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The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle.

Erickson GF, Shimasaki S - Reprod. Biol. Endocrinol. (2003)

Bottom Line: We have found that: i) all of the mRNAs are expressed in a cell-specific manner in the major classes of ovary cell types (oocyte, granulosa, theca interstitial, theca externa, corpora lutea, secondary interstitial, vascular and ovary surface epithelium); and ii) most undergo dynamic changes during follicular and corpora luteal morphogenesis and histogenesis.The general principle to emerge from these studies is that the developmental programs of folliculogenesis (recruitment, selection, atresia), ovulation, and luteogenesis (luteinization, luteolysis) are accompanied by rather dramatic spatial and temporal changes in the expression patterns of these BMP genes.These results lead us to hypothesize previously unanticipated roles for the BMP family in determining fundamental developmental events that ensure the proper timing and developmental events required for the generation of the estrous cycle.

View Article: PubMed Central - HTML - PubMed

Affiliation: University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0674, USA. gerickson@ucsd.edu

ABSTRACT
In the mammalian ovary, great interest in the expression and function of the bone morphogenetic protein (BMP) family has been recently generated from evidence of their critical role in determining folliculogenesis and female fertility. Despite extensive work, there is a need to understand the cellular sites of expression of these important regulatory molecules, and how their gene expression changes within the basic ovary cell types through the cycle. Here we have performed a detailed in situ hybridization analysis of the spatial and temporal expression patterns of the BMP ligands (BMP-2, -3, -3b, -4, -6, -7, -15), receptors (BMPR-IA, -IB, -II), and BMP antagonist, follistatin, in rat ovaries over the normal estrous cycle. We have found that: i) all of the mRNAs are expressed in a cell-specific manner in the major classes of ovary cell types (oocyte, granulosa, theca interstitial, theca externa, corpora lutea, secondary interstitial, vascular and ovary surface epithelium); and ii) most undergo dynamic changes during follicular and corpora luteal morphogenesis and histogenesis. The general principle to emerge from these studies is that the developmental programs of folliculogenesis (recruitment, selection, atresia), ovulation, and luteogenesis (luteinization, luteolysis) are accompanied by rather dramatic spatial and temporal changes in the expression patterns of these BMP genes. These results lead us to hypothesize previously unanticipated roles for the BMP family in determining fundamental developmental events that ensure the proper timing and developmental events required for the generation of the estrous cycle.

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In situ hybridization of BMP-2 mRNA in the ovaries of adult cycling rats. Brightfield (A, D, F, H, J, L, N) and Darkfield (B, C, E, G, I, K, M). A, B: Sections from a DII 1100 h ovary (4X); new corpus luteum (CL-I); atretic follicle (AF); secondary interstitial cells (SIC); healthy Graafian follicle (GF); C: Negative control hybridized with sense BMP-2 cRNA probe; D, E: Secondary follicle with 3–5 layers of GCs at DII 1100 h (20X). Note positive membrana GC at periphery. F, G: Dominant follicle at DII 1100 h (10X); Positive membrana GC (MGC); Negative oocyte (O), cumulus GC (CC), periantral GC (PA) and theca (T); H, I: Atretic follicle at P 1000 h showing strongly positive GC (10X); J, K: Negative newly formed CL-I at E 0200 h (4X); Positive GF and preantral follicle (arrow); L, M: New CL-I after initiation of luteolysis at DII 1100 h showing clusters of positive cells (20X); N: Higher magnification (40X) of CL-I in L showing positive endothelial and granulosa lutein cells (GLC).
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Figure 1: In situ hybridization of BMP-2 mRNA in the ovaries of adult cycling rats. Brightfield (A, D, F, H, J, L, N) and Darkfield (B, C, E, G, I, K, M). A, B: Sections from a DII 1100 h ovary (4X); new corpus luteum (CL-I); atretic follicle (AF); secondary interstitial cells (SIC); healthy Graafian follicle (GF); C: Negative control hybridized with sense BMP-2 cRNA probe; D, E: Secondary follicle with 3–5 layers of GCs at DII 1100 h (20X). Note positive membrana GC at periphery. F, G: Dominant follicle at DII 1100 h (10X); Positive membrana GC (MGC); Negative oocyte (O), cumulus GC (CC), periantral GC (PA) and theca (T); H, I: Atretic follicle at P 1000 h showing strongly positive GC (10X); J, K: Negative newly formed CL-I at E 0200 h (4X); Positive GF and preantral follicle (arrow); L, M: New CL-I after initiation of luteolysis at DII 1100 h showing clusters of positive cells (20X); N: Higher magnification (40X) of CL-I in L showing positive endothelial and granulosa lutein cells (GLC).

Mentions: After an autoradiography and counterstaining, the sections were analyzed microscopically. The in situ experiments were performed two times for each BMP ligand and receptor. The intensity of the hybridization signals were determined as previously reported [26-28]. A subjective comparison of eight sections from each ovary from six animals hybridized with the same concentrations of sense and antisense cRNAs and exposed for the same time (4 weeks) was made. The hybridization signal was estimated on a scale of 1+ to 4+ as originally described by Meunier [29]: +, silver grains sparse, but positive hybridization; ++, silver grains are numerous but do not cover the cell type in question; +++, silver grains are very numerous and begin to merge in some places; ++++, silver grains are very dense and form a near uniform mass above the cell type in question; +/-, there was heterogeneity in the hybridization signal, e.g., some of the histological units contained a signal, others did not; –, no detectable hybridization signal. The slides were analyzed simultaneously by two investigators and the results confirmed independently by another researcher. In each experiment, tissues hybridized with the sense probe were used as the negative control. In all cases, the sense probes showed no specific hybridization to the adult ovaries above background (see for example Fig. 1C).


The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle.

