RESUMEN
Gynandroblastoma, an extremely rare ovarian tumour that usually consists of both Sertoli stromal cell and granulosa cell tumours, often produces both androgenic and estrogenic effects. The authors herein report a case of gynandroblastoma with the longest disease-free period reported to date. A 66-year-old woman without metrorrhagia or hirsutism presented with abdominal pain and slightly elevated serum estradiol levels. Her uterus was enlarged, and endometrial curettage performed to reduce endometrial thickness prior to laparotomy led to a diagnosis of atypical endometrial hyperplasia. She was diagnosed of ovarian tumour. The pathology report revealed that the right ovarian tumour was a "gynandroblastoma". Such lesions are classified as borderline malignant. Postoperative adjuvant therapy was not administered in this case because only a few recurrent or fatal cases have been reported. The lesion was classified as pTlaN0M0 according to Union for International Cancer Control (UICC). The patient is alive and has been disease-free for 77 months post-surgery.
Asunto(s)
Posmenopausia , Tumores de los Cordones Sexuales y Estroma de las Gónadas/patología , Anciano , Resultado Fatal , Femenino , Humanos , Estadificación de Neoplasias , Neoplasias Ováricas/diagnóstico por imagen , Neoplasias Ováricas/patología , Tumores de los Cordones Sexuales y Estroma de las Gónadas/cirugíaRESUMEN
The effects of intrauterine growth retardation (IUGR) on brain histological or functional development were examined in rats. IUGR was induced by ligating the bilateral uterine arteries at day 17 of pregnancy. On day 22 of pregnancy, cesarean section was performed, and pups with a birth weight of <2 SD of the mean birth weight of control pups were regarded as IUGR rats. Morphological changes of the brain were studied by Nissl's staining at different timepoints during prenatal and postnatal periods. For behavioral study, an open-field test was performed at 5, 7 and 10 weeks after birth. Histological studies showed the migration disorder of the neurons in the cerebral cortex from embryonic day 17 to postnatal day (PD) 49. The open-field test revealed locomotor disturbance at PD49 in male IUGR rats, but not in female IUGR rats or control rats. It is concluded that IUGR due to antenatal ischemia-hypoxia causes morphological changes in the central nervous system, and induces behavioral impairment, particularly in male rats.
Asunto(s)
Conducta Animal , Encéfalo/embriología , Encéfalo/crecimiento & desarrollo , Retardo del Crecimiento Fetal/complicaciones , Hipoxia-Isquemia Encefálica/complicaciones , Animales , Arterias/cirugía , Femenino , Ligadura , Masculino , Actividad Motora , Embarazo , Ratas , Ratas Sprague-Dawley , Caracteres Sexuales , Útero/irrigación sanguíneaRESUMEN
The human has two relaxins, termed H1 and H2, both of which are biologically active and co-expressed in the decidua, placenta and prostate; in the corpus luteum, the main source of circulating relaxin, only the H2 form is expressed. The reasons for this differential expression of the relaxin genes are unknown. The possibility that their 3'-untranslated regions (UTRs) contribute to this differential expression by affecting their mRNA stabilities was investigated. Thus the 3'-UTRs of both relaxin genes were isolated through a combined 3'-rapid amplification of cDNA ends-PCR (RACE-PCR) using poly (A)(+)RNA from human decidua, placenta, prostate and corpus luteum. The sequences obtained for each 3'-UTR were identical in the tissues examined, were AT-rich (72%) and showed 91% homology between relaxin H1 and H2 when maximally aligned to include several gaps, the significance of which is unknown. Relaxin H1 has two, and relaxin H2 has one, poly (A)(+) signal, in addition to one cytoplasmic polyadenylation element 30 nucleotides upstream of this. The mRNA levels of relaxin H1 and H2 in the prostate adenocarcinoma LNCaP.FGC cell line were determined by quantitative competitive RT-PCR. Relaxin H1 had a 10-fold greater number of molecules (approximately 2.5x10(7)) per microgram of total RNA than relaxin H2 (approximately 2.5x10(6)). The stability of relaxin H1 and H2 mRNAs were compared in LNCaP cells treated with the transcription inhibitor actinomycin D (10 mM) for 0, 1, 2, 4, 8, 10, 14, or 24 h. Half-lives of 3.17 days for relaxin H1 mRNA and 11. 4 h for relaxin H2 mRNA were obtained from semi-logarithmic plots. Thus both mRNAs are relatively stable; however, relaxin H1 mRNA is considerably more stable than relaxin H2, at least in LNCaP cells. This difference in their mRNA stability may partly explain the greater level of expression of relaxin H1 in these cells.
