Identification of Genes Enriched in GnRH Neurons by Translating Ribosome Affinity Purification and RNAseq in Mice
Gonadotropin-releasing hormone (GnRH) neurons are a nexus of fertility regulation. We used translating ribosome affinity purification coupled with RNA sequencing to examine messenger RNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice. GnRH neuron ribosomes were tagged...
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| Veröffentlicht in: | Endocrinology (Philadelphia) 2018-04, Vol.159 (4), p.1922-1940 |
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| container_title | Endocrinology (Philadelphia) |
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| creator | Burger, Laura L Vanacker, Charlotte Phumsatitpong, Chayarndorn Wagenmaker, Elizabeth R Wang, Luhong Olson, David P Moenter, Suzanne M |
| description | Gonadotropin-releasing hormone (GnRH) neurons are a nexus of fertility regulation. We used translating ribosome affinity purification coupled with RNA sequencing to examine messenger RNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice. GnRH neuron ribosomes were tagged with green fluorescent protein (GFP) and GFP-labeled polysomes isolated by immunoprecipitation, producing one RNA fraction enhanced for GnRH neuron transcripts and one RNA fraction depleted. Complementary DNA libraries were created from each fraction and 50-base, paired-end sequencing done and differential expression (enhanced fraction/depleted fraction) determined with a threshold of >1.5- or |
| doi_str_mv | 10.1210/en.2018-00001 |
| format | Article |
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We used translating ribosome affinity purification coupled with RNA sequencing to examine messenger RNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice. GnRH neuron ribosomes were tagged with green fluorescent protein (GFP) and GFP-labeled polysomes isolated by immunoprecipitation, producing one RNA fraction enhanced for GnRH neuron transcripts and one RNA fraction depleted. Complementary DNA libraries were created from each fraction and 50-base, paired-end sequencing done and differential expression (enhanced fraction/depleted fraction) determined with a threshold of >1.5- or <0.66-fold (false discovery rate P ≤ 0.05). A core of ∼840 genes was differentially expressed in GnRH neurons in all treatments, including enrichment for Gnrh1 (∼40-fold), and genes critical for GnRH neuron and/or gonadotrope development. In contrast, non-neuronal transcripts were not enriched or were de-enriched. Several epithelial markers were also enriched, consistent with the olfactory epithelial origins of GnRH neurons. Interestingly, many synaptic transmission pathways were de-enriched, in accordance with relatively low innervation of GnRH neurons. The most striking difference between intact and GDX mice of both sexes was a marked downregulation of genes associated with oxidative phosphorylation and upregulation of glucose transporters in GnRH neurons from GDX mice. This may suggest that GnRH neurons switch to an alternate fuel to increase adenosine triphosphate production in the absence of negative feedback when GnRH release is elevated. Knowledge of the GnRH neuron translatome and its regulation can guide functional studies and can be extended to disease states, such as polycystic ovary syndrome.</description><identifier>ISSN: 1945-7170</identifier><identifier>ISSN: 0013-7227</identifier><identifier>EISSN: 1945-7170</identifier><identifier>DOI: 10.1210/en.2018-00001</identifier><identifier>PMID: 29522155</identifier><language>eng</language><publisher>United States: Endocrine Society</publisher><subject>Animals ; Female ; Gene Expression ; Gonadotropin-Releasing Hormone - metabolism ; Green Fluorescent Proteins - metabolism ; Hypothalamus - metabolism ; Male ; Mice ; Neurons - metabolism ; Sequence Analysis, RNA</subject><ispartof>Endocrinology (Philadelphia), 2018-04, Vol.159 (4), p.1922-1940</ispartof><rights>Copyright © 2018 Endocrine Society 