The Experts below are selected from a list of 204 Experts worldwide ranked by ideXlab platform
Raymond J. Rodgers - One of the best experts on this subject based on the ideXlab platform.
-
Genetic relationships between early menopause and the behaviour of Theca Interna during follicular atresia.
Human reproduction (Oxford England), 2020Co-Authors: Raymond J. Rodgers, Joop S.e. LavenAbstract:Genetic variants are known to contribute to about 50% of the heritability of the age of menopause and recent studies suggest that genes associated with genome maintenance are involved. The idea that increased rates of follicular atresia could lead to depletion of the primoridial follicle reserve and early menopause has also been canvassed, but there is no direct evidence of this. In studies of the transcriptomics of follicular atresia, it was found that in the Theca Interna, the largest group of genes are in fact down-regulated and associated with 'cell cycle and DNA replication', in contrast with the up-regulation of apoptosis-associated genes which occurs in granulosa cells. Many of the genes down-regulated in the Theca Interna are the same as or related to the genes in loci associated with early menopause. From these findings, we suggest that early menopause could be due to increased rates of follicular atresia initiated from the Theca Interna.
-
Transcriptome analyses of ovarian stroma; tunica albuginea, interstitium and Theca Interna.
Reproduction (Cambridge England), 2019Co-Authors: Katja Hummitzsch, Nicholas Hatzirodos, Raymond J. Rodgers, Anne M. Macpherson, Jeffrey Schwartz, Helen F. Irving-rodgersAbstract:The ovary has specialised stromal compartments, including the tunica albuginea, interstitial stroma and Theca Interna, which develops concurrently with the follicular antrum. To characterise the molecular determinants of these compartments, stroma adjacent to preantral follicles (pre-Theca), interstitium and tunica albuginea were laser microdissected (n = 4 per group) and Theca Interna was dissected from bovine antral follicles (n = 6). RNA microarray analysis showed minimal differences between interstitial stroma and pre-Theca, and these were combined for some analyses and referred to as stroma. Genes significantly upregulated in Theca Interna compared to stroma included INSL3, LHCGR, HSD3B1, CYP17A1, ALDH1A1, OGN, POSTN and ASPN. Quantitative RT-PCR showed significantly greater expression of OGN and LGALS1 in interstitial stroma and Theca Interna versus tunica and greater expression of ACD in tunica compared to Theca Interna. PLN was significantly higher in interstitial stroma compared to tunica and Theca. Ingenuity pathway, network and upstream regulator analyses were undertaken. Cell survival was also upregulated in Theca Interna. The tunica albuginea was associated with GPCR and cAMP signalling, suggesting tunica contractility. It was also associated with TGF-β signalling and increased fibrous matrix. Western immunoblotting was positive for OGN, LGALS1, ALDH1A1, ACD and PLN with PLN and OGN highly expressed in tunica and interstitial stroma (each n = 6), but not in Theca Interna from antral follicles (n = 24). Immunohistochemistry localised LGALS1 and POSTN to extracellular matrix and PLN to smooth muscle cells. These results have identified novel differences between the ovarian stromal compartments.
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts - Fig 2
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:Unsupervised PCA of granulosa cell (A) and Theca Interna (B) arrays. The graphs are scatter plots of the values for the first (X) and second (Y) principal components based on the correlation matrix of the total normalised array intensity data. The numbering of each sample enables the samples in the figure to be identified in S1 and S2 Figs. Abbreviations are GC (granulosa cell), TH (Theca Interna), CC (cultured under control conditions), CF (cultured and FSH treated), CL (cultured and LH treated) and V (in vivo freshly isolated cells).
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:In vitro culture of ovarian granulosa cells and Theca cells has been very important for our understanding of their function and regulation. One of the most eagerly sought attributes of cell culture is the use of chemically-defined conditions. However, even under such in vitro conditions cell behaviour could differ from the in vivo situation because of differences in oxygen tension, nutrients, adhesion matrix and other factors. To examine this further we compared the transcriptomes of both granulosa cells and cells from the Theca Interna that were cultured in what are arguably the best in vitro conditions for maintaining the ‘follicular’ phenotypes of both tissue types, as displayed by their respective freshly-isolated counterparts. The array data analysed are from recently published data and use the same sizes of bovine follicles (small antral 3–6 mm) and the same Affymetrix arrays. We conducted analysis using Partek, Ingenuity Pathway Analysis and GOEAST. Principal Component Analysis (PCA) and hierarchical clustering clearly separated the in vivo from the in vitro groups for both cells types and transcriptomes were more homogeneous upon culture. In both cell cultures behaviours associated with cell adhesion, migration and interaction with matrix or substrate were more abundant. However, the pathways involved generally differed between the two cell types. With the Thecal cultures a gene expression signature of an immune response was more abundant, probably by leukocytes amongst the cells cultured from the Theca Interna. These results indicate differences between in vivo and in vitro that should be considered when interpreting in vitro data.
