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MHC class I genes transcribed in the murine placenta
A.50
Placenta
MHC Class I Genes Transcribed in the Murine Placenta. C.P. Landel’, D.L. Stabley*, E.N. Colyer*, C.A. Barone*, and S.L. Sipest, *Department of Clinical Science, Alfred I. duPont Institute, Wilmington, DE and tDepartment of Obstetrics and Gynecology, Yale School of Medicine, New Haven, CT. We have previously identified a novel murine MHC Class lb gene, Blastocyst MHC, that is expressed in the placenta of laboratory mice. We have also shown that, although this gene has been evolutionarily conserved in the mouse species M. spretus, BALBlc strains lack this gene. The phenotypic effect of this gene deletion is subtle; this implies that other genes might exist that act in parallel with Blastocyst MHC. We therefore set out to identify other MHC class lb genes that are also expressed in the murine placenta. We used an RTPCR based strategy to amplify, clone and sequence MHC class I genes transcribed in the murine placenta. We have identified a number of class I genes expressed in this tissue, including four known class lb genes for which transcripts have not previously been reported, as well as several novel class lb gene transcripts. We postulate that this spectrum of placental-specific MHC class lb genes act together at the maternal-fetal interface in ways that are important for a successful pregnancy,
Phospholipase Gestational G. Rice, Women’s
A, lsozyme Expression
in Human Term
Tissues. K. Freed, E. Moses, SBrennecke and Department of Perinatal Medicine, Royal Hospital, Carlton. Victoria, Australia, 3053.
NUMBERS OF ENDOMETRIAL NATURAL KiLL,ER CELLS RELATIVE TO CONCEPTUS ATTACHMENT SITESDURING EARLY PREGNANCYIN THE PIG. H Enrrelhardt’**, B. A. Croy’ & G. J. King’, Depts. of ‘Animal & Poultry Science, and ‘Biomedical Sciences, University of Guelph, Guelph, Ontario, CANADA. Uterine natural killer (NK)-like lytic activity is elevated around the time of conceptusattachment in the pig (dlO20; term = 115 days). In this study, uterine NK cell numbers at (WA) versus between (W) attachment sites during early pregnancy were compared using flow cytometry. Endometrial tissue was dispersedenzymatically, enriched for leukocytes by gradient centrifugation and stained with either G7 (binds to porcine NK cells, granulocytes and macrophages/monocytes(Mb); gift of Y .B. Kim, Univ. of Chicago) or CD44 @an leukocyte marker) followed by goat antimouse-FITC. G7 binding was similar at d1.5 of the cycle (n=8), and d16 (n=3) and d22 (n = 10) of pregnancy but was elevated at UtA versus UtB by d26 of gestation (37.3+5.5% vs. 14.4*5.0%, resp.; n-4). However, expressing G7 binding as a percentage of CD44+ cells removed this difference. G7 binding was higher in uterine cells than in peripheral blood cells (PBMC; 43.2&2.4% vs. 29.0f3.3%, resp.). Detailed analysisof scatter characteristicsrevealed that G7 stained similar numbers of M4 and lymphocytes in PBMC, while the main G7+ cell type in uterus was a large granular lymphocyte (3-10 fold more frequent than M4). These findings suggest that, as in humans and rodents, large granular lymphocytes with NK-like activity are a feature of pregnancy in the pig, a species with non-invasive, epitheliochorial placentatlon. Supported by NSERC and OMAFRA
Distribution Messenger
of the Phospholipase A, RNA in Human Term Gestational
E. Moses, K. Freed,G. of Perinatal Medicine,
Receptor Tissues.
Rice and S. Brennecke, Department Royal Women’s Hospital, Cerlton,
Victoria, Australia, 3053. The aim of this study was to establish the tissue-specific expression of mRNAtranscripts encoding Type II, Type IV and cytosolic phospholipase A, [PLA,] isozymes in human
term amnion, choriodecida
and placenta.
