Tissue-specific knockout of E-cadherin (Cdh1) in developing mouse gonads causes germ cells loss

in Reproduction

Correspondence should be addressed to R P Piprek; Email: rafal.piprek@uj.edu.pl
Restricted access

The normal course of gonad development is critical for the sexual development and reproductive capacity of the individual. During development, an incipient bipotential gonad which consists of unorganized aggregate of cells, must differentiate into highly structured testis or ovary. Cell adhesion molecules (CAMs) are a group of proteins crucial for segregation and aggregation of different cell types to form different tissues. E-cadherin (Cdh1) is one of the CAMs expressed in the developing gonads. We used tissue-specific knockout of Cdh1 gene in OCT4+ germ cells and, separately, in SF1+ somatic cells of developing gonads. The knockout of E-cadherin in somatic cells caused decrease in the number of germ cells, while the knockout in the germ cells caused their almost complete loss. Thus, the presence of E-cadherin in both the germ and somatic cells is necessary for the survival of germ cells. Although the lack of E-cadherin did not impair cell proliferation, it enhanced apoptosis, which was a possible cause of germ cell loss. However, the somatic cells of the gonad differentiated normally into Sertoli cells in the testis cords, and into follicular cells in the ovaries. The testis and ovigerous cords maintained their integrity; they were covered by continuous basement membranes. The testicular interstitium with steroidogenic fetal Leydig cells did not show any noticeable changes. However, in the female gonads, because of the lack of germ cells, the ovarian follicles were absent. The sex determination and sexual differentiation of the gonad were not impaired. These results underscore an important role of E-cadherin in germ cell survival and gonad development.

Supplementary Materials

    • Suppl. Fig. S1. Cre recombinase expression under the control of Oct4 promoter and the expression of Cdh1 (E-cadherin). A,B. A high expression of cre gene under the control of Oct4 promoter was detected in SSEA1+ cells isolated from XY and XX gonads from E11.5 onwards, and the expression level decreased at 2 dpp. C,D. The Cdh1 expression was comparable in SSEA1+ cells isolated from XY and XX gonads at E10.5 and E11.5, however, the expression was residual lowered at E13.5, and absent from E16.5 onwards. E,F. The expression of germ cell markers (Oct4, Mvh) and gonadal somatic cell markers (Sox9, Foxl2) in SSEA1+ cells isolated from Oct4-cre+ Cdh1fl/fl XY and XX gonads at E16.5. SSEA1+ cells showed high expression of germ cell markers when compared to SSEA- cells that expressed somatic markers. This confirmed that isolated SSEA1+ cells were the germ cells.
    • Suppl. Fig. S2. Cre recombinase expression under the control of Sf1 promoter and the expression of Cdh1 (E-cadherin). A,B. The expression of cre transcript was detected from E11.5 onwards in the isolated Pcdh18+ gonadal cells. C,D. The Cdh1 expression was comparable in Pcdh18+ cells isolated from XY and XX gonads at E10.5, however, the expression decreased from E11.5, and was absent from E13.5 in XY gonads, and from E16.5 in XX gonads. E,F. The expression of germ cell markers (Oct4, Mvh) and gonadal somatic cell markers (Sox9, Foxl2) in Pcdh18+ cells isolated from Sf1-cre+ Cdh1fl/fl XY and XX gonads at E16.5. Pcdh18+ cells showed high expression of germ cell markers in comparison to Pcdh18- cells that expressed somatic markers. This confirmed that isolated Pcdh18+ cells were the gonadal somatic cells.
    • Suppl. Table S1. Number of specimens tested.
    • Suppl. Table S2. Primers used for genotyping and RT-qPCR.
    • Suppl. Table S3. The number of germ cells per cross section of the control and knockout testes at E18.5.
    • Suppl. Table S4. The number of germ cells per cross section of the control and knockout ovaries at E18.5.
    • Suppl. Table S5. PCNA positive cells per cross section of the control and knockout testes at E18.5.
    • Suppl. Table S6. PCNA positive cells per cross section of the control and knockout ovaries at E18.5.
    • Suppl. Table S7. Caspase3-positive cells per cross section of the control and knockout testes at E18.5.
    • Suppl. Table S8. Caspase3 positive cells per cross section of the control and knockout ovaries at E18.5.
    • Suppl. Table S9. Cyp17a1 positive cells per cross section of the control and knockout testes at E18.5.

 

     An official journal of

    Society for Reproduction and Fertility

 

Sept 2018 onwards Past Year Past 30 Days
Abstract Views 1327 1199 122
Full Text Views 135 103 6
PDF Downloads 60 53 6