-
Transcriptome Profiling of the Theca Interna from Bovine Ovarian Follicles
2016Co-Authors: During Atresia, Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna is a specialized stromal layer that envelops each growing ovarian follicle. It contains capillaries, fibroblasts, immune cells and the steroidogenic cells that synthesize androgens for conversion to estradiol by the neighboring granulosa cells. During reproductive life only a small number of follicles will grow to a sufficient size to ovulate, whereas the majority of follicles will undergo regression/atresia and phagocytosis by macrophages. To identify genes which are differentially regulated in the Theca Interna during follicular atresia, we undertook transcriptome profiling of the Theca Interna from healthy (n = 10) and antral atretic (n = 5) bovine follicles at early antral stages (,5 mm). Principal Component Analyses and hierarchical classification of the signal intensity plots for the arrays showed primary clustering into two groups, healthy and atretic. A total of 543 probe sets were differentially expressed between the atretic and healthy Theca Interna. Further analyses of these genes by Ingenuity Pathway Analysis and Gene Ontology Enrichment Analysis Toolkit software found most of the genes being expressed were related to cytokines, hormones and receptors as well as the cell cycle and DNA replication. Cell cycle genes which encode components of the replicating chromosome complex and mitotic spindle were down-regulated in atretic Theca Interna, whereas stress response and inflammation-related genes such as TP53, IKBKB and TGFB1 were up-regulated. In addition to cell cycle regulators, upstream regulators that were predicted to be inhibited included Retinoblastoma 1, E2 transcription factor 1, and hepatocyte growth factor. Our study suggests that during antra
Nicholas Hatzirodos - One of the best experts on this subject based on the ideXlab platform.
-
Transcriptome analyses of ovarian stroma; tunica albuginea, interstitium and Theca Interna.
Reproduction (Cambridge England), 2019Co-Authors: Katja Hummitzsch, Nicholas Hatzirodos, Raymond J. Rodgers, Anne M. Macpherson, Jeffrey Schwartz, Helen F. Irving-rodgersAbstract:The ovary has specialised stromal compartments, including the tunica albuginea, interstitial stroma and Theca Interna, which develops concurrently with the follicular antrum. To characterise the molecular determinants of these compartments, stroma adjacent to preantral follicles (pre-Theca), interstitium and tunica albuginea were laser microdissected (n = 4 per group) and Theca Interna was dissected from bovine antral follicles (n = 6). RNA microarray analysis showed minimal differences between interstitial stroma and pre-Theca, and these were combined for some analyses and referred to as stroma. Genes significantly upregulated in Theca Interna compared to stroma included INSL3, LHCGR, HSD3B1, CYP17A1, ALDH1A1, OGN, POSTN and ASPN. Quantitative RT-PCR showed significantly greater expression of OGN and LGALS1 in interstitial stroma and Theca Interna versus tunica and greater expression of ACD in tunica compared to Theca Interna. PLN was significantly higher in interstitial stroma compared to tunica and Theca. Ingenuity pathway, network and upstream regulator analyses were undertaken. Cell survival was also upregulated in Theca Interna. The tunica albuginea was associated with GPCR and cAMP signalling, suggesting tunica contractility. It was also associated with TGF-β signalling and increased fibrous matrix. Western immunoblotting was positive for OGN, LGALS1, ALDH1A1, ACD and PLN with PLN and OGN highly expressed in tunica and interstitial stroma (each n = 6), but not in Theca Interna from antral follicles (n = 24). Immunohistochemistry localised LGALS1 and POSTN to extracellular matrix and PLN to smooth muscle cells. These results have identified novel differences between the ovarian stromal compartments.
