Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Embryos”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,585 records · Page 88Linked to original sources
PubMed2005

Preliminary investigation of the use of high frequency ultrasound imaging in the chick embryo.

BACKGROUND: The purpose of this study was to investigate the feasibility of using high frequency ultrasound to study the chick embryo in a noninvasive and longitudinal fashion. METHODS: A total of 10 SPF White Leghorn chick embryos (GDs 11-17; Hamburger and Hamilton stage 37-43) were consecutively examined with a GE Logiq 400 ProSeries ultrasound unit using an 11-MHz small parts ultrasound probe. Access for ultrasound visualization of the embryos was accomplished by opening a 2-3-cm window either in the air cell over the blunt end of the egg or laterally over the embryo-dependent side of the egg. Warmed ultrasound coupling gel was used for imaging, and thermal regulation was maintained with infant heel warmers. The ultrasound images were recorded directly on digital video using a Sony TRV 900 DV camcorder. The images were directly converted to jpeg and mjeg2 files for further analysis. RESULTS: Effective visualization of each embryo was possible on each day of the study period. The embryos were best visualized through the opening made in the air cell at the blunt end of the egg. The extent of the anatomic survey of the chick embryo was dependent upon the position of the embryo in the egg relative to the opening in the air cell. Doppler color flow mapping studies were obtained of the embryonic and extraembryonic circulation. CONCLUSIONS: This preliminary investigation clearly shows the feasibility of high frequency ultrasound imaging to study chick embryo development in a longitudinal and noninvasive fashion. Further studies are presently ongoing regarding earlier embryo development, as well as to determine the stability and dynamics of the methodology.

Animals↗
PubMed1983

Intra- and interspecific embryo transfer.

The procedures that are collectively referred to as embryo transfer (ET) have many uses. They were first used as research tools to study fetal-maternal physiology. Since the first successful mammalian embryo transfer in 1890, ET has been utilized for enhancement of genetic selection; diagnosis and treatment of infertility; control of infectious disease transmission; screening for genetic defects; propagation of rare and endangered species; and the study of developmental biology. Most of the embryo transfers have been intraspecific. A listing of the species includes rabbit, rat, sheep, mouse, goat, cattle, pig, hamster, ferret, mink, horse, baboon, cat, dog, water buffalo. In two species, rhesus monkey and humans, the successful embryo transfers have been limited to within-animal, homologous replacement of the embryos. There have been a few successful interspecific embryo transfers. The most common were between Bos taurus and B. indicus cattle. Other interspecific transfers involved Bos gaurus and B. taurus, cattle; Ovis musimon and O. aries, sheep; Equus asinus and E. caballus, horses. There are several examples of intergeneric embryo transfers in which embryos implanted but did not develop to term: sheep and goat, mouse and rat. The factors that determine the degree of compatibility between embryos and uteri of various species and genera are not clearly understood. The ability to hybridize successfully is probably a dependable indication of compatibility for embryo transfer. The limits of tolerance for differences between the donor and recipient in such factors as placental structure and gestation length have not been defined, but the recently developed technique of inner cell mass transfer will be very useful in such studies.

Animals↗
PubMed1990

Histochemical identification of primordial germ cells in diandric and digynic triploid mouse embryos.

Diandric and digynic triploid mouse embryos were isolated in the morning on day 10 of gestation. The embryos were separated from their extraembryonic membranes, and the latter were analysed cytogenetically by G-banding to establish the ploidy and sex chromosome constitution of these embryos. The diandric triploid embryos were produced by the technique of nuclear micromanipulation. Females were mated with male mice with a morphologically distinguishable "marker" chromosome to confirm the diandric status of these embryos. Digynic triploid and normal diploid embryos were isolated from LT/Sv strain females. These females spontaneously ovulate both primary and secondary oocytes, which are fertilisable and give rise to digynic triploid and normal diploid embryos, respectively. All the embryos were serially sectioned and processed in order to demonstrate the presence of alkaline phosphatase enzyme activity. This histochemical technique allowed primordial germ cells to be readily recognised, due to their characteristic location, cellular morphology, and staining appearance. Primordial germ cells were found in all the embryos studied, being located within the visceral yolk sac, at the base of the allantois, and/or in association with the wall or mesentery of the hindgut. The total number of germ cells present was established in nine diandric triploids and in five digynic triploids. The findings presented here represent the first demonstration that primordial germ cells can differentiate in either diandric or digynic triploid mammalian embryos.

