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PubMed2005

Expression of variant luteinizing hormone/chorionic gonadotropin receptors and degradation of chorionic gonadotropin in human chorionic villous macrophages.

Human gonads and non-gonadal organs/tissues express luteinizing hormone/chorionic gonadotropin (LH/CG) receptors. This study aimed to identify the LH/CG receptors and to clarify their function in human placental chorionic villous macrophages. Macrophages as well as syncytiotrophoblasts of human chorionic villous tissues were immunohistochemically positive for LH/CG receptor throughout gestation. By reverse transcription-nested polymerase chain reaction methods, villous macrophages were shown to express a variant type of LH/CG receptor, the sequencing of which revealed a deletion of exon 9. For experiments in vitro, a monocyte-macrophage cell line, THP-1, was transfected with vector alone, wild-type LH/CG receptor, and exon 9-deleted LH/CG receptor after phorbol 12-myristate 13-acetate (PMA) treatment. Non-PMA-treated THP-1 cells transfected with vector alone were also examined. THP-1 cells expressed exon 9-deleted LH/CG receptor after treatment with PMA. After the cells of the four groups were cultured in medium containing intact human CG (hCG), the concentrations of hCG and its beta-core fragment (beta-CF) were measured in the supernatant of the culture medium and in the cell cytosol. Time-dependent hCG uptake was observed in both non-PMA-treated and PMA-treated THP-1 cells, suggesting that the variant receptor is not directly involved in the ingestion of hCG. The degradation of hCG and excretion of beta-CF were progressed in PMA-treated cells but not in the un-treated cells. In the cell cytosol, the ratio of beta-CF and hCG concentrations (beta-CF/hCG) was significantly higher in the PMA-treated cells than in non-PMA-treated cells; however, it did not differ between the PMA-treated cells transfected with exon 9-deleted receptor and those transfected with vector alone. Macrophages may express the variant receptor in order to recognize the intracytoplasmic hCG and transport it to the lysosome. Among the two PMA-treated cells, the ratio was lower in those transfected with wild-type receptor. The expression of the variant receptor may modulate the degradation of hCG but be reduced by expression of the wild-type receptor in its lacking macrophages. Our data suggest a potentially important role for exon 9-deleted LH/CG receptors expressed in human placental villous macrophages in the local metabolism of hCG.

Base Sequence↗

PubMed1988

Double immunoelectron microscopic labelings of human chorionic gonadotropin and human placental lactogen in human chorionic villi.

Simultaneous immunoelectron microscopic localization of human chorionic gonadotropin (HCG) and human placental lactogen (HPL) was examined on the same ultrathin section of human chorionic villi by means of the double labeling technique. Using specific rabbit antisera against HCG and HPL followed by goat anti-rabbit IgG-coated colloidal gold of different sizes (15 nm and 5 nm), immunoreactions of HCG were concentrated on middle-sized secretory granules of 200-300 nm and large dense bodies of 500-1000 nm, while those of HPL were exclusively located on small secretory granules of 80-180 nm. The present experiment provides direct evidence for our previous data that HCG and HPL seem to be stored in different granular components in the syncytiotrophoblast.

Chorionic Gonadotropin↗

PubMed1988

Immunohistochemical localization of thrombomodulin in chorionic diseases of the uterus and choriocarcinoma of the stomach. A comparative study with the distribution of human chorionic gonadotropin.

Thrombomodulin (TM) is a newly described protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. TM is found on endothelium of the blood and lymph vessels and on syncytiotrophoblast of the placenta in humans. In the current study, we localized TM by immunohistochemistry in chorionic diseases of the uterus and choriocarcinomas of the stomach. The syncytiotrophoblast of hydatidiform mole contained TM. Uterine choriocarcinoma also had TM in the neoplastic syncytiotrophoblastic cells, although some of the neoplastic syncytiotrophoblastic cells showed no TM expression. Gastric choriocarcinoma never showed TM expression in the neoplastic syncytiotrophoblastic cells. However, all of the syncytiotrophoblastic cells had human chorionic gonadotropin (hCG) in hydatidiform mole, uterine choriocarcinoma, and gastric choriocarcinoma.

Biomarkers, Tumor↗

PubMed1989

Evolution of the autosomal chorion cluster in Drosophila. II. Chorion gene expression and sequence comparisons of the s16 and s19 genes in evolutionarily distant species.