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts - Fig 2
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:Unsupervised PCA of granulosa cell (A) and Theca Interna (B) arrays. The graphs are scatter plots of the values for the first (X) and second (Y) principal components based on the correlation matrix of the total normalised array intensity data. The numbering of each sample enables the samples in the figure to be identified in S1 and S2 Figs. Abbreviations are GC (granulosa cell), TH (Theca Interna), CC (cultured under control conditions), CF (cultured and FSH treated), CL (cultured and LH treated) and V (in vivo freshly isolated cells).
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:In vitro culture of ovarian granulosa cells and Theca cells has been very important for our understanding of their function and regulation. One of the most eagerly sought attributes of cell culture is the use of chemically-defined conditions. However, even under such in vitro conditions cell behaviour could differ from the in vivo situation because of differences in oxygen tension, nutrients, adhesion matrix and other factors. To examine this further we compared the transcriptomes of both granulosa cells and cells from the Theca Interna that were cultured in what are arguably the best in vitro conditions for maintaining the ‘follicular’ phenotypes of both tissue types, as displayed by their respective freshly-isolated counterparts. The array data analysed are from recently published data and use the same sizes of bovine follicles (small antral 3–6 mm) and the same Affymetrix arrays. We conducted analysis using Partek, Ingenuity Pathway Analysis and GOEAST. Principal Component Analysis (PCA) and hierarchical clustering clearly separated the in vivo from the in vitro groups for both cells types and transcriptomes were more homogeneous upon culture. In both cell cultures behaviours associated with cell adhesion, migration and interaction with matrix or substrate were more abundant. However, the pathways involved generally differed between the two cell types. With the Thecal cultures a gene expression signature of an immune response was more abundant, probably by leukocytes amongst the cells cultured from the Theca Interna. These results indicate differences between in vivo and in vitro that should be considered when interpreting in vitro data.
-
Transcriptome Profiling of the Theca Interna from Bovine Ovarian Follicles
2016Co-Authors: During Atresia, Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna is a specialized stromal layer that envelops each growing ovarian follicle. It contains capillaries, fibroblasts, immune cells and the steroidogenic cells that synthesize androgens for conversion to estradiol by the neighboring granulosa cells. During reproductive life only a small number of follicles will grow to a sufficient size to ovulate, whereas the majority of follicles will undergo regression/atresia and phagocytosis by macrophages. To identify genes which are differentially regulated in the Theca Interna during follicular atresia, we undertook transcriptome profiling of the Theca Interna from healthy (n = 10) and antral atretic (n = 5) bovine follicles at early antral stages (,5 mm). Principal Component Analyses and hierarchical classification of the signal intensity plots for the arrays showed primary clustering into two groups, healthy and atretic. A total of 543 probe sets were differentially expressed between the atretic and healthy Theca Interna. Further analyses of these genes by Ingenuity Pathway Analysis and Gene Ontology Enrichment Analysis Toolkit software found most of the genes being expressed were related to cytokines, hormones and receptors as well as the cell cycle and DNA replication. Cell cycle genes which encode components of the replicating chromosome complex and mitotic spindle were down-regulated in atretic Theca Interna, whereas stress response and inflammation-related genes such as TP53, IKBKB and TGFB1 were up-regulated. In addition to cell cycle regulators, upstream regulators that were predicted to be inhibited included Retinoblastoma 1, E2 transcription factor 1, and hepatocyte growth factor. Our study suggests that during antra
-
Transcriptome Profiling of the Theca Interna in Transition from Small to Large Antral Ovarian Follicles
2016Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna layer of the ovarian follicle forms during the antral stage of follicle development and lies adjacent to and directly outside the follicular basal lamina. It supplies androgens and communicates with the granulosa cells and the oocyte by extracellular signaling. To better understand developmental changes in the Theca Interna, we undertook transcriptome profiling of the Theca Interna from small (3–5 mm, n = 10) and large (9–12 mm, n = 5) healthy antral bovine follicles, representing a calculated.7-fold increase in the amount of Thecal tissue. Principal Component Analysis and hierarchical classification of the signal intensity plots for the arrays showed no clustering of the Theca Interna samples into groups depending on follicle size or subcategories of small follicles. From the over 23,000 probe sets analysed, only 76 were differentially expressed between large and small healthy follicles. Some of the differentially expressed genes were associated with processes such as myoblast differentiation, protein ubiquitination, nitric oxide and transforming growth factor b signaling. The most significant pathway affected from our analyses was found to be Wnt signaling, which was suppressed in large follicles via down-regulation of WNT2B and up-regulation of the inhibitor FRZB. These changes in the transcriptional profile could have been due to changes in cellular function or alternatively since the Theca Interna is composed of a number of different cell types it could have been due to any systematic change in the volume density of any particular cell type. However, our study suggests that the transcriptional profile of the Theca Interna is relatively stabl
Helen F. Irving-rodgers - One of the best experts on this subject based on the ideXlab platform.
