Distribution of aromatase cytochrome P450 messenger ribonucleic acid in adult rhesus monkey brains
The conversion of androgens to estrogens by aromatase cytochrome P450 (P450arom) is an important step in the mechanism of androgen action in the brain. However, the distribution of P450arom mRNA in adult rhesus monkey brains has not been studied because specific probes have not been available. To ad...
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| Veröffentlicht in: | Biology of reproduction 1997-10, Vol.57 (4), p.772-777 |
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| creator | ABDELGADIR, S. E ROSELLI, C. E CHOATE, J. V. A RESKO, J. A |
| description | The conversion of androgens to estrogens by aromatase cytochrome P450 (P450arom) is an important step in the mechanism of
androgen action in the brain. However, the distribution of P450arom mRNA in adult rhesus monkey brains has not been studied
because specific probes have not been available. To address this deficit, we cloned and sequenced a 455-basepair segment of
the 5' coding region of the rhesus P450arom cDNA. Total RNA was extracted from a rhesus monkey placenta (Day 47 of gestation
and subjected to reverse transcriptase (RT) polymerase chain reaction (PCR) using consensus oligonucleotide primers selected
from published human and rat P450arom DNA sequences. The RT-PCR product was subcloned into a vector and sequenced. The monkey
P450arom cDNA was 97% identical to the human sequence but shared only 86% homology with the rat sequence. We then developed
a ribonuclease protection assay using a monkey P450arom cDNA and studied the distribution of P450arom mRNA in adult monkey
brains. This assay protected two RNA fragments, one 455 nucleotides (nt) in length and the second approximately 300 nt. The
relative distribution of P450arom mRNA (the 455-nt fragment) between brain areas of the adult (n = 3) was high in the bed
nucleus of the stria terminalis > medial preoptic/anterior hypothalamus > amygdala; intermediate in the medial basal hypothalamus
(infundibular nucleus, median eminence, ventromedial nucleus) > lateral preoptic/anterior hypothalamus; and low in the septum
> lateral-dorsal-medial hypothalamus. P450arom mRNA was undetectable in cingulate and parietal cortex, hippocampus, and cerebellum.
P450arom activity, as measured by the 3H2O assay, correlated well with the distribution of P450arom mRNA (the 455-nt protected
fragment; r = 0.9) in the same tissues. A shorter protected RNA fragment was found in the medial basal hypothalamus, the bed
nucleus of the stria terminalis, the amygdala, and the cingulate and parietal cortex but not in the other brain areas investigated.
Its presence did not correlate with aromatase activity in brain tissue. This study describes the development of a ribonuclease
protection assay using a monkey cDNA produced by RT-PCR and its usefulness for studying the distribution of P450arom mRNA
in brains of nonhuman primates. |
| doi_str_mv | 10.1095/biolreprod57.4.772 |
| format | Article |
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androgen action in the brain. However, the distribution of P450arom mRNA in adult rhesus monkey brains has not been studied
because specific probes have not been available. To address this deficit, we cloned and sequenced a 455-basepair segment of
the 5' coding region of the rhesus P450arom cDNA. Total RNA was extracted from a rhesus monkey placenta (Day 47 of gestation
and subjected to reverse transcriptase (RT) polymerase chain reaction (PCR) using consensus oligonucleotide primers selected
from published human and rat P450arom DNA sequences. The RT-PCR product was subcloned into a vector and sequenced. The monkey
P450arom cDNA was 97% identical to the human sequence but shared only 86% homology with the rat sequence. We then developed
a ribonuclease protection assay using a monkey P450arom cDNA and studied the distribution of P450arom mRNA in adult monkey
brains. This assay protected two RNA fragments, one 455 nucleotides (nt) in length and the second approximately 300 nt. The
relative distribution of P450arom mRNA (the 455-nt fragment) between brain areas of the adult (n = 3) was high in the bed
nucleus of the stria terminalis > medial preoptic/anterior hypothalamus > amygdala; intermediate in the medial basal hypothalamus
(infundibular nucleus, median eminence, ventromedial nucleus) > lateral preoptic/anterior hypothalamus; and low in the septum
> lateral-dorsal-medial hypothalamus. P450arom mRNA was undetectable in cingulate and parietal cortex, hippocampus, and cerebellum.