Animals↗
PubMed1993

Nuclear transfer and electrofusion in bovine in vitro-matured/in vitro-fertilized embryos: effect of media and electrical fusion parameters.

In this study, micromanipulation and electrofusion conditions for the cloning of in vitro-produced bovine embryos (here after termed IVM/IVF embryos) derived from in vitro-matured (IVM) and in vitro-fertilized (IVF) oocytes were established. The effect of DC field strength on the fusion rate was tested in a model system using pronuclear stage embryos in which a cytoplasmic vesicle was removed and reinserted. Efficient fusion (80%) was obtained by applying a pulse of 1.75 kV/cm for 40 microseconds. In vitro development of manipulated pronuclear stage embryos was as efficient as that of unmanipulated control embryos. Different fusion media were compared in the cloning procedure, using IVM oocytes as recipients and blastomeres from day 6 IVM/IVF donor embryos. Zimmermann cell fusion medium reduced the lysis of nuclear transfer embryos compared to F300 (5% vs. 25%). The effects of drugs disrupting the microfilaments and microtubuli were determined. Neither the addition of cytochalasin B (CCB) for 1 hr in the postfusion medium nor incubation of donor blastomeres with nocodazole had a significant effect on the fusion or cleavage rate of the nuclear transfer embryos. Additional experiments demonstrated that there was no difference in developmental potential between nuclear transfer embryos allowed to develop in vitro or in vivo and that the embryos gave a 15% pregnancy rate in recipient cattle.

Animals↗
PubMed1989

Development in vitro of investment-free marsupial embryos during cleavage and early blastocyst formation.

Twenty embryos at the cleavage-arrested four-cell stage of the brown antechinus, Antechinus stuartii, were used to develop a method to obtain investment-free embryos by incubation at 35 degrees C in 2.5% pancreatin, 0.05% trypsin, and 0.05% pronase. Control intact embryos and the investment-free embryos were incubated in Dulbecco's Modified Eagle's Medium with high glucose and 10% foetal calf serum in 5% CO2 in air at 35 degrees C for 48 h. Release of embryos from the investments (shell, mucoid, and zona pellucida) took 4-4 1/2 h in pancreatin, 6 h in trypsin, and 55 min to 1 1/4 h in pronase. Embryonic survival in vitro was best following pronase digestion. Pronase concentrations of 0.05%, 0.1%, 0.25%, 0.5%, and 1.0% were used to free one- to 16-cell stage embryos of brown antechinus and the stripe-faced dunnart, Sminthopsis macroura, from their investments. Postincubation survival and development of embryos in vitro was best using between 0.05% and 0.25% pronase. Blastomeres in both intact and investment-free embryos shared similar characteristics during cleavage and blastocyst formation. The combined effect of these characteristics was that in intact embryos, no morula was formed, and the blastocyst developed, when cell numbers were high enough (about 32 cells), as a unilaminar structure flattened against the zona pellucida and without an inner cell mass. In investment-free embryos, the blastomeres dispersed over the base of the culture vessel. The investments were necessary to confine the blastomeres, to provide a surface for blastomere flattening, and to allow normal cellular associations to develop.

Animals↗
PubMed2007

Gene expression in bovine nuclear transfer embryos in relation to donor cell efficiency in producing live offspring.

Developmental abnormalities associated with the cloning process suggest that reprogramming of donor nuclei into an embryonic state may not be fully completed in most of the cloned animals. One of the areas of interest in this regard, is the analysis of gene expression patterns in nuclear transfer (NT) embryos to dissect the processes that failed and develop means to overcome the limitations imposed by these factors. In this study, we investigated expression patterns of histone deacetylase-1, -2, -3 (HDAC-1, -2, -3), DNA methyltransferase-3a (DNMT3A), and octamer binding protein-4 gene (OCT4) in donor cells with different cloning efficiencies and NT embryos derived from these cells employing a real-time RT-PCR assay. All genes investigated followed altered expression patterns in NT embryos when compared to IVF-derived embryos. In general, expression of HDAC genes was elevated especially at the compact morula stage and comparable to in vitro fertilized (IVF) embryos at the hatched blastocyst stage. DNMT3A expression in NT embryos was lower than IVF embryos at all stages. Oct-4 transcript levels were also reduced in cloned compared to IVF embryos at the compact morula and blastocyst stages. This difference disappeared at the hatched blastocyst stage. There was a donor cell effect on the expression patterns of all genes investigated. These results demonstrate altered gene expression patterns for certain genes, in cloned cattle embryos from our donor cells of different efficiency in producing live offspring. Therefore we suggest that differences in expression of developmentally important genes during early embryo development may characterize the efficiency of donor cells in producing live offspring.