-
Transcriptome analyses of ovarian stroma; tunica albuginea, interstitium and Theca Interna.
Reproduction (Cambridge England), 2019Co-Authors: Katja Hummitzsch, Nicholas Hatzirodos, Raymond J. Rodgers, Anne M. Macpherson, Jeffrey Schwartz, Helen F. Irving-rodgersAbstract:The ovary has specialised stromal compartments, including the tunica albuginea, interstitial stroma and Theca Interna, which develops concurrently with the follicular antrum. To characterise the molecular determinants of these compartments, stroma adjacent to preantral follicles (pre-Theca), interstitium and tunica albuginea were laser microdissected (n = 4 per group) and Theca Interna was dissected from bovine antral follicles (n = 6). RNA microarray analysis showed minimal differences between interstitial stroma and pre-Theca, and these were combined for some analyses and referred to as stroma. Genes significantly upregulated in Theca Interna compared to stroma included INSL3, LHCGR, HSD3B1, CYP17A1, ALDH1A1, OGN, POSTN and ASPN. Quantitative RT-PCR showed significantly greater expression of OGN and LGALS1 in interstitial stroma and Theca Interna versus tunica and greater expression of ACD in tunica compared to Theca Interna. PLN was significantly higher in interstitial stroma compared to tunica and Theca. Ingenuity pathway, network and upstream regulator analyses were undertaken. Cell survival was also upregulated in Theca Interna. The tunica albuginea was associated with GPCR and cAMP signalling, suggesting tunica contractility. It was also associated with TGF-β signalling and increased fibrous matrix. Western immunoblotting was positive for OGN, LGALS1, ALDH1A1, ACD and PLN with PLN and OGN highly expressed in tunica and interstitial stroma (each n = 6), but not in Theca Interna from antral follicles (n = 24). Immunohistochemistry localised LGALS1 and POSTN to extracellular matrix and PLN to smooth muscle cells. These results have identified novel differences between the ovarian stromal compartments.
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts - Fig 2
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:Unsupervised PCA of granulosa cell (A) and Theca Interna (B) arrays. The graphs are scatter plots of the values for the first (X) and second (Y) principal components based on the correlation matrix of the total normalised array intensity data. The numbering of each sample enables the samples in the figure to be identified in S1 and S2 Figs. Abbreviations are GC (granulosa cell), TH (Theca Interna), CC (cultured under control conditions), CF (cultured and FSH treated), CL (cultured and LH treated) and V (in vivo freshly isolated cells).
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:In vitro culture of ovarian granulosa cells and Theca cells has been very important for our understanding of their function and regulation. One of the most eagerly sought attributes of cell culture is the use of chemically-defined conditions. However, even under such in vitro conditions cell behaviour could differ from the in vivo situation because of differences in oxygen tension, nutrients, adhesion matrix and other factors. To examine this further we compared the transcriptomes of both granulosa cells and cells from the Theca Interna that were cultured in what are arguably the best in vitro conditions for maintaining the ‘follicular’ phenotypes of both tissue types, as displayed by their respective freshly-isolated counterparts. The array data analysed are from recently published data and use the same sizes of bovine follicles (small antral 3–6 mm) and the same Affymetrix arrays. We conducted analysis using Partek, Ingenuity Pathway Analysis and GOEAST. Principal Component Analysis (PCA) and hierarchical clustering clearly separated the in vivo from the in vitro groups for both cells types and transcriptomes were more homogeneous upon culture. In both cell cultures behaviours associated with cell adhesion, migration and interaction with matrix or substrate were more abundant. However, the pathways involved generally differed between the two cell types. With the Thecal cultures a gene expression signature of an immune response was more abundant, probably by leukocytes amongst the cells cultured from the Theca Interna. These results indicate differences between in vivo and in vitro that should be considered when interpreting in vitro data.
