Late Period Of Organogenesis Research Articles

Dependence of malformation upon gestational age and exposed dose of gamma radiation.

In order to evaluate the importance of gestational age and the dose-incidence relationship by gamma radiation, pregnant ICR mice at gestational days from 2.5 to 15.5 days post-coitus (p.c.) were exposed to a single dose of 2.0 Gy and also at day 11.5 after conception, which was the most sensitive stage for the induction of major congenital malformations. The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, changes in head size and other morphological abnormalities. The only demonstrable effect of irradiation during the pre-implantation period was an increase in prenatal mortality. Resorptions were maximal on exposure at day 2.5 after conception. The pre-implantation irradiated embryos which survived did not show any major fetal abnormalities. A small head, growth retardation, a cleft palate, dilatation of the cerebral ventricle, a renal pelvis, and abnormalities of the extremities and tail after exposure were prominent during the organogenesis period, especially on day 11.5 of gestation. As for the dose-incidence relationship, the incidence of a small head, growth-retarded fetuses, a cleft palate, dilatation of cerebral ventricle and abnormalities of the extremities in live fetuses rose as the radiation dose increased. The result indicated that the late period of organogenesis in the development of the brain, skull and extremities of a mouse was a particularly sensitive phase. The threshold doses of radiation that induced a cleft palate and dilatation of the cerebral ventricle, and abnormal extremities were between 1.0 and 2.0 Gy, and between 0.5 and 1.0 Gy, respectively.

Involvement of p53-dependent apoptosis in radiation teratogenesis and in the radioadaptive response in the late organogenesis of mice.

The irradiation of fetuses at the late period of organogenesis has been known to induce a dramatic increase in malformations. The mechanisms involved, however, have remained unclear for a long time. Using the mouse limb bud system, we first found that radiation-induced apoptosis is involved in the malformation, namely, radiation-induced apoptosis in the predigital regions of embryonic limb buds is responsible for digital defects in mice. An examination of embryonic C57BL/6J mice with different p53 (trp53) status enabled us to further find that susceptibility to radiation-induced apoptosis in the predigital regions and digital defects depend on both the p53 status and the radiation dose; p53 wild-type mice appeared to be the most sensitive, while p53 knockout mice were the most resistant. These results indicate that p53-dependent apoptosis mediates radiation-induced digital defects in the later organogenesis period. The existence of a radioadaptive response in embryonic mice, which has not been reported so far, was found by irradiating embryos with either 5 cGy or 30 cGy on embryonic day 11 prior to a challenging irradiation at 3 Gy on embryonic day 12. p53-heterozygous embryos did not show the radioadaptive response, indicating the involvement of p53 in the radioadaptive response in embryogenesis.

Influence of gestational age at exposure on the prenatal effects of gamma-radiation

The objective of this investigation was to evaluate the influence of gestational age at exposure on the prenatal effects of gamma-radiation. Pregnant ICR mice were exposed to a single dose of 2.0 Gy gamma-radiation at a gestational 2.5 to 15.5 days post-coitus (p.c.). The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, change in head size and any other morphological abnormalities. The only demonstrable effect of irradiation during the preimplantation period was an increase in prenatal mortality. Resorptions were maximal on post-exposure day 2.5 after conception. The pre-implantation irradiated embryos which survived did not show any major fetal abnormalities. Small head, growth retardation, cleft palate, dilatation of the cerebral ventricle, dilatation of the renal pelvis and abnormalities of the extremities and tail were prominent after exposure during the organogenesis period, especially on day 11.5 of gestation. Our results indicate that the late period of organogenesis in the mouse is a particularly sensitive phase in terms of the development of the brain, skull and extremities.

