Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig
Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential f...
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| Veröffentlicht in: | Biology of reproduction 1995-11, Vol.53 (5), p.1038-1050 |
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| creator | Geisert, R.D. (Oklahoma State University, Stillwater, OK.) Dixon, M.J Pratt, T Schmidt, R.A.M Lessley, B.A McCann, J.P |
| description | Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential for conceptus attachment. In the present study, a 30-kDa, basic endometrial glycoprotein (pGP30) was isolated and characterized during the estrous cycle and early pregnancy of the pig. Uterine flushings and endometrial culture media were obtained from gilts on Days 0, 5, 10, 12, 15, and 18 of the estrous cycle and Days 10, 12, 15, and 18 of pregnancy. A polyclonal antibody was generated to pGP30 after isolation of medium from Day 15 pregnant endometrial cultures separated by gel filtration and PAGE. Western blot analysis indicated that the antiserum reacted with isoforms of pGP30 and cross-reacted with a 90-kDa component in serum that was not removed after cleavage of the oligosaccharide chains from the 90-kDa glycoprotein. Antiserum did not detect a 30-kDa band in media from cultures of kidney, fat, heart, muscle, liver, or serum; however, heart and muscle did contain bands of different molecular masses that cross-reacted with the antiserum. Multiple bands of higher molecular mass (35-40 kDa) were detected in the endometrial cultures from gilts on Days 0 through 10 of the estrous cycle. Treatment of ovariectomized gilts with estradiol-17 beta stimulated a similar response. During the mid- to late luteal phase of the estrous cycle (Days 12-18), the 30-kDa band as well as an additional 32-kDa band was present on Western blots. Administration of progesterone for 14 days stimulated the synthesis of both the 30- and 32-kDa products in ovariectomized gilts. However, only the pGP30 was detected on Days 12-18 of pregnancy. Immunocytochemical localization with antiserum to pGP30 indicated that the glycoprotein is present in the endometrial epithelium |
| doi_str_mv | 10.1095/biolreprod53.5.1038 |
| format | Article |
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(Oklahoma State University, Stillwater, OK.) ; Dixon, M.J ; Pratt, T ; Schmidt, R.A.M ; Lessley, B.A ; McCann, J.P</creator><creatorcontrib>Geisert, R.D. (Oklahoma State University, Stillwater, OK.) ; Dixon, M.J ; Pratt, T ; Schmidt, R.A.M ; Lessley, B.A ; McCann, J.P</creatorcontrib><description>Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential for conceptus attachment. In the present study, a 30-kDa, basic endometrial glycoprotein (pGP30) was isolated and characterized during the estrous cycle and early pregnancy of the pig. Uterine flushings and endometrial culture media were obtained from gilts on Days 0, 5, 10, 12, 15, and 18 of the estrous cycle and Days 10, 12, 15, and 18 of pregnancy. A polyclonal antibody was generated to pGP30 after isolation of medium from Day 15 pregnant endometrial cultures separated by gel filtration and PAGE. Western blot analysis indicated that the antiserum reacted with isoforms of pGP30 and cross-reacted with a 90-kDa component in serum that was not removed after cleavage of the oligosaccharide chains from the 90-kDa glycoprotein. Antiserum did not detect a 30-kDa band in media from cultures of kidney, fat, heart, muscle, liver, or serum; however, heart and muscle did contain bands of different molecular masses that cross-reacted with the antiserum. Multiple bands of higher molecular mass (35-40 kDa) were detected in the endometrial cultures from gilts on Days 0 through 10 of the estrous cycle. Treatment of ovariectomized gilts with estradiol-17 beta stimulated a similar response. During the mid- to late luteal phase of the estrous cycle (Days 12-18), the 30-kDa band as well as an additional 32-kDa band was present on Western blots. Administration of progesterone for 14 days stimulated the synthesis of both the 30- and 32-kDa products in