. Under the culture conditions employed, fetal human thyroid cells showed basal expression of TSHR-specific mRNA transcripts, and the addition of rec-hTSH (1 U/L) induced up to an 8-fold increase in specific mRNA over a 48-h observation period. This induction was simulated by bromo-cAMP in a dose-dependent manner, indicating that the stimulatory effect of rec-hTSH was active at the postreceptor level. Furthermore, there was no detectable increase in the transcription rate of the TSHR gene after stimulation with rec-hTSH for 12-36 h, although a marked increase in thyroglobulin-specific mRNA was observed. Rec-hTSH also had no influence on the half-life of TSHR-specific mRNA, which remained at approximately 16 h in the presence or absence of rec-hTSH. These data indicate that rec-hTSH induced up-regulation in human thyroid cell TSHR-specific mRNA and that the mechanism of this regulation was likely to be secondary to a posttranscriptional nuclear event involving changes in the regulation of primary unspliced mRNA for the TSHR.
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PMID:The positive regulation of human thyrotropin (TSH) receptor messenger ribonucleic acid by recombinant human TSH is at the intranuclear level. 157 98

We have assessed the bioactivity of newly available recombinant human TSH (rec-hTSH) using human fetal thyroid cells, with the longer term aim of assessing its use for clinical applications. Rec-hTSH caused a consistent and dose-related increase in thyroid monolayer cell cAMP release and human thyroglobulin (hTg) secretion, confirming its bioactivity. Repetitive studies (n = 5) allowed us to derive an estimated biopotency for the rec-hTSH preparation examined of 5.6 IU/mg compared to 10 IU/mg for commercially available bovine TSH for human use. The rec-hTSH had a bioimmune ratio of 0.55, similar to that of purified pituitary hTSH standards, Furthermore, rec-hTSH induced thyroid epithelial cell growth, as evidenced by a decrease in thyroid cell doubling time from 54 +/- 2.1 to 31 +/- 1.7 h (P less than 0.005). Hence, rec-hTSH is a potent glycoprotein hormone preparation when measured in a homologous human thyroid cell culture system. Rec-hTSH could serve as a future definitive International Standard and has the potential for a useful diagnostic and therapeutic reagent.
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PMID:Recombinant human thyroid-stimulating hormone: initial bioactivity assessment using human fetal thyroid cells. 185 Nov 84

During perinatal development, when the size of the Sertoli cell population is determined, Leydig cells produce beta-endorphin, a peptide which may interact with Sertoli cells to modify their FSH-responsiveness, as suggested by our previous work. The goal of the present study was first, to test directly the possibility that beta-endorphin modifies the proliferative response of neonatal Sertoli cells to FSH, and second, to gain information on a mechanism(s) involved in any observed effect. We treated isolated 6-day-old Sertoli cells with FSH or vehicle in vitro and measured their incorporation of exogenous, radiolabeled thymidine with quantitative autoradiography. After 2 days in culture with FSH, we detected a 10-fold increase in the rate of Sertoli cell proliferation. The level of cell division in these FSH-treated cultures was identical to that in other cultures exposed to cAMP under similar conditions. In addition, inclusion of beta-endorphin 3 hr prior to FSH or cAMP decreased the effect of the hormone by 50% but left the cAMP response unchanged. Thus, beta-endorphin acts on isolated, neonatal Sertoli cells at a point prior to intracellular production of cAMP to suppress their response to FSH. When other cultures were treated with pertussis toxin, a blocker of intracellular GTP-binding proteins such as Gi, before sequential addition of endorphin and FSH, the effect of beta-endorphin on FSH-responsiveness was abolished. Moreover, when other cultures were exposed to pertussis toxin in the absence of endorphin, followed by FSH, their response to the hormone was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1990 Mar
PMID:Endorphin suppresses FSH-stimulated proliferation of isolated neonatal Sertoli cells by a pertussis toxin-sensitive mechanism. 213 7

