Fig. 3: Interspecific blastocyst complementation in germline generates functional mouse spermatids in rats. | Nature Communications

Fig. 3: Interspecific blastocyst complementation in germline generates functional mouse spermatids in rats.

From: Blastocyst complementation using Prdm14-deficient rats enables efficient germline transmission and generation of functional mouse spermatids in rats

Fig. 3

a Schematics showing injection of mouse ESCs or iPSCs into Prdm14-H2BVenus positive rat blastocysts. b Wholemount immunostaining of the posterior part of Prdm14mut/HV chimeric embryo with mouse ESCs at 6 days after embryo transfer (ET: embryonic day (E) 9ā€“9.5). Dashed line shows cluster composed of H2BVenus expressing Prdm14mut/HV rat PGCs and mEGFP/nmScarlet double-positive mouse PGCs. Scale bar: 50 Ī¼m. c Immunohistochemical staining of male gonads of Prdm14mut/HV chimeric fetus at 12 days after ET (E15-15.5). mEGFP/nmScarlet/DAZL triple-positive mouse PGCs colonized in rat gonad. In contrast, no rat PGCs are present due to defect in the Prdm14 gene. Scale bar: 50ā€‰Ī¼m. d Images of adult testis. The upper testis is a control testis from Prdm14mut/HV rat, and the lower one is from Prdm14mut/HV chimeric rats with mouse ESCs. Scale bar: 5ā€‰mm. e Immunohistochemical staining of representative seminiferous tubules. The tubules filled with nmScarlet positive cells show normal spermatogenesis originate from mouse ESC-derived germ cells. Scale bar: 50ā€‰Ī¼m. f Mouse ESC-derived sperm collected from the epididymis of Prdm14mut/HV chimeric rats. Scale bar: 50ā€‰Ī¼m. g Postnatal day 3 neonates generated by ROSI of mouse ESC-derived round spermatids into wild-type mouse oocytes. An open arrowhead shows control Slc:ICR neonate. h Images of the adult testis. The upper testis is a control testis from Prdm14mut/HV rat, and the lower one is from Prdm14mut/HV chimeric rats with mouse iPSCs. Scale bar: 5ā€‰mm. i Immunohistochemical staining of representative seminiferous tubules. The tubules filled with EGFP-positive cells show normal spermatogenesis originate from mouse iPSC-derived germ cells. Scale bar: 50ā€‰Ī¼m. j Postnatal day 1 neonates generated by ROSI of mouse iPSC-derived round spermatids into wild-type mouse oocytes. An open arrowhead shows control Slc:ICR neonate, and arrowheads show EGFP negative siblings. k A representative genotyping result to detect the CAG-EGFP transgene and retroviral integration of a reprograming factor, pMXs-Oct3/4, used for the establishment of iPSCs.

Back to article page