Animals↗
PubMed1983

Teratogenic effects of valproic acid and diphenylhydantoin on mouse embryos in culture.

Teratogenic effects of the anticonvulsant drugs valproic acid (VPA) and diphenylhydantoin (DPH) on the development of mouse embryos during early organogenesis were studied using the whole embryo culture technique. Embryos with one to seven somites were exposed in vitro to 50-375 micrograms/ml VPA or 15-135 micrograms/ml DPH for up to 42 hours and compared to control embryos cultured in 80% rat serum without either drug. For both VPA- and DPH-treated embryos, a dose-dependent increase in the frequency of abnormal embryos and a decrease in viability were found. VPA and DPH produced a similar pattern of defects. Drug-induced anomalies included open neural tubes in the cranial regions, abnormal body curvature, craniofacial deformities, and yolk sac defects. Ultrastructural changes were noted in the neuroepithelium of exencephalic VPA-treated embryos. Growth and development were retarded in embryos exposed to greater than 35 micrograms/ml DPH or greater than 50 micrograms/ml VPA as indicated by the decrease in protein and DNA content and the reduction in somite number, crown-rump length, and yolk sac diameter. On a molar basis DPH was potentially more teratogenic than VPA, which correlates with the higher lipid solubility of DPH. With VPA, susceptibility to the drug depended on the developmental stage; e.g., at 150 micrograms/ml VPA the frequency of malformations was 70% in embryos with one to four somites as compared to 35% in embryos with five to seven somites.

Animals↗
PubMed1990

Effect of lithium carbonate on mouse and rat embryos in vitro.

Lithium is effective in the treatment of manic-depressive psychosis but is suspected to be a developmental toxicant in humans. It is a developmental toxicant in mice and rats in vivo, but at human therapeutic serum levels of 0.6-1.6 meq/L, rats appear to be more sensitive to the effects of the drug than do mice. The species susceptibility to lithium-induced defects was evaluated by using a rodent whole embryo culture system employing mouse and rat embryos treated at comparable developmental stages. Mouse embryos were cultured on gestational days 8-10, and rat embryos were cultured on gestational days 10-12. Care was taken to insure that all embryos had 10 +/- 2 somite pairs at the beginning of the culture period. Embryos were cultured for 44 hours in rat serum to which lithium was added to attain final drug concentrations of 0.6, 1.2, 1.8, 2.4, or 5.0 meq/L. Control embryos were treated with distilled water, which served as the vehicle. In rats, lithium induced significant decreases in various parameters at 1.8, 2.4, and 5.0 meq/L; no malformations were observed in rats of this stage. In mice, significant decreases occurred at 2.4 and 5.0 meq/L, and embryos treated at the highest concentration had a significantly increased frequency of open neural tubes. Rat embryos were also cultured with lithium on gestational days 9-11. The lowest dose producing developmental toxicity at this stage was 0.6 meq/L. Open neural tubes were present among younger rat embryos; however, this defect occurred in all groups, including the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Embryotoxic effects of chlorobutanol in cultured mouse embryos.

Chlorobutanol (CB) is a commonly used preservative which is added to numerous pharmaceutical preparations, and it is the active ingredient in certain oral sedatives and topical anesthetics. Chlorobutanol has demonstrated adverse effects in adult tissues, but CB has not been previously investigated for its effect on the developing whole embryo. The method of whole-embryo culture was used in this study to expose mouse embryos during two stages of organogenesis to CB at final concentrations of 0 (control), 10, 25, 50, 100, and 200 micrograms/ml. Embryos were evaluated for heart rate (HR), malformations, and somite number, and embryos and visceral yolk sacs (VYSs) were assayed for total protein content as a measure of overall growth. Neurulating (3-6 somite) embryos were malformed and growth retarded by exposure to CB concentrations > or = 25 micrograms/ml, with decreased VYS growth at > or = 50 micrograms/ml and decreased HR at > or = 100 micrograms/ml CB. Early limb-bud stage (20-25 somite) embryos were malformed at CB concentrations > or = 50 micrograms/ml and growth retarded at > or = 100 micrograms/ml, with decreased VYS growth at 200 micrograms/ml and decreased HR at > or = 100 micrograms/ml CB. Thus, CB produces dysmorphogenesis in mouse embryos in vitro, and neurulating embryos are somewhat less sensitive than early limb-bud stage embryos. The concentrations of CB that interfere with normal embryonic development are within the range of human blood levels measured following multiple doses of CB. Preparations containing CB should be used with caution during pregnancy, particularly when repeated dosing may allow accumulation of CB to potentially embryotoxic levels.

