Identification of genes involved in urogenital development
<p>The mouse urogenital system comprises the gonads, mesonephroi, kidneys and adrenal glands. The indifferent gonad, or genital ridge, arises on the ventro-medial aspect of the mesonephros at approximately 10 days <em>post coitum</em> (dpc), and enlarges due to the influx of germ c...
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| Format: | Thesis |
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2002
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| _version_ | 1797087541128593408 |
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| author | Van Hateren, N |
| author_facet | Van Hateren, N |
| author_sort | Van Hateren, N |
| collection | OXFORD |
| description | <p>The mouse urogenital system comprises the gonads, mesonephroi, kidneys and adrenal glands. The indifferent gonad, or genital ridge, arises on the ventro-medial aspect of the mesonephros at approximately 10 days <em>post coitum</em> (dpc), and enlarges due to the influx of germ cells and migration of somatic cells from the coelomic epithelium and mesonephros. Male and female gonads are morphologically indistinguishable until 12.5 dpc when the expression of the testis-determining gene Sry in the gonadal supporting cell lineage results in the differentiation of Sertoli cells. This process initiates a cascade of gene expression leading to testis development and masculinisation of the embryo. In the absence of <em>Sry</em> the default ovarian pathway ensues.</p> <p>This project aims to identify genes exhibiting a spatio-temporal expression pattern consistent with a role in the development of the murine reproductive organs, with the primary focus on genes involved in Sertoli cell development. The main resource exploited was a normalised mouse urogenital ridge (NMUR) cDNA library constructed from gonad-mesonephros pairs at 11.5 dpc and 12.5 dpc. This library was screened by two methods: the first screen used sample sequencing of 472 NMUR cDNA clones to identify clones displaying homology to known developmentally important genes. The second screen employed DNA microarray technology to identify genes displaying differential expression in male and female gonads. The expression of candidate clones from both screens was examined by wholemount <em>in situ</em> hybridisation to identify clones displaying male-specific expression. These clones were studied in more detail to determine their cell-type specificity and timing of onset of expression.</p> <p>Two genes were found to display male-specific expression in Sertoli cells. One gene was the serine protease inhibitor, protease nexin 1 (<em>Pn1</em>); the other was a novel gene. The preliminary characterisation of this novel gene, termed Maestro (<em>Mro</em>), is presented.</p> <p><em>Pn1</em> and <em>Mro</em> are both expressed in a male-specific fashion prior to overt sexual differentiation. This timing, together with their expression in Sertoli cells, makes these genes candidates for regulation by the known sex-determining transcription factors (SRY, SOX9 etc.). In order to study the regulation of <em>Pn1</em> expression during gonad development, a <em>LacZ</em> reporter construct containing a genomic region immediately upstream of the <em>Pn1</em> gene was used to generate transgenic mice. This fragment did not drive Sertoli-cell specific expression of the reporter gene; therefore, the minimal gonadal enhancer for <em>Pn1</em> could not be identified.</p> |
| first_indexed | 2024-03-07T02:37:05Z |
| format | Thesis |
| id | oxford-uuid:a926f3b5-f968-4f85-9644-f4ad1e216fef |
| institution | University of Oxford |
| last_indexed | 2024-03-07T02:37:05Z |
| publishDate | 2002 |
| record_format | dspace |
| spelling | oxford-uuid:a926f3b5-f968-4f85-9644-f4ad1e216fef2022-03-27T03:06:28ZIdentification of genes involved in urogenital developmentThesishttp://purl.org/coar/resource_type/c_db06uuid:a926f3b5-f968-4f85-9644-f4ad1e216fefPolonsky Theses Digitisation Project2002Van Hateren, N<p>The mouse urogenital system comprises the gonads, mesonephroi, kidneys and adrenal glands. The indifferent gonad, or genital ridge, arises on the ventro-medial aspect of the mesonephros at approximately 10 days <em>post coitum</em> (dpc), and enlarges due to the influx of germ cells and migration of somatic cells from the coelomic epithelium and mesonephros. Male and female gonads are morphologically indistinguishable until 12.5 dpc when the expression of the testis-determining gene Sry in the gonadal supporting cell lineage results in the differentiation of Sertoli cells. This process initiates a cascade of gene expression leading to testis development and masculinisation of the embryo. In the absence of <em>Sry</em> the default ovarian pathway ensues.</p> <p>This project aims to identify genes exhibiting a spatio-temporal expression pattern consistent with a role in the development of the murine reproductive organs, with the primary focus on genes involved in Sertoli cell development. The main resource exploited was a normalised mouse urogenital ridge (NMUR) cDNA library constructed from gonad-mesonephros pairs at 11.5 dpc and 12.5 dpc. This library was screened by two methods: the first screen used sample sequencing of 472 NMUR cDNA clones to identify clones displaying homology to known developmentally important genes. The second screen employed DNA microarray technology to identify genes displaying differential expression in male and female gonads. The expression of candidate clones from both screens was examined by wholemount <em>in situ</em> hybridisation to identify clones displaying male-specific expression. These clones were studied in more detail to determine their cell-type specificity and timing of onset of expression.</p> <p>Two genes were found to display male-specific expression in Sertoli cells. One gene was the serine protease inhibitor, protease nexin 1 (<em>Pn1</em>); the other was a novel gene. The preliminary characterisation of this novel gene, termed Maestro (<em>Mro</em>), is presented.</p> <p><em>Pn1</em> and <em>Mro</em> are both expressed in a male-specific fashion prior to overt sexual differentiation. This timing, together with their expression in Sertoli cells, makes these genes candidates for regulation by the known sex-determining transcription factors (SRY, SOX9 etc.). In order to study the regulation of <em>Pn1</em> expression during gonad development, a <em>LacZ</em> reporter construct containing a genomic region immediately upstream of the <em>Pn1</em> gene was used to generate transgenic mice. This fragment did not drive Sertoli-cell specific expression of the reporter gene; therefore, the minimal gonadal enhancer for <em>Pn1</em> could not be identified.</p> |
| spellingShingle | Van Hateren, N Identification of genes involved in urogenital development |
| title | Identification of genes involved in urogenital development |
| title_full | Identification of genes involved in urogenital development |
| title_fullStr | Identification of genes involved in urogenital development |
| title_full_unstemmed | Identification of genes involved in urogenital development |
| title_short | Identification of genes involved in urogenital development |
| title_sort | identification of genes involved in urogenital development |
| work_keys_str_mv | AT vanhaterenn identificationofgenesinvolvedinurogenitaldevelopment |