We present a total of 13 kb of DNA sequences, encompassing autosomal chorion genes and their flanking DNA in four species of the genus Drosophila. Against a background of extensive divergence in introns and even in parts of the coding regions, islands of strong conservation are evident in the proximal 5' flanking and 5' untranslated sequences. An extragenic region of strong conservation is seen downstream of the last chorion gene in the autosomal cluster. The conserved DNA elements may be related to the conserved regulatory features of this cluster, including gene amplification and tissue- and temporally regulated transcription.

Amino Acid Sequence↗

PubMed1983

Isolation of laminin from human placental basement membranes: amnion, chorion and chorionic microvessels.

Laminin components were solubilized from basement membranes of amnion, chorion and chorionic microvessels of human placenta without prior protease digestion. These structures, after isolation, were initially processed in a sonicator bath containing a solution of Triton X-100, EDTA and 2M NaCl and the laminins extracted sequentially with 0.5M NaCl, 8 M urea, and 8 M urea + 2% 2-mercaptoethanol + 2% SDS. A high molecular weight (appr. 1 x 10(6)) complex containing laminins was purified by gel filtration on a Sepharose CL-2B column. This complex migrated as a single band on gel electrophoresis before reduction but resolved, after reduction, into four major laminin components, laminin A (350,000 M.W.), laminin M (240,000 M.W.) and laminins B1 and B2 (195,000 and 185,000 M.W.). Laminin M is a new molecular species of this protein.

Amino Acids↗

PubMed1984

Localization of a gene for a minor chorion protein in Drosophila melanogaster: a new chorion structural locus.

A minor chorion protein (called s70) with an approximate molecular weight of 70,000 D has been characterized in Drosophila melanogaster. The Staket geographic strain was found to carry an electrophoretic variant of this eggshell component and was used to determine the chromosomal location of the s70 gene. Our results establish a new locus for a chorion gene near yellow on the X chromosome and represent the first mapping of a quantitatively minor eggshell protein.

Animals↗

PubMed1984

Daily rhythm of serum human chorionic gonadotropin and human chorionic somatomammotropin in normal pregnancy.

We have quantified the human chorionic gonadotropin (HCG) and human chorionic somatomammotropin (HCS) serum levels in normal pregnant women at 1-h intervals during 24 h, through radioimmunoassay procedures. The HCG levels were higher during the first trimester, although no circadian variations were detected. Some variations exceeded by ten fold; these variations decreased with advanced periods of gestation. No diurnal fluctuations in serum HCS were apparent; in a case nearly at term, there was an increase from 05:00 h to 09:00 h simulating a circadian pattern, although it was not statistically significant. These data indicate that the placental protein secretion is autonomous and that HCG levels of fluctuation during early stages of pregnancy might be regulated by factors other than the trophoblast.

Chorionic Gonadotropin↗

PubMed1989

Gene evolution and regulation in the chorion complex of Bombyx mori. Hybridization and sequence analysis of multiple developmentally middle A/B chorion gene pairs.

Twenty-two pairs of chorion genes belonging to the A and B multigene families have been characterized and mapped within two segments of a 320 kb (1 kb = 10(3) bases or base-pairs) chromosomal walk in the domesticated silkmoth Bombyx mori. Eighteen of the gene pairs belong to two groups that are typified by the previously characterized A/B.L12 and A/B.L11 chorion gene pairs, and are defined by two respective types of short (approx. 280 base-pairs) bidirectional promoter sequences. In the chromosome, the L12-like and L11-like pairs are interspersed with each other and with the remaining four gene pairs, which have unrelated promoter sequences. We have sequenced the promoter regions and adjacent small exons of all L12-like and L11-like A and B genes in the walk. The L12-like promoters are highly conserved, whereas L11-like promoters are somewhat more variable. Reconsideration of previous data on RNA accumulation and disappearance during choriogenesis, in the light of the sequences, indicates that L12-like genes are developmentally early-middle, while L11-like genes correspond to two developmental subgroups, middle I and middle II. Sequence comparisons of all these promoters, as well as the previously characterized promoters of the developmentally late HcA and HcB genes, identify short elements of possible regulatory significance. The sequences, as well as extensive cross-hybridization analysis with short probes derived from the reference A/B.L12 gene pair, under carefully controlled conditions of stringency, indicate the occurrence of sequence transfers among A or B genes. These sequence transfers, which could result from gene conversions or unequal crossovers, are less abundant than in the HcA and HcB families, but do result in a patchwork of similarities and differences in the A and B genes. The transfers appear to be least frequent between the moderately divergent A genes that belong to different temporal classes, while the L12-like and L11-like B genes appear to be extensively homogenized in sequence.

Animals↗

PubMed2000

Adverse pregnancy outcome following post-chorionic villus sampling amniocentesis compared to chorionic villus sampling.