Asunto(s)
Regiones no Traducidas 3'/genética , Relaxina/genética , Secuencia de Bases , Cuerpo Lúteo/metabolismo , Dactinomicina/farmacología , Decidua/metabolismo , Membranas Extraembrionarias/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Masculino , Datos de Secuencia Molecular , Placenta/metabolismo , Reacción en Cadena de la Polimerasa/métodos , Embarazo , Próstata/metabolismo , Precursores de Proteínas/genética , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Transcripción Genética/efectos de los fármacosRESUMEN
OBJECTIVE: This study was undertaken to determine which genes were up-regulated by acute distention in an amniotic epithelial cell line and in human fetal membranes. STUDY DESIGN: WISH cells, a human amniotic epithelial cell line, were grown on silicone elastomer sheets coated with extracellular matrix and reproducibly distended by 40% in a novel device for 4 hours. Differential gene expression was analyzed by means of suppression subtractive hybridization. Expression of the identified genes was then quantitated by Northern blot analysis in fetal membrane explants after distention in the same device for 4 hours. The effect of distention on apoptosis of the cells and tissue samples was concomitantly studied by means of the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method. RESULTS: The genes for interleukin 8 and pre-B-cell colony-enhancing factor were found to be up-regulated in both the WISH cells and the distended fetal membranes. The apoptotic index values in both the cells and the tissue samples were unaffected by distention. CONCLUSIONS: Acute distention induces the up-regulation of interleukin 8 and pre-B-cell colony-enhancing factor in both WISH cells and human fetal membranes and does not cause apoptosis.
Asunto(s)
Amnios/citología , Amnios/fisiología , Regulación de la Expresión Génica , Apoptosis , Fenómenos Biomecánicos , Northern Blotting , Línea Celular , Medios de Cultivo , Citocinas/genética , Células Epiteliales/fisiología , Matriz Extracelular , Femenino , Humanos , Etiquetado Corte-Fin in Situ , Interleucina-8/genética , Nicotinamida Fosforribosiltransferasa , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Embarazo , ARN Mensajero/análisisRESUMEN
BACKGROUND: The search for the cause of chronic hepatitis among individuals with non-A to G hepatitis has led to the discovery of a post-transfusion hepatitis-related DNA virus, designated TT virus (TTV), which, based on viral sequences, belongs to a new virus family. The principal modes of infection with TTV are poorly understood, and its role in human immunodeficiency virus type 1 (HIV-1) infection is unclear. OBJECTIVE: To determine if injection drug use (IDU) and high-risk heterosexual activity (HRHA), principal modes of acquiring HIV-1 infection, place individuals at greater risk of acquiring TTV. METHODS: The authors analyzed DNA, extracted from sera or filter paper-blotted whole blood, obtained during August 1997 and June 1998 from 324 Vietnamese (148 male; 176 female), for TTV sequences by hot-start, heminested polymerase chain reaction. RESULTS: Prevalence of TTV viremia was similar among individuals engaging in IDU or HRHA (23.4% vs. 20.2%; P > 0.5), with no age- or gender-specific differences. No association was found between TTV viremia and co-infection with HIV-1 or hepatitis C virus (HCV). Phylogenetic analysis of 30 TTV sequences revealed two distinct genotypes and four subtypes that did not segregate according to gender, HIV-1 and HCV risk behaviors, or geographic residence. CONCLUSIONS: Among HIV-1- or HCV-infected Vietnamese, who presumably acquired their infection by either the parenteral or nonparenteral route, the data indicate no clear association between acquisition of TTV infection and risk behavior for HIV-1 or HCV infection, suggesting that the usual route of TTV transmission in Vietnam is other than parenteral or sexual.