-
Transcriptome Profiling of the Theca Interna from Bovine Ovarian Follicles
2016Co-Authors: During Atresia, Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna is a specialized stromal layer that envelops each growing ovarian follicle. It contains capillaries, fibroblasts, immune cells and the steroidogenic cells that synthesize androgens for conversion to estradiol by the neighboring granulosa cells. During reproductive life only a small number of follicles will grow to a sufficient size to ovulate, whereas the majority of follicles will undergo regression/atresia and phagocytosis by macrophages. To identify genes which are differentially regulated in the Theca Interna during follicular atresia, we undertook transcriptome profiling of the Theca Interna from healthy (n = 10) and antral atretic (n = 5) bovine follicles at early antral stages (,5 mm). Principal Component Analyses and hierarchical classification of the signal intensity plots for the arrays showed primary clustering into two groups, healthy and atretic. A total of 543 probe sets were differentially expressed between the atretic and healthy Theca Interna. Further analyses of these genes by Ingenuity Pathway Analysis and Gene Ontology Enrichment Analysis Toolkit software found most of the genes being expressed were related to cytokines, hormones and receptors as well as the cell cycle and DNA replication. Cell cycle genes which encode components of the replicating chromosome complex and mitotic spindle were down-regulated in atretic Theca Interna, whereas stress response and inflammation-related genes such as TP53, IKBKB and TGFB1 were up-regulated. In addition to cell cycle regulators, upstream regulators that were predicted to be inhibited included Retinoblastoma 1, E2 transcription factor 1, and hepatocyte growth factor. Our study suggests that during antra
-
Transcriptome Profiling of the Theca Interna in Transition from Small to Large Antral Ovarian Follicles
2016Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna layer of the ovarian follicle forms during the antral stage of follicle development and lies adjacent to and directly outside the follicular basal lamina. It supplies androgens and communicates with the granulosa cells and the oocyte by extracellular signaling. To better understand developmental changes in the Theca Interna, we undertook transcriptome profiling of the Theca Interna from small (3–5 mm, n = 10) and large (9–12 mm, n = 5) healthy antral bovine follicles, representing a calculated.7-fold increase in the amount of Thecal tissue. Principal Component Analysis and hierarchical classification of the signal intensity plots for the arrays showed no clustering of the Theca Interna samples into groups depending on follicle size or subcategories of small follicles. From the over 23,000 probe sets analysed, only 76 were differentially expressed between large and small healthy follicles. Some of the differentially expressed genes were associated with processes such as myoblast differentiation, protein ubiquitination, nitric oxide and transforming growth factor b signaling. The most significant pathway affected from our analyses was found to be Wnt signaling, which was suppressed in large follicles via down-regulation of WNT2B and up-regulation of the inhibitor FRZB. These changes in the transcriptional profile could have been due to changes in cellular function or alternatively since the Theca Interna is composed of a number of different cell types it could have been due to any systematic change in the volume density of any particular cell type. However, our study suggests that the transcriptional profile of the Theca Interna is relatively stabl
Katja Hummitzsch - One of the best experts on this subject based on the ideXlab platform.
-
Transcriptome analyses of ovarian stroma; tunica albuginea, interstitium and Theca Interna.
Reproduction (Cambridge England), 2019Co-Authors: Katja Hummitzsch, Nicholas Hatzirodos, Raymond J. Rodgers, Anne M. Macpherson, Jeffrey Schwartz, Helen F. Irving-rodgersAbstract:The ovary has specialised stromal compartments, including the tunica albuginea, interstitial stroma and Theca Interna, which develops concurrently with the follicular antrum. To characterise the molecular determinants of these compartments, stroma adjacent to preantral follicles (pre-Theca), interstitium and tunica albuginea were laser microdissected (n = 4 per group) and Theca Interna was dissected from bovine antral follicles (n = 6). RNA microarray analysis showed minimal differences between interstitial stroma and pre-Theca, and these were combined for some analyses and referred to as stroma. Genes significantly upregulated in Theca Interna compared to stroma included INSL3, LHCGR, HSD3B1, CYP17A1, ALDH1A1, OGN, POSTN and ASPN. Quantitative RT-PCR showed significantly greater expression of OGN and LGALS1 in interstitial stroma and Theca Interna versus tunica and greater expression of ACD in tunica compared to Theca Interna. PLN was significantly higher in interstitial stroma compared to tunica and Theca. Ingenuity pathway, network and upstream regulator analyses were undertaken. Cell survival was also upregulated in Theca Interna. The tunica albuginea was associated with GPCR and cAMP signalling, suggesting tunica contractility. It was also associated with TGF-β signalling and increased fibrous matrix. Western immunoblotting was positive for OGN, LGALS1, ALDH1A1, ACD and PLN with PLN and OGN highly expressed in tunica and interstitial stroma (each n = 6), but not in Theca Interna from antral follicles (n = 24). Immunohistochemistry localised LGALS1 and POSTN to extracellular matrix and PLN to smooth muscle cells. These results have identified novel differences between the ovarian stromal compartments.