Erickson GF, Shimasaki S - Reprod. Biol. Endocrinol. (2003)

In situ hybridization of BMP-2 mRNA in the ovaries of adult cycling rats. Brightfield (A, D, F, H, J, L, N) and Darkfield (B, C, E, G, I, K, M). A, B: Sections from a DII 1100 h ovary (4X); new corpus luteum (CL-I); atretic follicle (AF); secondary interstitial cells (SIC); healthy Graafian follicle (GF); C: Negative control hybridized with sense BMP-2 cRNA probe; D, E: Secondary follicle with 3–5 layers of GCs at DII 1100 h (20X). Note positive membrana GC at periphery. F, G: Dominant follicle at DII 1100 h (10X); Positive membrana GC (MGC); Negative oocyte (O), cumulus GC (CC), periantral GC (PA) and theca (T); H, I: Atretic follicle at P 1000 h showing strongly positive GC (10X); J, K: Negative newly formed CL-I at E 0200 h (4X); Positive GF and preantral follicle (arrow); L, M: New CL-I after initiation of luteolysis at DII 1100 h showing clusters of positive cells (20X); N: Higher magnification (40X) of CL-I in L showing positive endothelial and granulosa lutein cells (GLC).
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Related In: Results  -  Collection

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Figure 1: In situ hybridization of BMP-2 mRNA in the ovaries of adult cycling rats. Brightfield (A, D, F, H, J, L, N) and Darkfield (B, C, E, G, I, K, M). A, B: Sections from a DII 1100 h ovary (4X); new corpus luteum (CL-I); atretic follicle (AF); secondary interstitial cells (SIC); healthy Graafian follicle (GF); C: Negative control hybridized with sense BMP-2 cRNA probe; D, E: Secondary follicle with 3–5 layers of GCs at DII 1100 h (20X). Note positive membrana GC at periphery. F, G: Dominant follicle at DII 1100 h (10X); Positive membrana GC (MGC); Negative oocyte (O), cumulus GC (CC), periantral GC (PA) and theca (T); H, I: Atretic follicle at P 1000 h showing strongly positive GC (10X); J, K: Negative newly formed CL-I at E 0200 h (4X); Positive GF and preantral follicle (arrow); L, M: New CL-I after initiation of luteolysis at DII 1100 h showing clusters of positive cells (20X); N: Higher magnification (40X) of CL-I in L showing positive endothelial and granulosa lutein cells (GLC).
Mentions: After an autoradiography and counterstaining, the sections were analyzed microscopically. The in situ experiments were performed two times for each BMP ligand and receptor. The intensity of the hybridization signals were determined as previously reported [26-28]. A subjective comparison of eight sections from each ovary from six animals hybridized with the same concentrations of sense and antisense cRNAs and exposed for the same time (4 weeks) was made. The hybridization signal was estimated on a scale of 1+ to 4+ as originally described by Meunier [29]: +, silver grains sparse, but positive hybridization; ++, silver grains are numerous but do not cover the cell type in question; +++, silver grains are very numerous and begin to merge in some places; ++++, silver grains are very dense and form a near uniform mass above the cell type in question; +/-, there was heterogeneity in the hybridization signal, e.g., some of the histological units contained a signal, others did not; –, no detectable hybridization signal. The slides were analyzed simultaneously by two investigators and the results confirmed independently by another researcher. In each experiment, tissues hybridized with the sense probe were used as the negative control. In all cases, the sense probes showed no specific hybridization to the adult ovaries above background (see for example Fig. 1C).

Bottom Line: We have found that: i) all of the mRNAs are expressed in a cell-specific manner in the major classes of ovary cell types (oocyte, granulosa, theca interstitial, theca externa, corpora lutea, secondary interstitial, vascular and ovary surface epithelium); and ii) most undergo dynamic changes during follicular and corpora luteal morphogenesis and histogenesis.The general principle to emerge from these studies is that the developmental programs of folliculogenesis (recruitment, selection, atresia), ovulation, and luteogenesis (luteinization, luteolysis) are accompanied by rather dramatic spatial and temporal changes in the expression patterns of these BMP genes.These results lead us to hypothesize previously unanticipated roles for the BMP family in determining fundamental developmental events that ensure the proper timing and developmental events required for the generation of the estrous cycle.

View Article: PubMed Central - HTML - PubMed

Affiliation: University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0674, USA. gerickson@ucsd.edu

ABSTRACT
In the mammalian ovary, great interest in the expression and function of the bone morphogenetic protein (BMP) family has been recently generated from evidence of their critical role in determining folliculogenesis and female fertility. Despite extensive work, there is a need to understand the cellular sites of expression of these important regulatory molecules, and how their gene expression changes within the basic ovary cell types through the cycle. Here we have performed a detailed in situ hybridization analysis of the spatial and temporal expression patterns of the BMP ligands (BMP-2, -3, -3b, -4, -6, -7, -15), receptors (BMPR-IA, -IB, -II), and BMP antagonist, follistatin, in rat ovaries over the normal estrous cycle. We have found that: i) all of the mRNAs are expressed in a cell-specific manner in the major classes of ovary cell types (oocyte, granulosa, theca interstitial, theca externa, corpora lutea, secondary interstitial, vascular and ovary surface epithelium); and ii) most undergo dynamic changes during follicular and corpora luteal morphogenesis and histogenesis. The general principle to emerge from these studies is that the developmental programs of folliculogenesis (recruitment, selection, atresia), ovulation, and luteogenesis (luteinization, luteolysis) are accompanied by rather dramatic spatial and temporal changes in the expression patterns of these BMP genes. These results lead us to hypothesize previously unanticipated roles for the BMP family in determining fundamental developmental events that ensure the proper timing and developmental events required for the generation of the estrous cycle.

Show MeSH
Related in: MedlinePlus