Dynamics of extracellular matrix in ovarian follicles and corpora lutea of mice

Despite the mouse being an important laboratory species, little is known about changes in its extracellular matrix (ECM) during follicle and corpora lutea formation and regression. Follicle development was induced in mice (29 days of age/experimental day 0) by injections of pregnant mare’s serum gonadotrophin on days 0 and 1 and ovulation was induced by injection of human chorionic gonadotrophin on day 2. Ovaries were collected for immunohistochemistry (n=10 per group) on days 0, 2 and 5. Another group was mated and ovaries were examined on day 11 (n=7). Collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2 and perlecan were present in the follicular basal lamina of all developmental stages. Collagen type XVIII was only found in basal lamina of primordial, primary and some preantral follicles, whereas laminin α2 was only detected in some preantral and antral follicles. The focimatrix, a specialised matrix of the membrana granulosa, contained collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. In the corpora lutea, staining was restricted to capillary sub-endothelial basal laminas containing collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. Laminins α4 and α5 were not immunolocalised to any structure in the mouse ovary. The ECM composition of the mouse ovary has similarities to, but also major differences from, other species with respect to nidogens 1 and 2 and perlecan

Seminiferous cord formation and germ-cell programming: epigenetic transgenerational actions of endocrine disruptors

The molecular and cellular control of embryonic testis development was investigated through an analysis of the embryonic testis transcriptome to identify potential regulatory factors for male sex determination and testis morphogenesis. One critical factor identified is neurotropin 3 (NT3). At the onset of male sex determination, Sertoli cells initiate differentiation and express NT3 to act as a chemotactic factor for mesonephros cells to migrate and associate with Sertoli-germ cell aggregates to promote cord formation. Promoter analysis suggests that NT3 may be an initial downstream gene to SRY and helps promote testis morphogenesis. Endocrine disruptors were used to potentially interfere with embryonic testis development and further investigate this biological process. The estrogenic pesticide methoxychlor and antiandrogenic fungicide vinclozolin were used. Previous studies have shown that methoxychlor and vinclozolin both interfere with embryonic testis cord formation and cause increased spermatogenic cell apoptosis in the adult testis. Interestingly, transient in vivo exposure to endocrine disruptors at the time of male sex determination caused a transgenerational phenotype (F1-F4) of spermatogenic cell apoptosis and subfertility. This apparent epigenetic mechanism involves altered DNA methylation and permanent re-programming of the male germ-line. A series of genes with altered DNA methylation and imprinting are being identified. Observations reviewed demonstrate that a transient embryonic in utero exposure to an endocrine disruptor influences the embryonic testis transcriptome and through epigenetic effects (e.g., DNA methylation) results in abnormal germ-cell differentiation that subsequently influences adult spermatogenic capacity and male fertility, and that this phenotype is transgenerational through the germ-line. The novel observations of transgenerational epigenetic endocrine disruptor actions on male reproduction critically impact the potential hazards of these compounds as environmental toxins. The literature reviewed provides insight into the molecular and cellular control of embryonic testis development, male sex determination, and the programming of the male germ-line

Extracellular matrix and its role in spermatogenesis

In adult mammalian testes, such as rats, Sertoli and germ cells at different stages of their development in the seminiferous epithelium are in close contact with the basement membrane, a modified form of extracellular matrix (ECM). In essence, Sertoli and germ cells in particular spermatogonia are “resting” on the basement membrane at different stages of the seminiferous epithelial cycle, relying on its structural and hormonal supports. Thus, it is not entirely unexpected that ECM plays a significant role in regulating spermatogenesis, particularly spermatogonia and Sertoli cells, and the blood-testis barrier (BTB) constituted by Sertoli cells since these cells are in physical contact with the basement membrane. Additionally, the basement membrane is also in close contact with the underlying collagen network and the myoid cell layers, which together with the lymphatic network, constitute the tunica propria. The seminiferous epithelium and the tunica propria, in turn, constitute the seminiferous tubule, which is the functional unit that produces spermatozoa via its interaction with Leydig cells in the interstitium. In short, the basement membrane and the underlying collagen network that create the acellular zone of the tunica propria may even facilitate cross-talk between the seminiferous epithelium, the myoid cells and cells in the interstitium. Recent studies in the field have illustrated the crucial role of ECM in supporting Sertoli and germ cell function in the seminiferous epithelium, including the BTB dynamics. In this chapter, we summarize some of the latest findings in the field regarding the functional role of ECM in spermatogenesis using the adult rat testis as a model. We also high light specific areas of research that deserve attention for investigators in the field

Extracellular Matrix and Its Role in Spermatogenesis

In adult mammalian testes, such as rats, Sertoli and germ cells at different stages of their development in the seminiferous epithelium are in close contact with the basement membrane, a modified form of extracellular matrix (ECM). In essence, Sertoli and germ cells in particular spermatogonia are “resting” on the basement membrane at different stages of the seminiferous epithelial cycle, relying on its structural and hormonal supports. Thus, it is not entirely unexpected that ECM plays a significant role in regulating spermatogenesis, particularly spermatogonia and Sertoli cells, and the blood-testis barrier (BTB) constituted by Sertoli cells since these cells are in physical contact with the basement membrane. Additionally, the basement membrane is also in close contact with the underlying collagen network and the myoid cell layers, which together with the lymphatic network, constitute the tunica propria. The seminiferous epithelium and the tunica propria, in turn, constitute the seminiferous tubule, which is the functional unit that produces spermatozoa via its interaction with Leydig cells in the interstitium. In short, the basement membrane and the underlying collagen network that create the acellular zone of the tunica propria may even facilitate cross-talk between the seminiferous epithelium, the myoid cells and cells in the interstitium. Recent studies in the field have illustrated the crucial role of ECM in supporting Sertoli and germ cell function in the seminiferous epithelium, including the BTB dynamics. In this chapter, we summarize some of the latest findings in the field regarding the functional role of ECM in spermatogenesis using the adult rat testis as a model. We also highlight specific areas of research that deserve attention for investigators in the field