Abnormalities, Drug-Induced↗
PubMed1993

Strain-dependent effect of ethanol on ventricular septal defect frequency in White Leghorn chick embryos.

We have tested the potential of a single dose of ethanol (0.20 ml 50% ethanol in chick Ringer's saline (CRS) administered into the air sac) to produce ventricular septal defect (VSD) in three distinct commercially available strains of White Leghorn chick embryo: stress-resistant Dekalb Delta strain, Hy-Vac SPF type V, and Hy-Vac SPF type L. Eggs were controlled for both size and developmental stage (Hamburger-Hamilton stage 18-19) at time of injection. The frequency of VSD in Dekalb Delta embryos was 4.0% (1/25), in Hy-Vac SPF type L embryos 9.1% (3/33), and in Hy-Vac SPF type V embryos 38.9% (14/36). Statistical analysis with the two-tailed Fisher Exact Test indicated that frequencies were not significantly different (P = 0.3215) when Dekalb Delta and Hy-Vac type L embryos were compared. However, the frequency of VSD for Hy-Vac type V embryos was significantly greater than that for either the Dekalb Delta or the Hy-Vac type L embryos (P < 0.005). All VSDs observed were located within the crista supraventricularis. When Hy-Vac SPF type V embryos were exposed to either 0.20 ml 50% ethanol in CRS or to 0.20 ml CRS (controls), ethanol-treated embryos showed a VSD incidence of 34.1% compared with a 3.6% incidence in the controls (P = 0.0017). These data suggest that ethanol is the cause of VSD in this strain. From the results of this study, we are led to conclude that different commercial strains of White Leghorn chick embryo show different susceptibilities to the induction of VSD by ethanol.

Animals↗
PubMed1995

Genetically determined absence of an initiation site of cranial neural tube closure is causally related to exencephaly in SELH/Bc mouse embryos.

The SELH/Bc mouse strain (SELH) has a high frequency of the lethal neural tube closure defect, exencephaly, in newborns and embryos. Previous work has shown that all SELH mouse embryos have an abnormal mechanism of rostral neural tube closure. They lack initiation of contact and fusion of the cranial neural tube at the prosencephalon/mesencephalon boundary [Closure 2), and undergo closure by extension of a more rostral site of fusion. This process fails in 10-20% of embryos, where the mesencephalic folds remain unelevated, resulting in exencephaly. Previous work has also shown that the cause of liability to exencephaly in SELH mice is multigenic, involving a small number of loci. The purpose of the present study was to test the hypothesis that the genes causing the lack of Closure 2 also cause the liability to exencephaly in SELH, by observation of their joint transmission from genetically segregating animals. A concurrent mapping study provided the necessary genetic material, a segregating F2 generation from a cross of SELH with the normal LM/Bc strain. The genetic liability to exencephaly transmitted by individual F2 sires had been measured by the frequencies of exencephalic day 14 embryos they produced in test-crosses with SELH females. A selected subset of 13 of these test-crossed F2 sires was bred with a second set of SELH females, and the embryos were examined earlier, during the period of neural tube closure, on days 8 and 9 of gestation, to determine the presence of Closure 2. Six F2 sires were among the highest exencephaly producers (6-11%), six were among the lowest (0%), and one was intermediate (5%). Among embryos at the appropriate stage for scoring, the presence of Closure 2 was observed to be inversely correlated with the later risk of exencephaly, being present in 93% (71/76) from the low-risk sires and 35% (36/103) from the high-risk sires. In each case, the remaining embryos had a closure mechanism like that of SELH embryos. Among the individual intermediate- and high-risk sires, there was also a clear correlation between the frequency of exencephaly in older embryos and the frequency of lack of Closure 2 in early embryos (rs = 0.88; P < 0.05). This study demonstrates that high liability to exencephaly and absence of Closure 2 are genetically transmitted together. That is, the cause of the lack of Closure 2 in SELH mice is shown to be also the probable cause of the high liability to exencephaly.

Animals↗
PubMed1993

The use of sodium hyaluronate in freezing media for bovine and murine embryos.