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts - Fig 2
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:Unsupervised PCA of granulosa cell (A) and Theca Interna (B) arrays. The graphs are scatter plots of the values for the first (X) and second (Y) principal components based on the correlation matrix of the total normalised array intensity data. The numbering of each sample enables the samples in the figure to be identified in S1 and S2 Figs. Abbreviations are GC (granulosa cell), TH (Theca Interna), CC (cultured under control conditions), CF (cultured and FSH treated), CL (cultured and LH treated) and V (in vivo freshly isolated cells).
-
Transcriptomal profiling of bovine ovarian granulosa and Theca Interna cells in primary culture in comparison with their in vivo counterparts
2017Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Claire Glister, Philip G. Knight, Raymond J. RodgersAbstract:In vitro culture of ovarian granulosa cells and Theca cells has been very important for our understanding of their function and regulation. One of the most eagerly sought attributes of cell culture is the use of chemically-defined conditions. However, even under such in vitro conditions cell behaviour could differ from the in vivo situation because of differences in oxygen tension, nutrients, adhesion matrix and other factors. To examine this further we compared the transcriptomes of both granulosa cells and cells from the Theca Interna that were cultured in what are arguably the best in vitro conditions for maintaining the ‘follicular’ phenotypes of both tissue types, as displayed by their respective freshly-isolated counterparts. The array data analysed are from recently published data and use the same sizes of bovine follicles (small antral 3–6 mm) and the same Affymetrix arrays. We conducted analysis using Partek, Ingenuity Pathway Analysis and GOEAST. Principal Component Analysis (PCA) and hierarchical clustering clearly separated the in vivo from the in vitro groups for both cells types and transcriptomes were more homogeneous upon culture. In both cell cultures behaviours associated with cell adhesion, migration and interaction with matrix or substrate were more abundant. However, the pathways involved generally differed between the two cell types. With the Thecal cultures a gene expression signature of an immune response was more abundant, probably by leukocytes amongst the cells cultured from the Theca Interna. These results indicate differences between in vivo and in vitro that should be considered when interpreting in vitro data.
-
Transcriptome Profiling of the Theca Interna from Bovine Ovarian Follicles
2016Co-Authors: During Atresia, Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna is a specialized stromal layer that envelops each growing ovarian follicle. It contains capillaries, fibroblasts, immune cells and the steroidogenic cells that synthesize androgens for conversion to estradiol by the neighboring granulosa cells. During reproductive life only a small number of follicles will grow to a sufficient size to ovulate, whereas the majority of follicles will undergo regression/atresia and phagocytosis by macrophages. To identify genes which are differentially regulated in the Theca Interna during follicular atresia, we undertook transcriptome profiling of the Theca Interna from healthy (n = 10) and antral atretic (n = 5) bovine follicles at early antral stages (,5 mm). Principal Component Analyses and hierarchical classification of the signal intensity plots for the arrays showed primary clustering into two groups, healthy and atretic. A total of 543 probe sets were differentially expressed between the atretic and healthy Theca Interna. Further analyses of these genes by Ingenuity Pathway Analysis and Gene Ontology Enrichment Analysis Toolkit software found most of the genes being expressed were related to cytokines, hormones and receptors as well as the cell cycle and DNA replication. Cell cycle genes which encode components of the replicating chromosome complex and mitotic spindle were down-regulated in atretic Theca Interna, whereas stress response and inflammation-related genes such as TP53, IKBKB and TGFB1 were up-regulated. In addition to cell cycle regulators, upstream regulators that were predicted to be inhibited included Retinoblastoma 1, E2 transcription factor 1, and hepatocyte growth factor. Our study suggests that during antra
-
Transcriptome Profiling of the Theca Interna in Transition from Small to Large Antral Ovarian Follicles