Asunto(s)
Infecciones por Virus ADN/transmisión , Infecciones por VIH/complicaciones , Hepatitis C/complicaciones , Hepatitis C/transmisión , Hepatitis Viral Humana/transmisión , Adolescente , Adulto , Infecciones por Virus ADN/complicaciones , Infecciones por Virus ADN/epidemiología , Infecciones por Virus ADN/virología , Virus ADN/genética , Virus ADN/aislamiento & purificación , ADN Viral/análisis , Femenino , Infecciones por VIH/transmisión , Infecciones por VIH/virología , VIH-1 , Hepacivirus/genética , Hepacivirus/inmunología , Hepacivirus/aislamiento & purificación , Hepatitis C/virología , Virus de Hepatitis/genética , Virus de Hepatitis/aislamiento & purificación , Hepatitis Viral Humana/epidemiología , Hepatitis Viral Humana/virología , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Prevalencia , Asunción de Riesgos , Análisis de Secuencia de ADN , Abuso de Sustancias por Vía Intravenosa , Vietnam/epidemiología , Viremia/virologíaRESUMEN
OBJECTIVE: To determine whether suppression subtractive hybridization can detect genes in fetal membranes that are upregulated by infection, preterm premature rupture of membranes (PROM), or labor. METHODS: Using suppression subtractive hybridization, messenger RNAs from a preterm fetal membrane obtained at cesarean delivery without labor (control) were subtracted from a pool of messenger RNAs of three patients with preterm PROM and vaginal delivery. Eight candidate genes identified as upregulated were quantitated by Northern analysis in each of the tissues and in additional patient subgroups. RESULTS: Eight differentially upregulated genes were identified in preterm labor with PROM. Four of the genes are known to be involved in the response to inflammation or infection, and subsequent histologic examination showed one of the preterm PROM tissues to be infected. F-actin capping protein and chitinase precursor, not previously known to be involved in infection, were also upregulated in the infected tissue from preterm PROM. Northern blots using additional subgroups of patients showed that a regulatory G-protein signaling protein gene was significantly upregulated at term by labor in addition to significant upregulation of interleukin-8. There was a strong correlation between the gene expression for complement factor-B and duration of membrane rupture in the patients with preterm PROM. CONCLUSION: Two novel genes potentially involved in the response to inflammation or infection have been identified. A regulatory G-protein signaling protein and interleukin-8 gene expression were upregulated by labor. Complement factor-B gene expression was directly related to the duration of membrane rupture.
Asunto(s)
Membranas Extraembrionarias/fisiología , Rotura Prematura de Membranas Fetales/genética , Trabajo de Parto/genética , Trabajo de Parto Prematuro/genética , Complicaciones Infecciosas del Embarazo , Regulación hacia Arriba/genética , Adulto , Femenino , Humanos , Hibridación de Ácido Nucleico/métodos , EmbarazoRESUMEN
Suppression Subtractive Hybridization (SSH) has been used to compare rat Nb2 cells treated with prolactin for 1 hour with untreated cells. This new method for identifying differentially expressed genes showed that the mRNAs for at least three genes were elevated by such treatment, including a p38 mitogen activated protein (MAP) kinase. The p38 MAP kinase was cloned and the full length cDNA sequence was determined.
Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Linfoma de Células T/enzimología , Proteínas Quinasas Activadas por Mitógenos , Prolactina/farmacología , Animales , Northern Blotting , Clonación Molecular , ADN Complementario , Linfoma de Células T/patología , Hibridación de Ácido Nucleico , Ratas , Técnica de Sustracción , Células Tumorales Cultivadas , Regulación hacia Arriba , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
A Northern blot containing poly(A)+ RNA isolated from pregnant rabbit placenta, uterus, ovary, mammary gland, psoas muscle, and intestine was hybridized with an oligonucleotide probe to porcine preprorelaxin. Only the placenta and uterus exhibited high levels of a 1-kb mRNA that hybridized to the probe. Porcine preprorelaxin primers were used to generate and amplify cDNAs from placental and uterine poly(A)+ RNA. Southern blot analysis of the placental and uterine cDNAs each revealed a single PCR product that hybridized with the porcine relaxin probe. The sequence of the cDNAs exhibited 100% identity with SQ10, a preprorelaxin-like gene identified in rabbit tracheobronchial epithelial cells. Identical results were obtained with primers specific for SQ10. SQ10 is expressed in tracheobronchial epithelial cells during abnormal conditions, such as mechanical or toxic injury. The protein is believed to have a role in tracheal epithelial cell differentiation and transformation that occurs under the above conditions. This work shows, for the first time, the presence of high levels of message for SQ10 in normal reproductive tissues. The expression of SQ10 in the uterus and placenta may suggest an important role in cell differentiation and transformation that occurs in these tissues during pregnancy.