2018</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c387t-505be91178633674ad591c447022d814848fe0bf2c55b8dc82cfeaba4b2747233</citedby><cites>FETCH-LOGICAL-c387t-505be91178633674ad591c447022d814848fe0bf2c55b8dc82cfeaba4b2747233</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29522155$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Burger, Laura L</creatorcontrib><creatorcontrib>Vanacker, Charlotte</creatorcontrib><creatorcontrib>Phumsatitpong, Chayarndorn</creatorcontrib><creatorcontrib>Wagenmaker, Elizabeth R</creatorcontrib><creatorcontrib>Wang, Luhong</creatorcontrib><creatorcontrib>Olson, David P</creatorcontrib><creatorcontrib>Moenter, Suzanne M</creatorcontrib><title>Identification of Genes Enriched in GnRH Neurons by Translating Ribosome Affinity Purification and RNAseq in Mice</title><title>Endocrinology (Philadelphia)</title><addtitle>Endocrinology</addtitle><description>Gonadotropin-releasing hormone (GnRH) neurons are a nexus of fertility regulation. We used translating ribosome affinity purification coupled with RNA sequencing to examine messenger RNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice. GnRH neuron ribosomes were tagged with green fluorescent protein (GFP) and GFP-labeled polysomes isolated by immunoprecipitation, producing one RNA fraction enhanced for GnRH neuron transcripts and one RNA fraction depleted. Complementary DNA libraries were created from each fraction and 50-base, paired-end sequencing done and differential expression (enhanced fraction/depleted fraction) determined with a threshold of >1.5- or <0.66-fold (false discovery rate P ≤ 0.05). A core of ∼840 genes was differentially expressed in GnRH neurons in all treatments, including enrichment for Gnrh1 (∼40-fold), and genes critical for GnRH neuron and/or gonadotrope development. In contrast, non-neuronal transcripts were not enriched or were de-enriched. Several epithelial markers were also enriched, consistent with the olfactory epithelial origins of GnRH neurons. Interestingly, many synaptic transmission pathways were de-enriched, in accordance with relatively low innervation of GnRH neurons. The most striking difference between intact and GDX mice of both sexes was a marked downregulation of genes associated with oxidative phosphorylation and upregulation of glucose transporters in GnRH neurons from GDX mice. This may suggest that GnRH neurons switch to an alternate fuel to increase adenosine triphosphate production in the absence of negative feedback when GnRH release is elevated. Knowledge of the GnRH neuron translatome and its regulation can guide functional studies and can be extended to disease states, such as polycystic ovary syndrome.</description><subject>Animals</subject><subject>Female</subject><subject>Gene Expression</subject><subject>Gonadotropin-Releasing Hormone - metabolism</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Hypothalamus - metabolism</subject><subject>Male</subject><subject>Mice</subject><subject>Neurons - metabolism</subject><subject>Sequence Analysis, RNA</subject><issn>1945-7170</issn><issn>0013-7227</issn><issn>1945-7170</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkUtPAyEURonR-F66NSzdjAIDhdmYNEariVbT6JowzEUxU7AwY9J_79T6ZAMJh3O590PoiJJTyig5g3DKCFUFGRbdQLu04qKQVJLNP-cdtJfz6wBwzstttMMqwRgVYhctbhoInXfems7HgKPDEwiQ8WVI3r5Ag33AkzC7xlPoUwwZ10v8mEzI7fAgPOOZr2OOc8Bj53zw3RI_9OnXZ0KDZ9NxhsVKdOctHKAtZ9oMh1_7Pnq6uny8uC5u7yc3F-PbwpZKdoUgooaKUqlGZTmS3DSiopZzSRhrFOWKKwekdswKUavGKmYdmNrwmkkuWVnuo_O1962v59DYoc1kWv2W_NykpY7G6_83wb_o5_iuR0xJUbFBcPIlSHHRQ-703GcLbWsCxD7rYeysohVRYkCLNWpTzDmB-ylDiV7FpCGseKU_Yxr4479_-6G_cyk_ABb4joE</recordid><startdate>20180401</startdate><enddate>20180401</enddate><creator>Burger, Laura L</creator><creator>Vanacker, Charlotte</creator><creator>Phumsatitpong, Chayarndorn</creator><creator>Wagenmaker, Elizabeth R</creator><creator>Wang, Luhong</creator><creator>Olson, David P</creator><creator>Moenter, Suzanne M</creator><general>Endocrine Society</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20180401</creationdate><title>Identification of Genes Enriched in GnRH Neurons by Translating Ribosome Affinity Purification and RNAseq in Mice</title><author>Burger, Laura L ; Vanacker, Charlotte ; Phumsatitpong, Chayarndorn ; Wagenmaker, Elizabeth R ; Wang, Luhong ; Olson, David P ; Moenter, Suzanne M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c387t-505be91178633674ad591c447022d814848fe0bf2c55b8dc82cfeaba4b2747233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Animals</topic><topic>Female</topic><topic>Gene Expression</topic><topic>Gonadotropin-Releasing Hormone - metabolism</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Hypothalamus - metabolism</topic><topic>Male</topic><topic>Mice</topic><topic>Neurons - metabolism</topic><topic>Sequence Analysis, RNA</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Burger, Laura L</creatorcontrib><creatorcontrib>Vanacker, Charlotte</creatorcontrib><creatorcontrib>Phumsatitpong, Chayarndorn</creatorcontrib><creatorcontrib>Wagenmaker, Elizabeth R</creatorcontrib><creatorcontrib>Wang, Luhong</creatorcontrib><creatorcontrib>Olson, David P</creatorcontrib><creatorcontrib>Moenter, Suzanne M</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Endocrinology (Philadelphia)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Burger, Laura L</au><au>Vanacker, Charlotte</au><au>Phumsatitpong, Chayarndorn</au><au>Wagenmaker, Elizabeth R</au><au>Wang, Luhong</au><au>Olson, David P</au><au>Moenter, Suzanne M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of Genes Enriched in GnRH Neurons by Translating Ribosome Affinity Purification and RNAseq in Mice</atitle><jtitle>Endocrinology (Philadelphia)</jtitle><addtitle>Endocrinology</addtitle><date>2018-04-01</date><risdate>2018</risdate><volume>159</volume><issue>4</issue><spage>1922</spage><epage>1940</epage><pages>1922-1940</pages><issn>1945-7170</issn><issn>0013-7227</issn><eissn>1945-7170</eissn><abstract>Gonadotropin-releasing hormone (GnRH) neurons are a nexus of fertility regulation. We used translating ribosome affinity purification coupled with RNA sequencing to examine messenger RNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice. GnRH neuron ribosomes were tagged with green fluorescent protein (GFP) and GFP-labeled polysomes isolated by immunoprecipitation, producing one RNA fraction enhanced for GnRH neuron transcripts and one RNA fraction depleted. Complementary DNA libraries were created from each fraction and 50-base, paired-end sequencing done and differential expression (enhanced fraction/depleted fraction) determined with a threshold of >1.5- or <0.66-fold (false discovery rate P ≤ 0.05). A core of ∼840 genes was differentially expressed in GnRH neurons in all treatments, including enrichment for Gnrh1 (∼40-fold), and genes critical for GnRH neuron and/or gonadotrope development. In contrast, non-neuronal transcripts were not enriched or were de-enriched. Several epithelial markers were also enriched, consistent with the olfactory epithelial origins of GnRH neurons. Interestingly, many synaptic transmission pathways were de-enriched, in accordance with relatively low innervation of GnRH neurons. The most striking difference between intact and GDX mice of both sexes was a marked downregulation of genes associated with oxidative phosphorylation and upregulation of glucose transporters in GnRH neurons from GDX mice. This may suggest that GnRH neurons switch to an alternate fuel to increase adenosine triphosphate production in the absence of negative feedback when GnRH release is elevated. Knowledge of the GnRH neuron translatome and its regulation can guide functional studies and can be extended to disease states, such as polycystic ovary syndrome.</abstract><cop>United States</cop><pub>Endocrine Society</pub><pmid>29522155</pmid><doi>10.1210/en.2018-00001</doi><tpages>19</tpages><oa>free_for_read</oa></addata></record> |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE; Free E-Journal (出版社公開部分のみ）; Alma/SFX Local Collection; Journals@Ovid Complete |
| subjects | Animals Female Gene Expression Gonadotropin-Releasing Hormone - metabolism Green Fluorescent Proteins - metabolism Hypothalamus - metabolism Male Mice Neurons - metabolism Sequence Analysis, RNA |
| title | Identification of Genes Enriched in GnRH Neurons by Translating Ribosome Affinity Purification and RNAseq in Mice |
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