Adaptive Response in Embryogenesis: I. Dose and Timing of Radiation for Reduction of Prenatal Death and Congenital Malformation during the Late Period of Organogenesis

An adaptive response was demonstrated during embryogenesis in mice. Whole-body irradiation at a dose of 0-50 cGy was given to condition pregnant ICR mice on day 9 to day 11 of gestation. Then their whole bodies were exposed to a challenging dose of 5 Gy on the next day. The numbers of living fetuses, prenatal deaths and living fetuses with external gross malformations were determined on day 19. A conditioning dose of 30 cGy on day 11 significantly increased the rate of living fetuses and reduced the incidence of congenital malformations induced by a 5-Gy dose on day 12. This indicates the existence of a critical dose and timing for administering a conditioning dose for radioadaptation during the late period of organogenesis in mice. The possible mechanisms involved are discussed.

Influence of gestational age at exposure on the prenatal effects of gamma-radiation.

The abdominal region of pregnant Swiss albino mice was exposed to single dose of 0.5 Gy gamma-radiation at gestation days from 1.5 to 17.5 days post-coitus (p.c). The animals were sacrificed on day 18 p.c. and foetuses were examined for resorption and embryonic death, foetal death, growth retardation, small head, low brain weight, micro-phthalmia and any other gross morphological abnormalities. The period of maximum sensitivity for each effect varied. The only demonstrable effect of irradiation during the pre-implantation period was an increase in prenatal mortality. Resorptions were maximal after exposure between days 2 and 4 p.c. The pre-implantation irradiated embryos which survived did not show any major foetal abnormalities. These results confirm earlier mouse studies using higher doses of X-rays. Small head, low brain weight and microphthalmia were prominent after exposure during the late organogenesis period, especially between days 9 and 13 p.c. But no other externally visible anomalies were detected. These findings demonstrate that mouse organogenesis is very sensitive to radiation-induced retardation of development, even at doses < 1 Gy. One exencephaly, one cleft palate and two cases of open eyelids were observed in the foetuses exposed on days 14.5 and 15.5 p.c.; the number of these cases was too small to indicate a causal relationship with exposure.

Effect of Exposure to Low-Dose Gamma Radiation during Late Organogenesis in the Mouse Fetus

The abdominal region of pregnant Swiss mice was exposed to 0.05 to 0.50 Gy of gamma radiation on day 11.5 postcoitus. The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, changes in head size and brain weight, and incidence of microphthalmia. No marked increase in fetal mortality or growth retardation was observed below 0.25 Gy; the increase in these parameters was significant only at 0.50 Gy. A significant reduction in head size and brain weight and a significant increase in the incidence of microphthalmia were observed at doses above 0.15 Gy. Detectable levels of microcephaly and microphthalmia were evident even at 0.10 Gy. A linear dose response was seen for these effects in the dose range of 0.05 to 0.15 Gy. It is concluded that the late period of organogenesis in the mouse, especially between days 10 and 12 postcoitus, is a particularly sensitive phase in the development of the skull, brain and eye.

Effect of prenatal ultrasound exposure on adult behavior in mice

Pregnant Swiss mice were exposed to diagnostic levels of ultrasound (3.5 MHz, Maximum acoustic output: I SPTP = 1 W/cm 2 and I SATA = 240 mW/cm 2, acoustic power = 65 mW) min on days 11.5 or 14.5 postcoitus (PC). At 3 and 6 months postpartum, offspring were subjected to the following behavioral tests: bright and dark arena test for locomotor/exploratory activity and passive avoidance test for learning and memory. Anxiolytic activity and latency in learning were noticed in the ultrasound-treated animals. The effect was more pronounced in the 14.5 days PC group than in the 11.5 days PC group. But memory was not affected in the ultrasound-exposed animals. There was a nonsignificant decrease in the total locomotor activity at 6 months of age in all the exposed animals. Thus, the present data demonstrate that exposure to diagnostic ultrasound during late organogenesis period or early fetal period in mice may cause changes in postnatal behavior as evidenced by selected adult offspring behavioral tests. However, any conclusive statement in this regard should await results from more detailed investigations.

Effect of in utero exposure to diagnostic ultrasound on the postnatal survival and growth of mouse.