P450arom activity, as measured by the 3H2O assay, correlated well with the distribution of P450arom mRNA (the 455-nt protected
fragment; r = 0.9) in the same tissues. A shorter protected RNA fragment was found in the medial basal hypothalamus, the bed
nucleus of the stria terminalis, the amygdala, and the cingulate and parietal cortex but not in the other brain areas investigated.
Its presence did not correlate with aromatase activity in brain tissue. This study describes the development of a ribonuclease
protection assay using a monkey cDNA produced by RT-PCR and its usefulness for studying the distribution of P450arom mRNA
in brains of nonhuman primates.</description><identifier>ISSN: 0006-3363</identifier><identifier>EISSN: 1529-7268</identifier><identifier>DOI: 10.1095/biolreprod57.4.772</identifier><identifier>PMID: 9314579</identifier><identifier>CODEN: BIREBV</identifier><language>eng</language><publisher>Madison, WI: Society for the Study of Reproduction</publisher><subject>Animals ; Aromatase - metabolism ; Base Sequence ; Biochemistry and metabolism ; Biological and medical sciences ; Brain Chemistry - physiology ; Central nervous system ; Cloning, Molecular ; DNA Probes ; Fundamental and applied biological sciences. Psychology ; Humans ; Macaca mulatta ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Rats ; Ribonucleases - metabolism ; RNA, Messenger - analysis ; RNA, Messenger - isolation & purification ; RNA, Messenger - metabolism ; Vertebrates: nervous system and sense organs</subject><ispartof>Biology of reproduction, 1997-10, Vol.57 (4), p.772-777</ispartof><rights>1997 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2818126$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9314579$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>ABDELGADIR, S. E</creatorcontrib><creatorcontrib>ROSELLI, C. E</creatorcontrib><creatorcontrib>CHOATE, J. V. A</creatorcontrib><creatorcontrib>RESKO, J. A</creatorcontrib><title>Distribution of aromatase cytochrome P450 messenger ribonucleic acid in adult rhesus monkey brains</title><title>Biology of reproduction</title><addtitle>Biol Reprod</addtitle><description>The conversion of androgens to estrogens by aromatase cytochrome P450 (P450arom) is an important step in the mechanism of
androgen action in the brain. However, the distribution of P450arom mRNA in adult rhesus monkey brains has not been studied
because specific probes have not been available. To address this deficit, we cloned and sequenced a 455-basepair segment of
the 5' coding region of the rhesus P450arom cDNA. Total RNA was extracted from a rhesus monkey placenta (Day 47 of gestation
and subjected to reverse transcriptase (RT) polymerase chain reaction (PCR) using consensus oligonucleotide primers selected
from published human and rat P450arom DNA sequences. The RT-PCR product was subcloned into a vector and sequenced. The monkey
P450arom cDNA was 97% identical to the human sequence but shared only 86% homology with the rat sequence. We then developed
a ribonuclease protection assay using a monkey P450arom cDNA and studied the distribution of P450arom mRNA in adult monkey
brains. This assay protected two RNA fragments, one 455 nucleotides (nt) in length and the second approximately 300 nt. The
relative distribution of P450arom mRNA (the 455-nt fragment) between brain areas of the adult (n = 3) was high in the bed
nucleus of the stria terminalis > medial preoptic/anterior hypothalamus > amygdala; intermediate in the medial basal hypothalamus
(infundibular nucleus, median eminence, ventromedial nucleus) > lateral preoptic/anterior hypothalamus; and low in the septum
> lateral-dorsal-medial hypothalamus. P450arom mRNA was undetectable in cingulate and parietal cortex, hippocampus, and cerebellum.