OBJECTIVE: To assess the adverse pregnancy outcome of post-chorionic villus sampling (CVS) amniocentesis and chorionic villus sampling. METHODS: Adverse pregnancy outcomes of 32 post-CVS amniocentesis cases and 264 CVS only cases were compared. The base-line characteristics were comparable in the 2 groups. RESULTS: One (3.1%) chromosomal abnormality was detected in the post-CVS amniocentesis group, compared to 5 (1.8%) in the CVS only group (p > 0.05). The fetal loss rate (spontaneous abortions and stillbirths) among continuing pregnancies was 3.2% in the post-CVS amniocentesis group and 3.5% in the CVS only group (p > 0.05). No statistically significant difference was found in the incidence of neonatal death, preterm delivery, fetal growth restriction, or congenital anomalies between the 2 groups. CONCLUSION: Adverse pregnancy outcome occurred at a similar frequency in the post-CVS amniocentesis group as in the CVS only group. Therefore, a subsequent amniocentesis after CVS can be considered as a safe procedure that does not introduce any additional adverse pregnancy outcome compared to that of CVS only.

Adult↗

PubMed1984

A comparative study of ectopic production of human chorionic gonadotropin and human chorionic somatotropin by carcinoma of the uterine cervix.

A comparative study was made on the ectopic production of chorionic gonadotropin (hCG) and chorionic somatotropin (hCS) by carcinoma of the uterine cervix. Incidence of hCG in the sera of cancer patients was much higher than hCS. hCS has a better prospect in early detection of cancer cervix than hCG. All hCS-producing tumors and the majority of hCG-producing ones were histologically diagnosed as squamous cell carcinoma. The source of hCG and hCS were found to be the tumor cells. A fall or disappearance of detectable hCG and hCS was noted following surgery or radiotherapy.

Adenocarcinoma↗

PubMed2005

Comparative characterisation of a commercial human chorionic gonadotrophin extracted from human urine with a commercial recombinant human chorionic gonadotrophin.

BACKGROUND: A commercial preparation of recombinant human chorionic gonadotrophin (r-hCG, Ovitrelle) was launched in 2001. Generally, hCG is available in two formats: human chorionic gonadotrophin (u-hCG), derived from the urine of pregnant females, and r-hCG produced by DNA based biotechnology. METHOD: The analytical characteristics of a highly purified u-hCG (Gonasi HP) were assessed and compared, for the first time, with the recombinant derived r-hCG (Ovitrelle). Gonasi HP is produced by extracting and purifying hCG from urine to obtain a specific bioactivity of 5000 IU/mg protein. Ovitrelle is produced via a recombinant derived mammalian cell line and purified to obtain a specific activity of 26 000 IU/mg. RESULTS AND CONCLUSION: It has been documented that commercially available u-hCG preparations can contain a number of urine derived protein contaminants as well as hCG related metabolites. This is also the case for Gonasi HP, where hCG related molecules and other proteins were found to be present, including epidermal growth factor (EGF) and eosinophil derived neurotoxin (EDN). It was also demonstrated that this preparation contained high levels of oxidised hCG. r-hCG was confirmed to be essentially intact hCG, free from contaminant proteins and with very low levels of oxidised hCG.

Chorionic Gonadotropin↗

PubMed1977

Chorionic gonadotropin, chorionic somatomammotropin, and prolactin in the uterine vein and peripheral plasma of pregnant rhesus monkeys.

Seven adult female rhesus monkeys were laparotomized at days 22, 42, and 157 of pregnancy and blood was collected from the uterine vein and peripheral circulation. Plasma samples were analyzed for monkey chorionic gonadotropin (mCG), monkey chorionic somatomammotropin (mCS), and prolactin by radioimmunoassay. Levels of mCG at day 22 of pregnancy were approximately 250 ng/ml; however, during the later stages of gestation mCG was either nondetectable or less than 0.7 ng/ml. There was no statistical difference in prolactin concentrations between days 22 and 42 of pregnancy, mean levels being between 176-424 ng/ml, but by day 157 prolactin levels of greater than 2,000 ng/ml were recorded. No statistical difference existed between peripheral and uterine vein concentrations of either mCG or prolactin at any of the stages of gestation examined. At day 22, mCS was not detectable; however, at day 42 of gestation mCS titers averaged 1.5 micrograms/ml and 2.3 micrograms/ml in the peripheral and uterine vein plasma, respectively. A statistically significant mCS increase occurred by day 157, levels in the periphery and uterine vein averaging 11.0 micrograms/ml and 16.3 micrograms/ml, respectively. Uterine vein titers of mCS were significantly higher than peripheral titers at both days 42 and 157. Thus, the highest levels of mCG were present during early pregnancy, whereas the highest levels of mCS and prolactin were present during late pregnancy.