Asunto(s)
Expresión Génica , Placenta/metabolismo , Precursores de Proteínas/genética , Relaxina/genética , Útero/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Southern Blotting , ADN Complementario/análisis , ADN Complementario/química , Femenino , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Embarazo , Precursores de Proteínas/química , ARN Mensajero/análisis , ARN Mensajero/metabolismo , ADN Polimerasa Dirigida por ARN , Conejos , Relaxina/química , PorcinosRESUMEN
A novel member of the insulin superfamily has previously been shown to be expressed only in porcine pre and postnatal Leydig cells and its human analogue demonstrated in the human testes but not in other organs and hence has been tentatively termed Leydig insulin-like peptide (Ley I-L). However, we have detected hLey I-L gene expression in the cyclic human corpus luteum and trophoblast by the reverse transcriptase-polymerase chain reaction (RT-PCR), with primers selected from the published human Ley I-L sequence. Normal and neoplastic breast tissue and fetal membranes with adhering decidua did not express the gene. The overall sequence of the trophoblast gene was in agreement with that reported with minor changes only in the putative connecting peptide, confirmed by restricted enzyme digestion. A 290 bp RT-PCR product was cloned and used as a cDNA probe in Northern analyses; hybridization was readily shown with cyclic corpora lutea but not with other tissues. The broader spectrum of the expression of this gene will warrant a new nomenclature when its biological activities are known. The different intensity of expression in the corpus luteum and trophoblast suggest endocrine and autocrine/paracrine roles respectively in these tissues in which H2 relaxin and H1/H2 relaxins coexist respectively and at similar levels of expression. Operationally the amino acid sequence homologies between the processed H1 and H2 relaxins and hLey I-L may qualify the specificity claimed for immunostaining the human relaxins in the corpus luteum and trophoblast.
Asunto(s)
Cuerpo Lúteo/fisiología , Expresión Génica , Insulina/genética , Células Intersticiales del Testículo/fisiología , Familia de Multigenes , Trofoblastos/fisiología , Secuencia de Bases , Femenino , Humanos , Masculino , Sondas Moleculares/genética , Datos de Secuencia Molecular , Reacción en Cadena de la PolimerasaRESUMEN
Immunoreactive relaxin is present in human breast cyst fluid and postpartum milk without concurrent detectable serum levels, suggesting that the breast is a site of relaxin synthesis. Monoclonal and polyclonal antibodies to human relaxin H2 have been used to immunolocalize relaxins in normal, benign and neoplastic breast tissues with the avidin-biotin immunostaining technique. In view of the similarities in amino acid sequence between H1 and H2 relaxins, these antibodies to H2 relaxin are likely to detect either or both relaxins present in tissue sections. Staining patterns with these antibodies were identical and showed positive diffuse cytoplasmic staining in normal, lobular and ductal epithelium and in myoepithelial cells in breast tissues from normal prepubertal, cyclic, gestational, lactational and postmenopausal females. Relaxin staining was also present in epithelial and myoepithelial cells of ducts and lobules in benign breast disease as well as in metaplastic epithelium of apocrine microcysts. All breast carcinomas (infiltrating ductal, tubular, medullary, intraductal and infiltrating lobular carcinomas) had strong uniform cytoplasmic staining within the neoplastic epithelial cells. All staining was abolished in normal and neoplastic tissues when the polyclonal antibody was preabsorbed with relaxin. It was necessary to distinguish between the possibilities of relaxins being sequestered by breast tissue and local synthesis. Therefore, the expression of the H1, H2 or both human relaxin genes in normal and neoplastic breast tissues was studied by the isolation of RNA, synthesis of first strand cDNA and amplification by PCR using primer sets which amplified either both H1 and H2, or specifically only H1 or H2 relaxin. The coamplification of both relaxin genes was verified by Southern analysis, diagnostic restriction enzyme digestion and sequencing. The primer set for H1 relaxin detected H1 gene expression in 1 out of 8 normal and 9 out of 12 neoplastic breast RNA samples. The H2 relaxin gene was found to be expressed in 3 out of 8 of the normal samples but in all 12 of the neoplastic samples, suggesting that this gene is expressed at higher copy number in the neoplastic tissues. This is the first demonstration of the cellular immunolocalization of relaxin and relaxin gene expression in normal and neoplastic breast. This should allow further exploration of relaxin's role(s) in normal breast physiology and in its tumorigenesis.