Pregnant Swiss albino mice were exposed to 3.5 MHz diagnostic ultrasound for 10 min (upper limit for ISPTP = 1 W/cm2 and for ISATA = 240 mW/cm2, acoustic power = approximately 65 mW) on day 3.5 (preimplantation period), 6.5 (early organogenesis period), 11.5 (late organogenesis period), or 14.5 (early fetal period) of gestation. The offspring were observed for changes in litter size at birth, and sex ratio at 4 weeks of age, and postnatal mortality and growth retardation up to 6 weeks of age. No significant difference from control in litter size or sex ratio was observed in the offspring exposed to ultrasound on any of the gestation days studied. Exposure at the early organogenesis period produced a marginally significant increase in the postnatal mortality. A significant number of ultrasound exposed animals showed lesser body weights than the control group. However this growth retardation was transient and the normal growth pattern was restored by 6 weeks of age. It is concluded that the early stages in mouse gestation may be sensitive to the lethal effects of ultrasound than the later stages of pregnancy. Even though a small increase in the postnatal mortality was observed in the ultrasound exposed animals, the normal growth was not affected in the surviving animals except for a transient growth retardation. Low birth weight may be a characteristic effect of exposure only at the preimplantation period.

Effect of Prenatal Exposure to Diagnostic Ultrasound on the Development of Mice

Pregnant Swiss albino mice were exposed to diagnostic ultrasound (3.5 MHz, approximately 65 mW) for 10 min on Day 3.5 (preimplantation period), 6.5 (early organogenesis period), or 11.5 (late organogenesis period) of gestation. Sham-exposed controls were maintained for comparison. Exposed as well as control fetuses were dissected out on the 18th day of gestation, and changes in total mortality, body weight, body length, head length, brain weight, sex ratio, and microphthalmia were recorded. Exposure on Day 3.5 of gestation resulted in a small increase in the resorption rate and a significant reduction in fetal body weight. A low fetal weight and an increase in the number of growth-retarded fetuses were produced by exposure on Day 6.5 postcoitus. A statistically nonsignificant increase in the incidence of microphthalmia was induced in fetuses exposed on Day 6.5 or Day 11.5 of gestation. These results indicate that ultrasound may have some adverse effects on the mouse embryos depending on the developmental stage at which the exposure occurred.

Effect of low dose of 70 kVp X-rays on the intrauterine development of mice

Pregnant Swiss albino mice were exposed to low doses of X-rays (approximately 9 mGy) in the range used for diagnostic exposure, on day 3.5 of gestation (preimplantation period), day 6.5 (early organogenesis period) or day 11.5 (late organogenesis period). The fetuses were examined on the 18th day of gestation. Exposure at 3.5 days post coitus (d.p.c.) resulted in a significant increase in prenatal mortality, and an increased incidence of retarded fetuses was observed after exposure at 3.5 and 6.5 d.p.c. The major effect of exposure at 11.5 d.p.c. was a significant decrease in the fetal head size and brain weight.

Cytotoxic effects of ethylnitrosourea on central nervous system of rat embryos. Special references to carcinogenesis and teratogenesis.

For the purpose of observing the cytotoxic effect of N-ethyl, N-nitrosourea (ENU), a potent carcinogen on the central nervous system (CNS) at the late period of organogenesis, the embryonal telencephalic wall was hourly examined under the light and electron microscopes. Pregnant rats on the 13th day and 15th day of gestation were given either a single intravenous injection of 40 or 80 mg/kg, of ENU. The cytotoxic effect on the embryo treated on the 15th day of gestation was severer than that treated on the 13th day of gestation. The common pathological changes in both treated groups are; (1) mitotic arrest 1 hour after administration in the most inner zone of the ventricular layer, (2) degeneration and necrosis accurred predominantly in the so-called DNA synthetic zone and resulting in cell loss of the ventricular layer, (3) elimination of degenerating products; and (4) tissue repair. The findings mentioned may indicate a cytotoxic effect inducing microencephalia, and furthermore, the teratogenic and carcinogenic mechanisms are discussed.