Gestational
tissues [n=3) were collected at elective Caesarean section at 3741 weeks of gestation. Total RNA was isolated by guenidinium thiocyenante phenol-chloroform extraction. PolyA’mRNA was isolated using the PolyATract mRNA isolation System 111 [Promegal. Relative mRNAexpression was determined by Northern blot analysis and laser densitometry, using an 800 bp
and 400 bp cDNA for human Type II and Type IV PI-A,, respectively, and a 2.5kb cDNAfor human cytosolic PLA,. Data were normalised against oligo[dT] probe hybridisation. Type II end IV PLA, mRNA transcripts were most abundantly expressed in placenta [p < 0.01 I. In contrast, cytosolic PLA, mRNA was greater in amnion than in choriodecidua or placenta [p< 0.01 I. These data are consistent with the hypothesis that secretory PLA, isozymes contribute to phospholipid metabolism and second messenger formation to a greater extent in the placenta than in fetal membranes, while cytosolic PLA, represents the major isozyme in human amnion.
The aim of this study was to determine,
in human term
gestational tissues, the mRNA expression profilefor a recently cloned human PLA, receptor. PLA,-receptor mRNA was detected in amnion, choriodecidua end placenta by RT-PCR and transcripts corresponding in size to the 6.5 kb and 5.4 kb transcripts previously reported in ofher human tissues, were detected in choriodecidua by Northern blot hybridization. Smaller
transcripts of ca. 2.3 kb and ca. 1.2 kb were detected by Northern blot hybridization in amnion and placenta, respectively. Alternative polyadenylation accounts for the 6.5 kb and 5.4 kb transcripts,
which
encode
membrane-
bound and secreted receptor forms, respectively. The novel smaller transcripts detected in amnion and placenta await. further cheracterisation. The presence of PLA, receptor mRNA indicates that an alternative noncatalytic pathway may contribute to the effects of secretory PlAz isozymes in these tissues. PLA, isozymes released from human gestational tissues during pregnancy and at the time of labour may also function as paracrine or eutocrine mediators to affect cell function.
(1996),
Vol. 17
Placenta
MHC Class I Genes Transcribed in the Murine Placenta. C.P. Landel’, D.L. Stabley*, E.N. Colyer*, C.A. Barone*, and S.L. Sipest, *Department of Clinical Science, Alfred I. duPont Institute, Wilmington, DE and tDepartment of Obstetrics and Gynecology, Yale School of Medicine, New Haven, CT. We have previously identified a novel murine MHC Class lb gene, Blastocyst MHC, that is expressed in the placenta of laboratory mice. We have also shown that, although this gene has been evolutionarily conserved in the mouse species M. spretus, BALBlc strains lack this gene. The phenotypic effect of this gene deletion is subtle; this implies that other genes might exist that act in parallel with Blastocyst MHC. We therefore set out to identify other MHC class lb genes that are also expressed in the murine placenta. We used an RTPCR based strategy to amplify, clone and sequence MHC class I genes transcribed in the murine placenta. We have identified a number of class I genes expressed in this tissue, including four known class lb genes for which transcripts have not previously been reported, as well as several novel class lb gene transcripts. We postulate that this spectrum of placental-specific MHC class lb genes act together at the maternal-fetal interface in ways that are important for a successful pregnancy,
Phospholipase Gestational G. Rice, Women’s
A, lsozyme Expression
in Human Term
Tissues. K. Freed, E. Moses, SBrennecke and Department of Perinatal Medicine, Royal Hospital, Carlton. Victoria, Australia, 3053.