Asunto(s)
Neoplasias de la Mama/metabolismo , Mama/metabolismo , Carcinoma/metabolismo , Relaxina/metabolismo , Adulto , Factores de Edad , Anticuerpos Monoclonales/inmunología , Secuencia de Bases , Northern Blotting , ADN Complementario/genética , Femenino , Humanos , Técnicas para Inmunoenzimas , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas de Neoplasias/metabolismo , Reacción en Cadena de la Polimerasa , ReproducciónRESUMEN
Mastectomy in the guinea pig increases the incidence of still births and neonatal deaths compared with intact animals. The guinea pig has the endometrial gland cells (EGC) as the major source of relaxin, and this hormone has possible roles in uterine quiescence, cervical dilatation, and lengthening of the interpubic ligament in pregnancy. An effect of mastectomy on uterine relaxin has been sought by immunocytochemical localization during the estrous cycle, mid and late pregnancy and on endometrial relaxin gene expression in the late pregnant mastectomized animal by Northern analysis. Endometrium from midpregnant (day 35) and late pregnant (day 63) and from lactating (days 5, 21, and 28) guinea pigs immunostained with antiserum to porcine relaxin by the avidinbiotin technique. By increasing the sensitivity of the latter, relaxin immunostaining was also detected for the first time in EGC from cyclic animals (days 9 and 14). A pattern and intensity of relaxin immunostaining could be readily assigned to each of the stages examined: estrous cycle, midpregnancy, late pregnancy, lactation, and post weaning. The dark uniform staining of the EGC in late pregnancy was followed by sporadic staining of the EGC in lactation, returning to the cyclic picture after weaning. Endometrium from mastectomized cyclic and late pregnant guinea pigs showed a reduction in the amount of immunostaining compared with the relevant control animals. The reduction was most pronounced in the mastectomized late pregnant guinea pig. This result was reflected in an apparently lower level of relaxin mRNA in the endometrium of these animals compared to intact controls. These data indicate a novel linkage between the mammary gland, either directly or indirectly, to the nonpregnant or pregnant uterus. The loss of this signal appears to be associated with subsequent problems at parturition which may be linked to the reduction in endometrial gland relaxin production. The nature of this signal from the mammary gland, normally considered to be an exocrine rather than an endocrine gland, is unknown.
Asunto(s)
Endometrio/metabolismo , Estro/metabolismo , Mastectomía , Preñez/metabolismo , Relaxina/metabolismo , Animales , Northern Blotting , Densitometría , Femenino , Cobayas , Inmunohistoquímica , Lactancia/metabolismo , Concentración Osmolar , Embarazo , ARN Mensajero/metabolismo , Relaxina/genética , Transcripción GenéticaRESUMEN
Northern analysis was used to identify relaxin gene expression in ovaries of prepubertal pigs primed with pregnant mare's serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG). The cellular distribution of relaxin transcript in the developing follicle was localized by in-situ hybridization histochemistry. Three probes complementary to non-overlapping regions of the porcine prorelaxin molecule were used to identify relaxin gene expression in ovarian follicular tissue collected 0, 48, 60, 72 and 84 h after treatment with PMSG/hCG. A 1 kb transcript was detected in ovarian extracts of prepubertal gilts from 48 to 84 h after PMSG stimulation. This corresponds to the molecular size of the relaxin transcript reported in the pregnant sow ovary. Relaxin mRNA levels increased in ovaries from animals 48 through 84 h after PMSG. In-situ hybridization showed that the site of relaxin synthesis was the theca interna layer of the developing follicle. Relaxin mRNA was not observed in other follicular cell types, in small or atretic follicles or in follicles from unstimulated animals. The distribution and relative concentration of relaxin mRNA showed a good correlation with in-vitro production and immunohistochemical localization of relaxin previously reported in the developing pig follicle. The presence of both protein and mRNA for relaxin in the growing follicle supports a role for relaxin as a local regulator of ovarian function.
Asunto(s)
Gonadotropina Coriónica/farmacología , Regulación de la Expresión Génica/fisiología , Gonadotropinas Equinas/farmacología , Folículo Ovárico/fisiología , ARN Mensajero/análisis , Relaxina/genética , Animales , Secuencia de Bases , Northern Blotting , Sondas de ADN , Combinación de Medicamentos , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos , Folículo Ovárico/efectos de los fármacos , Relaxina/análisis , Porcinos/fisiología , Células Tecales/efectos de los fármacos , Células Tecales/fisiologíaRESUMEN
The ovaries of the ascidian Herdmania momus were extracted for relaxin. Relaxin immunoactivity was eluted from Sephadex G-50 in the position of authentic porcine ovarian relaxin and these fractions were parallel to porcine relaxin in an homologous porcine relaxin radioimmunoassay. Poly(A)+ RNA from ascidian ovaries hybridized to a 32P-labeled porcine relaxin B-chain 48 mer oligonucleotide probe. There was no hybridization with 32P-labeled probes of equal length but was rather directed to the B-chain of human relaxin or to two different regions of the C-peptide of porcine relaxin. We conclude that the ascidian ovary produces a relaxin-like molecule, possibly with greater homology in the B-chain to mammalian relaxins than in the C-peptide region.