ovariectomized gilts. However, only the pGP30 was detected on Days 12-18 of pregnancy. Immunocytochemical localization with antiserum to pGP30 indicated that the glycoprotein is present in the endometrial epithelium</description><identifier>ISSN: 0006-3363</identifier><identifier>EISSN: 1529-7268</identifier><identifier>DOI: 10.1095/biolreprod53.5.1038</identifier><identifier>PMID: 8527506</identifier><identifier>CODEN: BIREBV</identifier><language>eng</language><publisher>Madison, WI: Society for the Study of Reproduction</publisher><subject>Animals ; Biological and medical sciences ; Blotting, Western ; CERDAS ; CHIMIE ; CICLO ESTRAL ; Culture Media, Conditioned - chemistry ; Culture Techniques ; CYCLE OESTRAL ; DESARROLLO EMBRIONARIO ; DEVELOPPEMENT EMBRYONNAIRE ; Endometrium - metabolism ; Epithelium - metabolism ; Estradiol - blood ; ESTROGENOS ; Estrus - physiology ; Female ; Fundamental and applied biological sciences. Psychology ; GESTACION ; GESTATION ; GLICOPROTEINAS ; GLYCOPROTEINE ; Glycoproteins - biosynthesis ; Glycoproteins - isolation & purification ; HORMONAS ; HORMONE ; Hormone metabolism and regulation ; Immunohistochemistry ; IMMUNOLOGIE ; INMUNOLOGIA ; Molecular Weight ; OESTROGENE ; Ovariectomy ; Pregnancy ; Pregnancy, Animal - physiology ; Pregnancy. Parturition. Lactation ; PROGESTERONA ; PROGESTERONE ; Progesterone - blood ; QUIMICA ; SINTESIS DE PROTEINAS ; Swine ; SYNTHESE PROTEIQUE ; Time Factors ; TRUIE ; UTERO ; UTERUS ; Vertebrates: reproduction</subject><ispartof>Biology of reproduction, 1995-11, Vol.53 (5), p.1038-1050</ispartof><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2897906$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8527506$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Geisert, R.D. (Oklahoma State University, Stillwater, OK.)</creatorcontrib><creatorcontrib>Dixon, M.J</creatorcontrib><creatorcontrib>Pratt, T</creatorcontrib><creatorcontrib>Schmidt, R.A.M</creatorcontrib><creatorcontrib>Lessley, B.A</creatorcontrib><creatorcontrib>McCann, J.P</creatorcontrib><title>Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig</title><title>Biology of reproduction</title><addtitle>Biol Reprod</addtitle><description>Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential for conceptus attachment. In the present study, a 30-kDa, basic endometrial glycoprotein (pGP30) was isolated and characterized during the estrous cycle and early pregnancy of the pig. Uterine flushings and endometrial culture media were obtained from gilts on Days 0, 5, 10, 12, 15, and 18 of the estrous cycle and Days 10, 12, 15, and 18 of pregnancy. A polyclonal antibody was generated to pGP30 after isolation of medium from Day 15 pregnant endometrial cultures separated by gel filtration and PAGE. Western blot analysis indicated that the antiserum reacted with isoforms of pGP30 and cross-reacted with a 90-kDa component in serum that was not removed after cleavage of the oligosaccharide chains from the 90-kDa glycoprotein. Antiserum did not detect a 30-kDa band in media from cultures of kidney, fat, heart, muscle, liver, or serum; however, heart and muscle did contain bands of different molecular masses that cross-reacted with the antiserum. Multiple bands of higher molecular mass (35-40 kDa) were detected in the endometrial cultures from gilts on Days 0 through 10 of the estrous cycle. Treatment of ovariectomized gilts with estradiol-17 beta stimulated a similar response. During the mid- to late luteal phase of the estrous cycle (Days 12-18), the 30-kDa band as well as an additional 32-kDa band was present on Western blots. Administration of progesterone for 14 days stimulated the synthesis of both the 30- and 32-kDa products in ovariectomized gilts. However, only the pGP30 was detected on Days 12-18 of pregnancy. Immunocytochemical localization with antiserum to pGP30 indicated that the glycoprotein is present in the endometrial epithelium</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>CERDAS</subject><subject>CHIMIE</subject><subject>CICLO ESTRAL</subject><subject>Culture Media, Conditioned - chemistry</subject><subject>Culture