P450arom activity, as measured by the 3H2O assay, correlated well with the distribution of P450arom mRNA (the 455-nt protected
fragment; r = 0.9) in the same tissues. A shorter protected RNA fragment was found in the medial basal hypothalamus, the bed
nucleus of the stria terminalis, the amygdala, and the cingulate and parietal cortex but not in the other brain areas investigated.
Its presence did not correlate with aromatase activity in brain tissue. This study describes the development of a ribonuclease
protection assay using a monkey cDNA produced by RT-PCR and its usefulness for studying the distribution of P450arom mRNA
in brains of nonhuman primates.</description><subject>Animals</subject><subject>Aromatase - metabolism</subject><subject>Base Sequence</subject><subject>Biochemistry and metabolism</subject><subject>Biological and medical sciences</subject><subject>Brain Chemistry - physiology</subject><subject>Central nervous system</subject><subject>Cloning, Molecular</subject><subject>DNA Probes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Macaca mulatta</subject><subject>Male</subject><subject>Molecular Sequence Data</subject><subject>Polymerase Chain Reaction</subject><subject>Rats</subject><subject>Ribonucleases - metabolism</subject><subject>RNA, Messenger - analysis</subject><subject>RNA, Messenger - isolation & purification</subject><subject>RNA, Messenger - metabolism</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0006-3363</issn><issn>1529-7268</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctqHDEQRUWwccZOfiAQ0MLxrid6q7UMzstgSBbJuik92q1E3XKkbob5-wg8eOtVUZzDhbqF0DtK9pQY-dHGnEp4LNlLvRd7rdkrtKOSmU4z1Z-hHSFEdZwr_hpd1vqHECo44xfownAqpDY7ZD_HupZotzXmBecRQ8kzrFADdsc1u6mtAf8UkuA51BqWh1Bw8_OyuRSiw-Cix3HB4Le04jKFulU85-VvOGJbIC71DTofIdXw9jSv0O-vX37dfu_uf3y7u_10303MqLXrnefGgxCEOa9H2RPNmDOC8YaJHx33vRqdDwSsMMwKDsz2DCRRzmjb8yt085TbCvm3hboOc6wupARLyFsddDtaKkVfFKliinElmvj-JG52Dn54LHGGchxO7TV-feJQHaSxwOJifdZYT3vKVNM-PGlTfJgOsYShzpBSC-XD4XCQehBD-x3_D62wjho</recordid><startdate>19971001</startdate><enddate>19971001</enddate><creator>ABDELGADIR, S. E</creator><creator>ROSELLI, C. E</creator><creator>CHOATE, J. V. A</creator><creator>RESKO, J. A</creator><general>Society for the Study of Reproduction</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19971001</creationdate><title>Distribution of aromatase cytochrome P450 messenger ribonucleic acid in adult rhesus monkey brains</title><author>ABDELGADIR, S. E ; ROSELLI, C. E ; CHOATE, J. V. A ; RESKO, J. A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h296t-8cd39da4402cd7f580722c94232960dfc3d86fcde0ab492b43a2b82a506c97b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Animals</topic><topic>Aromatase - metabolism</topic><topic>Base Sequence</topic><topic>Biochemistry and metabolism</topic><topic>Biological and medical sciences</topic><topic>Brain Chemistry - physiology</topic><topic>Central nervous system</topic><topic>Cloning, Molecular</topic><topic>DNA Probes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Macaca mulatta</topic><topic>Male</topic><topic>Molecular Sequence Data</topic><topic>Polymerase Chain Reaction</topic><topic>Rats</topic><topic>Ribonucleases - metabolism</topic><topic>RNA, Messenger - analysis</topic><topic>RNA, Messenger - isolation & purification</topic><topic>RNA, Messenger - metabolism</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>ABDELGADIR, S. E</creatorcontrib><creatorcontrib>ROSELLI, C. E</creatorcontrib><creatorcontrib>CHOATE, J. V. A</creatorcontrib><creatorcontrib>RESKO, J. A</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biology of reproduction</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>ABDELGADIR, S. E</au><au>ROSELLI, C. E</au><au>CHOATE, J. V. A</au><au>RESKO, J. A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Distribution of aromatase cytochrome P450 messenger ribonucleic acid in adult rhesus monkey brains</atitle><jtitle>Biology of reproduction</jtitle><addtitle>Biol Reprod</addtitle><date>1997-10-01</date><risdate>1997</risdate><volume>57</volume><issue>4</issue><spage>772</spage><epage>777</epage><pages>772-777</pages><issn>0006-3363</issn><eissn>1529-7268</eissn><coden>BIREBV</coden><abstract>The conversion of androgens to estrogens by aromatase cytochrome P450 (P450arom) is an important step in the mechanism of
