Transcriptional profiling validates involvement of extracellular matrix and proteinases genes in mouse gonad development

Abstract

Extracellular matrix (ECM) plays an important scaffolding role in the establishment of organs structure during development. A great number of ECM components and enzymes (proteinases) regulating formation/degradation of ECM during organ remodeling have been identified. In order to study the role of ECM in the mouse gonad development, especially during sexual differentiation of the gonads when the structure of the testis and ovary becomes established, we performed a global analysis of transcriptome in three main cell types of developing gonad (supporting, interstitial/stromal and germ cells) using transgenic mice, cell sorting and microarray. The genes coding for ECM components were mostly expressed in two gonadal cell lines: supporting and interstitial/stromal cells. These two cell lines differed in the expression pattern of ECM components, which suggests that ECM components might be crucial for differentiation of gonad compartments (for example testis cords vs. interstitium in XY gonads). Collagens and proteoglycans coding genes were mainly expressed in the interstitium/stromal cells, while non-collagen glycoproteins and matricellular coding genes were expressed in both cell lines. We also analyzed the expression of genes encoding ECM enzymes that are secreted to the ECM where they remodel the scaffolding of developing organs. We found that the ECM enzyme genes were also mostly expressed in supporting and interstitial/stromal cells. In contrast to the somatic cells, the germ cells expressed only limited number of ECM components and enzymes. This suggests that the germ line cells do not participate, or play only a minor role, in the sculpting of the gonad structure via ECM synthesis and remodeling. Importantly, the supporting cells showed the sex-specific pattern of expression of ECM components. However, the pattern of expression of most ECM enzymes in the somatic and germ cells is independent on the sex of the gonad. Further studies are required to elucidate the exact roles of identified genes in sexual differentiation of the gonads.

Dissecting the initiation of female meiosis in the mouse at single-cell resolution

“…Pregranulosa cells or their precursors (supporting somatic cells) were identified within three distinct clusters on the basis of the elevated expression of Wnt4 and Wnt6 (Supplementary Figure 1d) (Jameson et al, 2012, McLaren, 2000). We also identified 5 ovarian somatic cells with their classic makers including mesothelial cells ( Lhx9 and Upk3b ) (Kanamori-Katayama et al, 2011, Mazaud et al, 2002), interstitial cells ( Col1a2 and Bgn ) (Piprek et al, 2018), endothelial cells ( Pecam1 and Kdr ) (Brennan et al, 2002, Jeays-Ward et al, 2003) and two contaminative somatic cell populations from blood, immune cells ( Cd52 and Car2 ) and erythroid cells ( Alas2 and Alad ) (Harigae et al, 2003, Lummertz da Rocha et al, 2018) (Supplementary Figure 1d). Thereby providing evidence of the heterogeneity of all somatic cell populations during the time frame analysed.…”

Action of the Metalloproteinases in Gonadal Remodeling during Sex Reversal in the Sequential Hermaphroditism of the Teleostei Fish Synbranchus marmoratus (Synbranchiformes: Synbranchidae)

“…The detection of the activity of enzyme 3β-hydroxysteroid dehydrogenase (3β-HSD) is used as a universal marker for Leydig cells [ 56 ]. The 3β-HSD enzyme is involved in several steps of the biosynthesis of steroid hormones, being responsible for the conversion of pregnenolone to progesterone; 17α-hydroxypregnenolone to 17α-hydroxyprogesterone; dehydroepiandrosterone to androstenedione; and androstenediol to testosterone [ 26 , 57 ]. Thus, it can be assumed that, at the time of its detection, plasma levels of testosterone should be high or at least present.…”

“…In mammals, studies show that some of the major MMPs are involved in remodeling processes of the male germinal epithelium [ 24 ] and in the rupture of the ovarian follicle [ 25 ]. A recent study also shows the expression of proteinase genes and proteolytic enzymes in gonad development of the mouse [ 26 ]. However, in aspects regarding the reproductive biology in teleosts, there are few studies that relate gonadal remodeling to the action of the MMPs and none regarding their role during sex reversal.…”

Activin A-derived human embryonic stem cells show increased competence to differentiate into primordial germ cell-like cells

“…The obtained hPGCLCs not only expressed high levels of early PGC markers, but also expressed markers of migratory PGCs, such as ITGA6 , CXCR4 , CDH4 , and LAMA4 21,22,60 . This may have been supported by the upregulation in the EB‐somatic cells of ECM ( COL3A1 /collagen type III alpha 1 chain) and ECM‐related ( SPARC /secreted protein acidic and cysteine rich) genes that are also expressed in gonadal supporting cells 71 . In this regard, it will be interesting to further investigate the maturation potential of the obtained hPGCLCs through long‐term coculture with mouse gonadal somatic cells 25 to facilitate meiotic entry and eventually generate functional gametes.…”