A novel technique was developed to deliver a
bolus dose of a DNA label into the peritoneal cavity of fetal sheep at
85–130 days gestation. Use of markers to identify the site of injection
in fetuses from litters up to quadruplets, and immunohistochemistry to detect
the DNA label, 5-bromo-2¢-deoxyuridine (BrdU), confirmed the procedure
was successful in 85% of cases. Duration of the procedure was (mean SD)
44 16 min, and recovery from anaesthesia was rapid and uneventful in all
cases. Fetal weight was estimated with a high degree of accuracy (residual
standard deviation (RSD) = 297 g and r
2 = 0.93, P<0.001) and
the dose of label administered (110 33 mg BrdU/kg fetal weight) was
adequate in all cases. BrdU detected in fetal nuclei following injection into
amniotic fluid highlights the need for positive identification of the
injection site in timed, short-term studies, and suggests potential to further
develop the technique to investigate cellular events in fetal sheep younger
than 85 days of gestation. The results demonstrate that the procedure can be
used to determine in vivo whether or not nuclei have
entered the S-phase of the cell cycle.