Figure 3: The phICSI protocol applied to round spermatids: phROSI.
From: Mice produced by mitotic reprogramming of sperm injected into haploid parthenogenotes
(a) Representation of phROSI showing (top left) Hoffman modulation image of round spermatid pick-up (bar, 5 μm). (b) Histograms showing in vitro development at E5.0 following phICSI (left) and phROSI of embryos injected at the times shown (0–2 h) after pronuclear membrane breakdown (pnMBD). Data are for all embryos (open) or those expressing paternal membrane tdTomato (mtdT). Values of n are from experiments on 2 days, with differences shown for P<0.05 (χ2 test). (c) Vertically paired Hoffman (upper) and fluorescence (mtdT) images of blastocysts (E5.0) developing from parthenogenetic haploid embryos (1nP) and phROSI-pnMBD-2h. Scale bar, 100 μm. (d) Average term (P0) weights of pups (left) and placentae. For ROSI, n=16; phROSI, n=6. Experiments were on 3 days. Differences are shown where P<0.05 (two-tailed, unpaired t-test). (e) Representative images of term (P0) pups and placentae (left panels) and F1 phROSI-pnMBD-2h and control ROSI offspring following natural mating. Values in b and d are ±s.e.m. and show P<0.05.