NUMBERS OF ENDOMETRIAL NATURAL KiLL,ER CELLS RELATIVE TO CONCEPTUS ATTACHMENT SITESDURING EARLY PREGNANCYIN THE PIG. H Enrrelhardt’**, B. A. Croy’ & G. J. King’, Depts. of ‘Animal & Poultry Science, and ‘Biomedical Sciences, University of Guelph, Guelph, Ontario, CANADA. Uterine natural killer (NK)-like lytic activity is elevated around the time of conceptusattachment in the pig (dlO20; term = 115 days). In this study, uterine NK cell numbers at (WA) versus between (W) attachment sites during early pregnancy were compared using flow cytometry. Endometrial tissue was dispersedenzymatically, enriched for leukocytes by gradient centrifugation and stained with either G7 (binds to porcine NK cells, granulocytes and macrophages/monocytes(Mb); gift of Y .B. Kim, Univ. of Chicago) or CD44 @an leukocyte marker) followed by goat antimouse-FITC. G7 binding was similar at d1.5 of the cycle (n=8), and d16 (n=3) and d22 (n = 10) of pregnancy but was elevated at UtA versus UtB by d26 of gestation (37.3+5.5% vs. 14.4*5.0%, resp.; n-4). However, expressing G7 binding as a percentage of CD44+ cells removed this difference. G7 binding was higher in uterine cells than in peripheral blood cells (PBMC; 43.2&2.4% vs. 29.0f3.3%, resp.). Detailed analysisof scatter characteristicsrevealed that G7 stained similar numbers of M4 and lymphocytes in PBMC, while the main G7+ cell type in uterus was a large granular lymphocyte (3-10 fold more frequent than M4). These findings suggest that, as in humans and rodents, large granular lymphocytes with NK-like activity are a feature of pregnancy in the pig, a species with non-invasive, epitheliochorial placentatlon. Supported by NSERC and OMAFRA
Distribution Messenger
of the Phospholipase A, RNA in Human Term Gestational
E. Moses, K. Freed,G. of Perinatal Medicine,
Receptor Tissues.
Rice and S. Brennecke, Department Royal Women’s Hospital, Cerlton,
Victoria, Australia, 3053. The aim of this study was to establish the tissue-specific expression of mRNAtranscripts encoding Type II, Type IV and cytosolic phospholipase A, [PLA,] isozymes in human
term amnion, choriodecida
and placenta.
Gestational
tissues [n=3) were collected at elective Caesarean section at 3741 weeks of gestation. Total RNA was isolated by guenidinium thiocyenante phenol-chloroform extraction. PolyA’mRNA was isolated using the PolyATract mRNA isolation System 111 [Promegal. Relative mRNAexpression was determined by Northern blot analysis and laser densitometry, using an 800 bp
and 400 bp cDNA for human Type II and Type IV PI-A,, respectively, and a 2.5kb cDNAfor human cytosolic PLA,. Data were normalised against oligo[dT] probe hybridisation. Type II end IV PLA, mRNA transcripts were most abundantly expressed in placenta [p < 0.01 I. In contrast, cytosolic PLA, mRNA was greater in amnion than in choriodecidua or placenta [p< 0.01 I. These data are consistent with the hypothesis that secretory PLA, isozymes contribute to phospholipid metabolism and second messenger formation to a greater extent in the placenta than in fetal membranes, while cytosolic PLA, represents the major isozyme in human amnion.
The aim of this study was to determine,
in human term
gestational tissues, the mRNA expression profilefor a recently cloned human PLA, receptor. PLA,-receptor mRNA was detected in amnion, choriodecidua end placenta by RT-PCR and transcripts corresponding in size to the 6.5 kb and 5.4 kb transcripts previously reported in ofher human tissues, were detected in choriodecidua by Northern blot hybridization. Smaller
transcripts of ca. 2.3 kb and ca. 1.2 kb were detected by Northern blot hybridization in amnion and placenta, respectively. Alternative polyadenylation accounts for the 6.5 kb and 5.4 kb transcripts,
which
encode
membrane-
bound and secreted receptor forms, respectively. The novel smaller transcripts detected in amnion and placenta await. further cheracterisation. The presence of PLA, receptor mRNA indicates that an alternative noncatalytic pathway may contribute to the effects of secretory PlAz isozymes in these tissues. PLA, isozymes released from human gestational tissues during pregnancy and at the time of labour may also function as paracrine or eutocrine mediators to affect cell function.
(1996),
Vol. 17