Asunto(s)
Ovario/metabolismo , ARN Mensajero/análisis , Relaxina/biosíntesis , Urocordados/metabolismo , Animales , Secuencia de Bases , Northern Blotting , Cromatografía por Intercambio Iónico , Femenino , Datos de Secuencia Molecular , Sondas ARN , ARN Mensajero/aislamiento & purificación , Radioinmunoensayo , Relaxina/aislamiento & purificaciónRESUMEN
Relaxin (RLX) mRNA was sought in the corpus luteum of the pig during the cycle, pregnancy, and lactation using Northern analysis and in situ hybridization. Three oligonucleotide probes to regions of the preprorelaxin molecule were used for hybridization to detect RLX message and gave similar results. Northern analysis showed a single 1-kilobase RLX transcript at all three stages of the reproductive cycle studied. The intensity of the RLX hybridization signal was greatest in pregnancy and varied during the cycle. The signal in ovaries from day 3 cyclic animals increased by day 13 and declined on day 19. However, the hybridization signal at midcycle was only 2% of that during pregnancy. After parturition on day 2 of lactation, RLX message was still detected in the ovary, although at reduced levels compared with those during the cycle and pregnancy. In situ hybridization results showed hybridization to RLX mRNA in luteal tissue on day 13 of the cycle, but not on day 3 or 19. An increased hybridization signal was observed on days 40, 60, and 90 of pregnancy, with a decline on day 2 of lactation. Control sections incubated with labeled heterologous probe or preincubated with excess unlabeled probe did not hybridize. These results indicate a good correlation between the relative concentrations of RLX transcript and immunohistochemical results previously reported in the corpus luteum of the sow. In addition, they demonstrate that the RLX gene is expressed in luteal tissue, not only in pregnancy, but also in the cycle and early lactation.
Asunto(s)
Cuerpo Lúteo/metabolismo , Estro/metabolismo , Expresión Génica , Lactancia/metabolismo , Preñez/metabolismo , Relaxina/genética , Animales , Secuencia de Bases , Femenino , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos , Embarazo , ARN Mensajero/análisis , PorcinosRESUMEN
Relaxin is a product of the endometrial gland cells in the guinea pig. Mammary tissue was collected from intact cyclic animals, on days 35 and 63 of pregnancy, days 5 and 21 of lactation, and day 28 postpartum. Tissue was collected from six cyclic hysterectomized animals. Sections of mammary gland were immunostained with the avidin-biotin immunoperoxidase method, and antisera to porcine relaxin which were raised to different preparations in different laboratories. Light uniform staining was evident in the cytoplasm of all of the cuboidal epithelial cells forming the mammary duct system in cyclic animals; mammary gland from hysterectomized animals showed similar staining. At midpregnancy light staining was seen in some epithelial cells, and by late pregnancy there was only faint staining. On day 5 of lactation there was intense and uniform staining throughout the epithelial cells of the alveoli. By day 21 of lactation the cells still immunostained for relaxin, but on day 28 postpartum there was a return to the cyclic staining pattern. Endometrium from animals at 55-60 days of pregnancy and mammary gland from day 6 of lactation were used for poly(A)+ RNA isolation and Northern analysis. Three 48-mer oligonucleotide probes were used. Poly(A)+ RNA from endometrium of late pregnant guinea pigs and from the mammary gland in lactation hybridized with the same two probes and failed to hybridize with a third under moderate stringency conditions. The results suggest that the major source of relaxin in the guinea pig is sequential, the pregnant uterus and the lactating mammary gland. The local and/or systemic significance is not known.
Asunto(s)
Glándulas Mamarias Animales/metabolismo , Relaxina/metabolismo , Animales , Northern Blotting , Femenino , Cobayas , Inmunohistoquímica , Glándulas Mamarias Animales/citología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Relaxina/genéticaRESUMEN
The endometrium of late pregnant guinea-pigs was found to contain mRNA for relaxin. Poly(A)+RNA hybridized to 32P-labelled porcine relaxin-specific oligonucleotide probes. These probes corresponded to the C-peptide region of the prohormone. There was no hybridization to a 32P-labelled probe to the B-chain of porcine relaxin even under conditions of low stringency. The size of the relaxin mRNA was approximately 1.0 kb and similar to that found for relaxin mRNA from the pregnant sow ovary. This is the first study on relaxin mRNA from a uterine source.