Techniques</subject><subject>CYCLE OESTRAL</subject><subject>DESARROLLO EMBRIONARIO</subject><subject>DEVELOPPEMENT EMBRYONNAIRE</subject><subject>Endometrium - metabolism</subject><subject>Epithelium - metabolism</subject><subject>Estradiol - blood</subject><subject>ESTROGENOS</subject><subject>Estrus - physiology</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GESTACION</subject><subject>GESTATION</subject><subject>GLICOPROTEINAS</subject><subject>GLYCOPROTEINE</subject><subject>Glycoproteins - biosynthesis</subject><subject>Glycoproteins - isolation & purification</subject><subject>HORMONAS</subject><subject>HORMONE</subject><subject>Hormone metabolism and regulation</subject><subject>Immunohistochemistry</subject><subject>IMMUNOLOGIE</subject><subject>INMUNOLOGIA</subject><subject>Molecular Weight</subject><subject>OESTROGENE</subject><subject>Ovariectomy</subject><subject>Pregnancy</subject><subject>Pregnancy, Animal - physiology</subject><subject>Pregnancy. Parturition. Lactation</subject><subject>PROGESTERONA</subject><subject>PROGESTERONE</subject><subject>Progesterone - blood</subject><subject>QUIMICA</subject><subject>SINTESIS DE PROTEINAS</subject><subject>Swine</subject><subject>SYNTHESE PROTEIQUE</subject><subject>Time Factors</subject><subject>TRUIE</subject><subject>UTERO</subject><subject>UTERUS</subject><subject>Vertebrates: reproduction</subject><issn>0006-3363</issn><issn>1529-7268</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkU1v1DAQhi0EKtvCHwAh-YB6S3Hs2ImPqHxVqsQBerYm_sgaHHuxs1qlf4C_jZddgS8jzzx6550ZhF635KYlkr8bfQrZ7nIynN3wmmPDE7RpOZVNT8XwFG0IIaJhTLDn6LKUH4S0HaPsAl0MnPaciA36fVdSgMWniCEarLeQQS82-8dTMjkMmJHm5wfANpo02yV7CHgKq0619WJ9xGWNy9YW_2gNNvvs44TrH9uy5LQvWK862L_yFnJY8S7bKULU61H9CO789AI9cxCKfXmOV-jh08fvt1-a-6-f727f3zeODnypszhwcmQjoX3rWpC67yXpDBsGTY10kjtnBJVUy0GMjnTaCKudk3SUhHYju0LXJ93q_de-OlSzL9qGANFWr6qvr6PdUME3Z3A_ztaoXfYz5FWdN1frb891KBqCy3UgX_5hdJDVmPjfb-un7cFnq8oMIVRRpg6HA2eKq-PhKvjqBDpICqZctR6-SdEK2kn2BzWal5E</recordid><startdate>19951101</startdate><enddate>19951101</enddate><creator>Geisert, R.D. (Oklahoma State University, Stillwater, OK.)</creator><creator>Dixon, M.J</creator><creator>Pratt, T</creator><creator>Schmidt, R.A.M</creator><creator>Lessley, B.A</creator><creator>McCann, J.P</creator><general>Society for the Study of Reproduction</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>19951101</creationdate><title>Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig</title><author>Geisert, R.D. (Oklahoma State University, Stillwater, OK.) ; Dixon, M.J ; Pratt, T ; Schmidt, R.A.M ; Lessley, B.A ; McCann, J.P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f285t-33faf9b3b0271f1a9c77904d388c2d9f95ffd6292c986bf04cd6ecff92b9024b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>CERDAS</topic><topic>CHIMIE</topic><topic>CICLO ESTRAL</topic><topic>Culture Media, Conditioned - chemistry</topic><topic>Culture Techniques</topic><topic>CYCLE OESTRAL</topic><topic>DESARROLLO EMBRIONARIO</topic><topic>DEVELOPPEMENT EMBRYONNAIRE</topic><topic>Endometrium - metabolism</topic><topic>Epithelium - metabolism</topic><topic>Estradiol - blood</topic><topic>ESTROGENOS</topic><topic>Estrus - physiology</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GESTACION</topic><topic>GESTATION</topic><topic>GLICOPROTEINAS</topic><topic>GLYCOPROTEINE</topic><topic>Glycoproteins - biosynthesis</topic><topic>Glycoproteins - isolation & purification</topic><topic>HORMONAS</topic><topic>HORMONE</topic><topic>Hormone metabolism and regulation</topic><topic>Immunohistochemistry</topic><topic>IMMUNOLOGIE</topic><topic>INMUNOLOGIA</topic><topic>Molecular Weight</topic><topic>OESTROGENE</topic><topic>Ovariectomy</topic><topic>Pregnancy</topic><topic>Pregnancy, Animal - physiology</topic><topic>Pregnancy. Parturition. Lactation</topic><topic>PROGESTERONA</topic><topic>PROGESTERONE</topic><topic>Progesterone - blood</topic><topic>QUIMICA</topic><topic>SINTESIS DE PROTEINAS</topic><topic>Swine</topic><topic>SYNTHESE PROTEIQUE</topic><topic>Time Factors</topic><topic>TRUIE</topic><topic>UTERO</topic><topic>UTERUS</topic><topic>Vertebrates: reproduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Geisert, R.D. (Oklahoma State University, Stillwater, OK.)