The stage-dependent action of follicle-stimulating hormone (FSH) in the rat seminiferous epithelium was investigated in microdissected 1 mm tubule segments, where the precise stage of the cycle was identified by a rapid screening method of live cell squash preparations. For distinction of stages I and II and the substages of VII, new criteria were used. The step 16 spermatids with rapid assembly of outer dense fibers leading to marked increase of flagellar thickness were used for distinction of stages I and II. The form and density of the cytoplasmic lobes of step 19 spermatids was used for recognition of substages of VII. Highest basal production of cyclic AMP (cAMP, measured by radioimmunoassay) was found in stage II of the cycle and stages XIV-I-VI had higher values than did stages VII-XIII. A decline occurred during stage VII and an increase at stage XIV. When stimulated with FSH, highest cAMP secretion was found in stage IV of the cycle; again, stages XIV-I-VI had higher values than did other stages. A small but significant (P less than .01) stimulation was found at substage VIId. FSH-stimulated and basal cAMP productions of different stages were compared, highest values were found at stages IV and XIII, and lowest, at stages VIIa-c and IX of the cycle. Since the FSH-dependent cAMP production is confined to Sertoli cells, and the number of these cells is constant per unit length of seminiferous tubules, the Sertoli cells are obviously under a stage-specific paracrine control by the surrounding spermatogenic cells. Specific steps in cell differentiation, such as spermatogonial proliferation, final maturation of the spermatids (stages I-VII), onset of meiosis (substage VIId), and completion of meiotic divisions (stage XIV) may be involved in this interaction.
Anat Rec 1990 May
PMID:Modulation of basal and FSH-dependent cyclic AMP production in rat seminiferous tubules staged by an improved transillumination technique. 216 28

The possibility that ovarian steroids may participate in the inhibition of meiosis has not been rigorously examined. Since progesterone levels are extremely high in follicular fluid prior to ovulation, we tested the possibility that this steroid may be involved in oocyte maturation. To this end, we collected follicular oocytes and cultured them in the presence of dibutyrl cAMP (Bt2), progesterone, and/or the progesterone antagonist RU486 and assessed maturation evidenced by germinal vesicle breakdown (GVBD). Denuded oocytes or cumulus masses collected in the presence of 1 mM Bt2 and subsequently cultured in 25 microM progesterone did not undergo GVBD. However, denuded oocytes and cumulus masses collected in the presence of progesterone and not Bt2 did undergo GVBD (93%). Concentrations of Bt2 (150 microM) that would not inhibit GVBD were inhibitory when used in the presence of progesterone (1-25 microM). Competition experiments using increasing concentrations of the progesterone antagonist RU486 (1-100 microM) did not block the ability of progesterone to enhance the activity of Bt2. We conclude that progesterone alone does not block GVBD; however, in the presence of low concentrations of cAMP it is extremely effective in blocking GVBD. The synergistic activity of progesterone does not appear to be mediated by the progesterone receptor. The data suggest that progesterone and cAMP may operate cooperatively to inhibit meiosis in the ovarian follicle.
Anat Rec 1989 Apr
PMID:Effects of progesterone and a progesterone antagonist (RU486) on germinal vesicle breakdown in the mouse. 254 Jun 78

Some children with juvenile hypothyroidism exhibit unexplained precocious puberty. Interaction of TSH with the human FSH receptor (hFSH-R) is a possible pathophysiological mechanism for this syndrome that has not been explored due to the lack of hFSH-free TSH preparations and the scarcity of a suitable hFSH-R-based assay system. To devise an in vitro FSH bioassay suitable for exploring this mechanism, we expressed hFSH-R complementary DNA in COS-7 cells and stimulated them with recombinant hTSH (rec-hTSH). Rec-hTSH elicited a dose-dependent cAMP response in the in vitro hFSH-R bioassay; however, the concentration of rec-hTSH required for half-maximal stimulation was several logs greater than that of hFSH. Rec-hTSH acted as a competitive inhibitor of hFSH at the hFSH-R, indicating that hTSH and hFSH are acting through the same receptor, namely the hFSH-R. This provides a potential novel mechanism for the precocious puberty of juvenile hypothyroidism.
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PMID:A potential novel mechanism for precocious puberty in juvenile hypothyroidism. 762 56