androgen action in the brain. However, the distribution of P450arom mRNA in adult rhesus monkey brains has not been studied
because specific probes have not been available. To address this deficit, we cloned and sequenced a 455-basepair segment of
the 5' coding region of the rhesus P450arom cDNA. Total RNA was extracted from a rhesus monkey placenta (Day 47 of gestation
and subjected to reverse transcriptase (RT) polymerase chain reaction (PCR) using consensus oligonucleotide primers selected
from published human and rat P450arom DNA sequences. The RT-PCR product was subcloned into a vector and sequenced. The monkey
P450arom cDNA was 97% identical to the human sequence but shared only 86% homology with the rat sequence. We then developed
a ribonuclease protection assay using a monkey P450arom cDNA and studied the distribution of P450arom mRNA in adult monkey
brains. This assay protected two RNA fragments, one 455 nucleotides (nt) in length and the second approximately 300 nt. The
relative distribution of P450arom mRNA (the 455-nt fragment) between brain areas of the adult (n = 3) was high in the bed
nucleus of the stria terminalis > medial preoptic/anterior hypothalamus > amygdala; intermediate in the medial basal hypothalamus
(infundibular nucleus, median eminence, ventromedial nucleus) > lateral preoptic/anterior hypothalamus; and low in the septum
> lateral-dorsal-medial hypothalamus. P450arom mRNA was undetectable in cingulate and parietal cortex, hippocampus, and cerebellum.
P450arom activity, as measured by the 3H2O assay, correlated well with the distribution of P450arom mRNA (the 455-nt protected
fragment; r = 0.9) in the same tissues. A shorter protected RNA fragment was found in the medial basal hypothalamus, the bed
nucleus of the stria terminalis, the amygdala, and the cingulate and parietal cortex but not in the other brain areas investigated.
Its presence did not correlate with aromatase activity in brain tissue. This study describes the development of a ribonuclease
protection assay using a monkey cDNA produced by RT-PCR and its usefulness for studying the distribution of P450arom mRNA
in brains of nonhuman primates.</abstract><cop>Madison, WI</cop><pub>Society for the Study of Reproduction</pub><pmid>9314579</pmid><doi>10.1095/biolreprod57.4.772</doi><tpages>6</tpages></addata></record> |
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| ispartof | Biology of reproduction, 1997-10, Vol.57 (4), p.772-777 |
| issn | 0006-3363 1529-7268 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79315661 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Aromatase - metabolism Base Sequence Biochemistry and metabolism Biological and medical sciences Brain Chemistry - physiology Central nervous system Cloning, Molecular DNA Probes Fundamental and applied biological sciences. Psychology Humans Macaca mulatta Male Molecular Sequence Data Polymerase Chain Reaction Rats Ribonucleases - metabolism RNA, Messenger - analysis RNA, Messenger - isolation & purification RNA, Messenger - metabolism Vertebrates: nervous system and sense organs |
| title | Distribution of aromatase cytochrome P450 messenger ribonucleic acid in adult rhesus monkey brains |
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