2016Co-Authors: Nicholas Hatzirodos, Katja Hummitzsch, Helen F. Irving-rodgers, Raymond J. RodgersAbstract:The Theca Interna layer of the ovarian follicle forms during the antral stage of follicle development and lies adjacent to and directly outside the follicular basal lamina. It supplies androgens and communicates with the granulosa cells and the oocyte by extracellular signaling. To better understand developmental changes in the Theca Interna, we undertook transcriptome profiling of the Theca Interna from small (3–5 mm, n = 10) and large (9–12 mm, n = 5) healthy antral bovine follicles, representing a calculated.7-fold increase in the amount of Thecal tissue. Principal Component Analysis and hierarchical classification of the signal intensity plots for the arrays showed no clustering of the Theca Interna samples into groups depending on follicle size or subcategories of small follicles. From the over 23,000 probe sets analysed, only 76 were differentially expressed between large and small healthy follicles. Some of the differentially expressed genes were associated with processes such as myoblast differentiation, protein ubiquitination, nitric oxide and transforming growth factor b signaling. The most significant pathway affected from our analyses was found to be Wnt signaling, which was suppressed in large follicles via down-regulation of WNT2B and up-regulation of the inhibitor FRZB. These changes in the transcriptional profile could have been due to changes in cellular function or alternatively since the Theca Interna is composed of a number of different cell types it could have been due to any systematic change in the volume density of any particular cell type. However, our study suggests that the transcriptional profile of the Theca Interna is relatively stabl
Yukinori Yoshimura - One of the best experts on this subject based on the ideXlab platform.
-
Comparison of cadherin and integrin localization in bovine cystic and healthy follicles
Animal science journal = Nihon chikusan Gakkaiho, 2012Co-Authors: Tomomi Ohira, Yukinori Yoshimura, Chiaki Murayama, Takashi Shimizu, Naoki IsobeAbstract:As stage progresses in the cystic follicle, granulosa cells are lost. We hypothesized that the granulosa and Theca Interna layers are detached in association with weakened expression of cell adhesion molecules such as cadherin (cell-cell adhesion) and integrin (cell-extracellular matrix adhesion) in cystic follicles. To elucidate this hypothesis, we immunolocalized these molecules in the granulosa and Theca Interna and compared them between cystic and small healthy follicles. Sections were immunostained with cadherin and integrin β1 antibodies and their localizations were compared. Cadherin-positive reaction was seen in the cytoplasma of all granulosa cells. No increase in the frequency of cadherin-positive area in the granulosa layers and the intensity of cadherin immunoreaction in the Theca Interna was detected in cystic follicles compared with healthy ones. A dense immunoreaction product of integrin β1 was detected in the Theca Interna in both cystic and healthy follicles. Intensity of integrin β1 -immuno reaction in the granulosa layers and integrin β1 -positive area in the Theca Interna was significantly lower in the cystic follicle than in the healthy follicles. These results suggest that granulosa and Theca Interna cells are detached while maintaining the cell-cell adhesion, resulting in the consequent loss of these layers from the cystic follicle.
-
Deficient Proliferation and Apoptosis in the Granulosa and Theca Interna Cells of the Bovine Cystic Follicle
The Journal of reproduction and development, 2007Co-Authors: Naoki Isobe, Yukinori YoshimuraAbstract:The aim of the present study was to examine the frequencies of cell proliferation and death of granulosa and Theca Interna layers during development of cystic follicles in order to understand the mechanisms of cystic follicle formation. Paraffin sections of cystic follicles were immunostained with antibodies against proliferating cell nuclear antigen (PCNA) and cleaved caspase-3 in order to observe proliferating and apoptotic cells, respectively. The concentrations of estradiol-17β and progesterone in the follicular fluid of these follicles were measured by ELISA. The granulosa and Theca Interna layers contained both PCNA- and caspase-3-positive cells, although their numbers were limited. There was significant negative correlation between the estradiol-17β and progesterone concentrations in the follicular fluid. Regression analysis revealed no significant correlation, except for that between the PCNA-positive cells in the Theca Interna and the caspase-3-positive cells in the granulosa layer. These results indicate that the granulosa and Theca Interna cells of the cystic follicle show weak proliferative activity and low apoptotic frequency; this implies that the cystic follicle grows slowly and then maintains a static condition without degeneration, which leads to long-term persistence of the follicle.