Studies of human TSH (hTSH) structure and function have been limited by difficulties in producing large quantities of recombinant hormone. We describe a system for the stable expression of high levels of recombinant human TSH (rec hTSH) using a mutant form of dihydrofolate reductase (dhfr) as an amplifiable dominant selectable marker. A vector expressing both the hTSH alpha-subunit and the mutant dhfr was cotransfected with a hTSH beta-subunit expression vector into dhfr-deficient cells. Amplification of the transfected sequences by methotrexate selection, followed by cell culture in a hollow fiber perfusion system, yielded rec hTSH production as high as 100,000 microU/ mL. Immunoradiometric assays using five different antibodies revealed no differences in the immunological activities of rec hTSH and pituitary hTSH. Bioactivity was measured in a novel TSH bioassay coupling the generation of cAMP by a transfected hTSH receptor to the cAMP-dependent regulation of a luciferase reporter gene. The ED50 for bovine TSH in this bioassay was 1.4 ng/mL (3.5 x 10(-11) mol/L). The ratio of the ED50 values for rec hTSH and pituitary hTSH was 1.0:1.1 (P = NS), indicating that the two TSHs were of equivalent potency. In conclusion, we have developed techniques for the high level production of rec hTSH that is immunologically and biologically equivalent to pituitary hTSH. The ability to produce large quantities of rec hTSH using standard laboratory techniques should facilitate future studies, such as the development of clinically useful TSH analogs.
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PMID:Large scale synthesis of recombinant human thyrotropin using methotrexate amplification: chromatographic, immunological, and biological characterization. 877 98

This investigation examines the relationship between implantation strategy and gap junction protein expression in uterine endometrium. The pattern of gap junction and connexin protein expression was analyzed in porcine and equine endometrium from cycling and pregnant animals using electron microscopy and immunocytochemistry. Functional analysis of cell-cell communication was also monitored by laser cytometry in primary cultures of endometrial epithelial cells. Gap junctions were detected in endometrial stroma of cycling and pregnant animals, which was correlated with immunoreactive Cx43 within stromal fibroblasts and vascular elements. No Cx26, Cx32, or Cx43 immunostaining was detected in luminal endometrial epithelium in either the mare or the pig at any stage of the estrous cycle or pregnancy. In contrast, endometrial glands of the mare exhibited a spatiotemporal pattern of Cx43 expression in the apicolateral plasma membrane which, when present, colocalized with the tight junction-associated protein, ZO-1. Uterine glandular Cx43 expression in mares was present from day 3 postovulation through day 14 of diestrus and until day 23 of pregnancy, whereas Cx43 was absent within uterine glands during seasonal anestrus, estrus, and after day 30 of pregnancy. Primary cultures of equine endometrial epithelial cells expressed both immunoreactive Cx43 and significant gap junction-mediated intercellular communication (GJIC) which was rapidly upregulated by 1.0 mM 8-bromo-cAMP or blocked with 1.0 mM octanol. No GJIC or connexin protein was detected in cultured porcine epithelial cells despite incubation with a variety of agents, including 8-bromo-cAMP, steroid hormones, retinoic acid, and/or prolactin. Junctional communication in endometrial epithelium of domestic farm animals is different than that reported for species exhibiting invasive implantation. The absence of GJIC in uterine luminal epithelium of the gilt and mare may be involved in limiting trophoblast invasiveness.
Anat Rec 1998 07
PMID:Endometrial connexin expression in the mare and pig: evidence for the suppression of cell-cell communication in uterine luminal epithelium. 966 53


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