Animals↗

PubMed1992

Regulation of chorionic gonadotropin-alpha and chorionic somatomammotropin messenger ribonucleic acid expression by 8-bromo-adenosine 3',5'-monophosphate and dexamethasone in cultured rhesus monkey syncytiotrophoblasts.

We wished to establish an in vitro culture system to examine gene expression in the context of differentiated function with rhesus monkey syncytiotrophoblasts. Chorionic villous tissue from placentas obtained at cesarean section was dispersed with trypsin and DNase and fractionated on a 5-70% Percoll gradient. When placed in culture, cells from a mononuclear fraction demonstrated to be very highly enriched (95-97% pure) for cytotrophoblasts aggregated and began to form syncytia within 24 h in culture, reminiscent of placental syncytiotrophoblast formation. The migration and fusion of individual cytotrophoblasts to form multinuclear syncytia were documented with time-lapse video microscopy. Incorporation studies with tritiated thymidine supported the conclusion from videomicroscopy that syncytia form by the fusion of individual cells and the addition of mononuclear cells to existing syncytia, rather than by endomitosis. The syncytiotrophoblast marker pregnancy-specific beta 1-glycoprotein (SP1) was immunocytochemically identified in both intact placenta and cultured syncytiotrophoblast cells. With cells isolated from placentas obtained on day 28, 50, 70, or 140 of pregnancy, treatment with 8-bromo-cAMP increased both rhesus monkey CG alpha-subunit (mCG alpha) and chorionic somatomammotropin (mCS) mRNA levels by an average of 4-fold. Increases of up to 2.5-fold were seen with mCG alpha mRNA in as little as 2 h after treatment, with a statistically significant average response seen within 6 h. The response with mCS required at least 24 h before a significant effect was seen. Actin mRNA levels were generally unchanged or suppressed by this treatment, indicating that the effect of 8-bromo-cAMP is relatively specific for the hormone mRNAs. Treatment of syncytiotrophoblasts with dexamethasone, but not progesterone or androstenedione, resulted in an approximately 4-fold increase in mCG alpha mRNA levels within 6 h of treatment. Steroid treatment did not affect mCS mRNA levels. Treatment with 4.5-400 nM GnRH or 0.1 to 100 ng/ml basic fibroblast growth factor likewise had no effect on the level of either mRNA, suggesting that any actions of these factors on hormone secretion are not effected via changes in steady state mRNA. These results communicate that the expression of the mRNAs for rhesus monkey CG alpha and CS in syncytiotrophoblast are regulated by steroid hormone- and cAMP-mediated pathways.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1990

Evolution of the autosomal chorion cluster in Drosophila. III. Comparison of the s18 gene in evolutionarily distant species and heterospecific control of chorion gene amplification.

We present a total of 6.1 x 10(3) base-pairs of DNA sequences, encompassing the s18 gene and flanking regions within the autosomal chorion cluster of three Drosophila species. Against a background of extensive divergence in the intron and even in parts of the coding region, islands of strong sequence conservation are evident. These are particularly notable in the 5' flanking DNA where they extend to approximately -600 base-pairs from the transcription start site. The most conserved segment of the entire chorion cluster is 71 base-pairs in the s18 5' flanking DNA, which in D. melanogaster is part of a region defined functionally as containing amplification control elements (ACE3 region). Transformation analysis, using chimeric transposons of D. melanogaster and D. grimshawi DNA, revealed that amplification control elements of D. grimshawi can support amplification in D. melanogaster. The functionally defined ACE3 region of D. grimshawi includes the conserved 71 base-pair segment, but also non-conserved sequences further upstream, which apparently enhance amplification.

Amino Acid Sequence↗

PubMed2001

The chorion genes of the medfly. II. DNA sequence evolution of the autosomal chorion genes s18, s15, s19 and s16 in Diptera.