-
Microvascular distribution and vascular endothelial growth factor expression in bovine cystic follicles
Domestic animal endocrinology, 2005Co-Authors: Naoki Isobe, M. Kitabayashi, Yukinori YoshimuraAbstract:The aim of this study was to examine the distribution of microvessels in the Theca and the expression of vascular endothelial growth factor (VEGF) in the Theca and granulosa of cystic follicles. Paraffin sections of cystic follicles were stained with Bandeiraea simplicifolia-I (BS-I) to visualize the endothelial cells of microvessels. The other sections were immunostained with anti-VEGF antibody. The mRNA expression of VEGF in the Theca Interna of cystic and healthy follicle was determined by RT-PCR. In the Theca Interna, cystic follicles with granulosa cells had significantly greater microvessel number density (the number of microvessels per given field) and area (area occupied by microvessels per given area) than healthy follicles in various sizes ( 9 mm). Loss of granulosa cells from cystic follicles resulted in a similar number density, but significantly smaller area of microvessels in the Theca Interna. There was no significant difference in the microvessel number density and area of the Theca externa between the types of follicle. VEGF protein was expressed in the granulosa and Theca Interna of healthy and cystic follicles. These results demonstrate that cystic follicles have a highly developed vasculature network in the Theca Interna, especially in cystic follicles containing granulosa cells. It is also suggested that VEGF is highly expressed in the cystic follicle as well as healthy follicle, which may be associated with advanced vasculature and the accumulation of follicular fluid in cystic follicles.
-
Distribution of Cytochrome P450-side Chain Cleavage in the Theca Interna Layers of Bovine Small Antral and Cystic Follicles
Reproduction in domestic animals = Zuchthygiene, 2003Co-Authors: Naoki Isobe, Toshihiko Nakao, Yukinori YoshimuraAbstract:Cystic follicle is anovulatory follicular structure that is caused by an endocrine imbalance. The activity of cytochrome P450-side chain cleavage (P450scc) is essential for the initiation of steroidogenesis in the follicle. The present study was designed to compare the frequency of cells containing P450scc between healthy and atretic small antral follicles, and among several types (I, II and III, classified based on the presence of granulosa layer) of cystic follicles. Paraffin sections of healthy (2-5 mm in diameter), atretic (2-5 mm) and cystic follicles (>25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine P450scc. The P450scc-positive cells were counted in four different regions of the follicles from the apical to the basal side. In small antral follicles and cystic follicles, P450scc-positive cells were localized in the Theca Interna layers but not granulosa layers. The P450scc-positive cell populations decreased in the late atretic follicles compared with the early and advanced atretic follicles at all the regions of follicle. Type III cystic follicles showed significantly lower frequencies of P450scc-positive cells than those in the types I and II cystic follicles. These results suggest that in both small and cystic follicles in cows, total loss of granulosa cells may be associated with the reduction of frequency of P450scc-positive cells in Theca Interna layer.
-
immunohistochemical localization of 3β hydroxysteroid dehydrogenase in the granulosa and Theca Interna layers of bovine cystic follicles
Journal of Reproduction and Development, 2003Co-Authors: Naoki Isobe, Toshihiko Nakao, Yukinori YoshimuraAbstract:The aim of the present study was to determine whether the alteration of population of cells containing 3β-hydroxysteroid dehydrogenase (3β-HSD) is responsible for the formation of cystic follicles. Paraffin sections of healthy (2 to 5 mm in diameter), atretic (2 to 5 mm) and cystic follicles (more than 25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine 3β-HSD. The 3β-HSD-positive cells were counted in 4 different regions of the follicles from the apical to the basal side. The frequencies of 3β-HSD-positive granulosa cells in cystic follicles were significantly higher than those in the healthy follicles (P<0.05), although the number of 3β-HSD-positive granulosa cells in the cystic follicle were fewer than half the cells (30 to 40%) and was much smaller than that in preovulatory follicles (Conley et al., 1995). The frequencies of 3β-HSD-positive cells were higher in the granulosa layer and lower in the Theca Interna layer of the cystic follicles than the atretic follicles. These results suggest that the differentiation of granulosa cells to express 3β-HSD might be insufficient in cystic follicles and accordingly they fail to ovulate. The differences of frequencies of 3β-HSD-positive cells in the granulosa and Theca Interna layers between cystic and atretic follicles may be one of the reasons why regression is delayed in cystic follicles.