</creatorcontrib><creatorcontrib>Dixon, M.J</creatorcontrib><creatorcontrib>Pratt, T</creatorcontrib><creatorcontrib>Schmidt, R.A.M</creatorcontrib><creatorcontrib>Lessley, B.A</creatorcontrib><creatorcontrib>McCann, J.P</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Biology of reproduction</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Geisert, R.D. (Oklahoma State University, Stillwater, OK.)</au><au>Dixon, M.J</au><au>Pratt, T</au><au>Schmidt, R.A.M</au><au>Lessley, B.A</au><au>McCann, J.P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig</atitle><jtitle>Biology of reproduction</jtitle><addtitle>Biol Reprod</addtitle><date>1995-11-01</date><risdate>1995</risdate><volume>53</volume><issue>5</issue><spage>1038</spage><epage>1050</epage><pages>1038-1050</pages><issn>0006-3363</issn><eissn>1529-7268</eissn><coden>BIREBV</coden><abstract>Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential for conceptus attachment. In the present study, a 30-kDa, basic endometrial glycoprotein (pGP30) was isolated and characterized during the estrous cycle and early pregnancy of the pig. Uterine flushings and endometrial culture media were obtained from gilts on Days 0, 5, 10, 12, 15, and 18 of the estrous cycle and Days 10, 12, 15, and 18 of pregnancy. A polyclonal antibody was generated to pGP30 after isolation of medium from Day 15 pregnant endometrial cultures separated by gel filtration and PAGE. Western blot analysis indicated that the antiserum reacted with isoforms of pGP30 and cross-reacted with a 90-kDa component in serum that was not removed after cleavage of the oligosaccharide chains from the 90-kDa glycoprotein. Antiserum did not detect a 30-kDa band in media from cultures of kidney, fat, heart, muscle, liver, or serum; however, heart and muscle did contain bands of different molecular masses that cross-reacted with the antiserum. Multiple bands of higher molecular mass (35-40 kDa) were detected in the endometrial cultures from gilts on Days 0 through 10 of the estrous cycle. Treatment of ovariectomized gilts with estradiol-17 beta stimulated a similar response. During the mid- to late luteal phase of the estrous cycle (Days 12-18), the 30-kDa band as well as an additional 32-kDa band was present on Western blots. Administration of progesterone for 14 days stimulated the synthesis of both the 30- and 32-kDa products in ovariectomized gilts. However, only the pGP30 was detected on Days 12-18 of pregnancy. Immunocytochemical localization with antiserum to pGP30 indicated that the glycoprotein is present in the endometrial epithelium</abstract><cop>Madison, WI</cop><pub>Society for the Study of Reproduction</pub><pmid>8527506</pmid><doi>10.1095/biolreprod53.5.1038</doi><tpages>13</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Biology of reproduction, 1995-11, Vol.53 (5), p.1038-1050 |
| issn | 0006-3363 1529-7268 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_77774248 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Biological and medical sciences Blotting, Western CERDAS CHIMIE CICLO ESTRAL Culture Media, Conditioned - chemistry Culture Techniques CYCLE OESTRAL DESARROLLO EMBRIONARIO DEVELOPPEMENT EMBRYONNAIRE Endometrium - metabolism Epithelium - metabolism Estradiol - blood ESTROGENOS Estrus - physiology Female Fundamental and applied biological sciences. Psychology GESTACION GESTATION GLICOPROTEINAS GLYCOPROTEINE Glycoproteins - biosynthesis Glycoproteins - isolation & purification HORMONAS HORMONE Hormone metabolism and regulation Immunohistochemistry IMMUNOLOGIE INMUNOLOGIA Molecular Weight OESTROGENE Ovariectomy Pregnancy Pregnancy, Animal - physiology Pregnancy. Parturition. Lactation PROGESTERONA PROGESTERONE Progesterone - blood QUIMICA SINTESIS DE PROTEINAS Swine SYNTHESE PROTEIQUE Time Factors TRUIE UTERO UTERUS Vertebrates: reproduction |
| title | Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig |
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