We present a total of approximately 15 kb of DNA sequences, encompassing four chorion genes Ccs18, Ccs15, Ccs19, Cc16 and their flanking DNA in the medfly C. capitata. Comparison of coding regions, introns and intergenic sequences in five Dipteran species, D. melanogaster, D. subobscura, D. virilis, D. grimshawi and C. capitata documented an extensive divergence in introns and coding regions, but few well conserved elements in the proximal 5' flanking regions in all species. These elements are related to conserved regulatory features of three of the genes, including tissue- and temporal regulation. In the fourth, gene s15, significant alterations in the 5' flanking region may be responsible for its changed temporal regulation in C. capitata. One long intergenic sequence, located in the distal 5' flanking region of gene s18, is homologous to ACE3, a major amplification control element and contains an 80-bp A/T-rich sequence, known to stimulate strong binding of the origin recognition complex (ORC) in D. melanogaster. Analysis of the nucleotide composition of all chorion genes in C. capitata and D. melanogaster showed that C. capitata exhibit less biased representation of synonymous codons than does D. melanogaster.

Animals↗

PubMed2002

The Northwestern Twin Chorionicity Study: I. Discordant inflammatory findings that are related to chorionicity in presenting versus nonpresenting twins.

OBJECTIVE: The purpose of this study was to evaluate the association between chorionicity and discordant chorioamnionitis and funisitis in twin gestations. STUDY DESIGN: This was a retrospective analysis of 1156 twin placentas with a standardized diagnosis of chorionicity, chorioamnionitis, and funisitis for comparison between presenting and nonpresenting twins with dichorionic-separate, dichorionic-fused, and monochorionic placentas. RESULTS: Frequencies of chorioamnionitis in the nonpresenting twin were significantly lower in dichorionic placentas (odds ratio, 0.4; 95% CI, 0.3, 0.6, in dichorionic-separate placentas; odds ratio, 0.5; 95% CI, 0.3, 0.8, in dichorionic-fused placentas) compared with monochorionic placentas. The frequency of advanced inflammation (ie, chorioamnionitis with funisitis) was significantly lower in the nonpresenting twin than in the presenting twin, but only in dichorionic-separate placentas (odds ratio, 0.2; 95% CI, 0.1, 0.4). CONCLUSION: Dichorionic placentas confer significant protection against the spread of chorioamnionitis from the presenting to the nonpresenting gestational sac. In the more advanced process that involves the umbilical cord, only the subset of separate dichorionic placentas confers this protective effect against the spread of inflammation.

Adult↗

PubMed1988

Changes in circulating alphafetoprotein and human chorionic gonadotrophin following chorionic villus sampling.

Chorionic villus sampling (CVS) is rapidly becoming established as a routine procedure for first-trimester fetal diagnosis. The technique can result in fetomaternal haemorrhage and this might sensitize Rhesus-negative mothers and on occasion lead to spontaneous abortion. Serial sampling indicates that there is a rapid rise in alphafetoprotein (AFP) levels following CVS; however, this is not reflected by raised levels at 16-18 weeks and does not influence the subsequent pregnancy outcome. Unlike AFP, alterations in hCG levels are small and variable. Anti-D prophylaxis for non-sensitized Rhesus negative mothers should be given after CVS and the procedure may be contra-indicated in patients who are already sensitized.

Chorionic Gonadotropin↗

PubMed2005

Importance of the detection method for intact dimeric human chorionic gonadotropin without interference with the free human chorionic gonadotropin beta subunit for pregnancy exclusion before liver transplantation in a woman with cholangiocarcinoma.

Assay of human chorionic gonadotropin (hCG) is mainly used for the detection and monitoring of pregnancy, and for the follow-up of trophoblastic tumors. The serum free beta-hCG subunit (hCGbeta) is also a tumor marker in many non-trophoblastic tumors, including gastrointestinal cancers. In this work, we compared the performance of several immunoassays for pregnancy exclusion before liver transplantation and in the follow-up of a woman with cholangiocarcinoma. Serum hCG was detected with the Abbott Testpack plus hCG-Combo and measured with four automated sandwich immunoassays: ADVIA-Centaur, ACS:180, AxSYM and Dimension. hCGbeta was determined by an automated fluorescence sandwich immunoassay (Kryptor-Free beta hCG) and with a specific immunoradiometric assay (ELSA-F beta hCG, Schering). The expression of hCG was also evaluated by immunohistochemistry on sections of intrahepatic cholangiocarcinoma cells and on peritoneal metastases. Before transplantation, discordant results were observed for pregnancy exclusion. Qualitative Testpack and Dimension tests detected no hCG-like immunoreactivity, whereas the ADVIA-Centaur, ACS:180 and AxSYM tests revealed positive levels. The same discrepancy was obtained in follow-up of the patient after liver transplantation. hCGbeta assay and immunohistochemical staining revealed tumor cell secretion of hCGbeta. In conclusion, a specific serum immunoassay for intact dimeric hCG without cross-reaction with hCGbeta should be adopted as routine policy for pregnancy exclusion before liver transplantation.

Adult↗