Four experiments were designed to determine whether sodium hyaluronate (SH) may be used to replace newborn calf serum (NCS) in murine and bovine embryo freezing media. A total of 780 mouse and 178 cattle embryos were frozen. Freezing media were prepared in Dulbecco's phosphate-buffered saline supplemented with 20% NCS and 10% glycerol (v/v) (medium A). In experiments I and II, NCS was replaced by 0.1% SH (w/v) in medium B and by 0.1% polyvinyl alcohol (PVA) (w/v) in medium C. In experiments III and IV, NCS was replaced in mouse embryo freezing medium by three different molecular weights of SH as follows: 0.1 or 0.2% (SH-1; < 3 x 10(5) Da) in medium B, by 0.1 or 0.05% (SH-2; 5-7.5 x 10(5) Da) in medium C, and by 0.1 or 0.025% (SH-3; > 1.2 x 10(6) Da) in medium D. Embryos were frozen and thawed using standard procedures. Glycerol was diluted from embryos after thawing in a single step with 1.0 M sucrose. After 48 h in culture (Ham's F-10 medium for cattle embryos and BWW medium for mouse embryos), embryos were evaluated for development to expanded or hatched blastocysts. Survival rates were compared by chi 2 analysis. In experiment I, there were no significant differences in mouse embryo survival or development between NCS, SH, and PVA groups (84.1, 79.2, and 83.3%, respectively). In experiment II, there were no significant differences in bovine embryo survival between NCS and SH groups (67.2 and 67.7%), but both had significantly higher survival than the PVA group (37.9%; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1997

New approaches for studying the permeability of fish embryos: toward successful cryopreservation.

This paper describes some new approaches for understanding the permeability of teleost embryos. The dechorionated zebrafish (Brachydanio rerio) was used as a model for basic studies of water and cryoprotectant permeability. These embryos are composed of two compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells. Cellular water was distributed unequally in each compartment. Measurements indicated that the total water in the embryo was 74%, while the total water in the yolk was 42%, and total water in the blastoderm was 82%. The internal isosmotic value for the zebrafish embryo is unknown. However, for one-compartment modeling studies of membrane permeability, the mean Lp (+/- SEM) values were 0.022 +/- 0.002 to 0.049 +/- 0.008 microns x min-1 atm-1 at 40 mOsm (assuming this was one possible internal isosmotic value for the entire embryo) and 0.040 +/- 0.004 to 0.1 +/- 0.017 microns x min-1 atm-1 at 300 mOsm (assuming this was another possible internal isosmotic value for the entire embryo). When three- and six-somite embryos were placed in 1.5 and 2.0 M cryoprotectants (dimethyl sulfoxide and propylene glycol), osmometric measurements of volume changes indicated no cryoprotectant permeation. However, similar measurements with methanol revealed a small volume decrease (ca. 8%) and recovery (ca. 5%) for six-somite embryos in a 2.0 M solution. Magnetic resonance (MR) images of the spatial distribution of three cryoprotectants (dimethyl sulfoxide, propylene glycol, and methanol) demonstrated that only methanol permeated the entire embryo within 15 min. The other cryoprotectants exhibited little or no permeation into the yolk over 2.5 h. The results from MR spectroscopy and cryoprotectant microinjections into the yolk suggested that the yolk syncytial layer plays the critical limiting role for cryoprotectant permeation throughout the embryo.

Animals↗
PubMed1998

Permeability of dechorionated one-cell and six-somite stage zebrafish (Brachydanio rerio) embryos to water and methanol.

The permeability of dechorionated one-cell and six-somite stage zebrafish (Brachydanio rerio) embryos to water and the cryoprotectant methanol at 22 degreesC was studied, using real-time video microscopy to determine the volumetric changes of the embryos during cryoprotectant exposure. The equilibrium volumetric behavior of the embryos and the Boyle-van't Hoff relationships were determined using sucrose as a nonpermeating compound. These showed the embryos to behave as nearly ideal osmometers over the range of 253-1724 mOsm, with osmotically inactive volumes of 72.9 and 82.6% for one-cell and six-somite stage embryos, respectively. The Boyle-van't Hoff relationship of the ovary eggs was also determined for comparison and their osmotically inactive volume was 63.9%. The Kedem-Katchalsky parameters of water permeability (Lp), cryoprotectant permeability (Ps), and reflection coefficient (sigma) were determined using DIFFCHAM software. The parameters reported in this study are phenomenological parameters referring to the overall embryo response. The mean values of these parameters were Lp = 0.34 and 0.35 (micrometer/min*atm), Ps = 0.45 and 0.04 (micrometer/s), and sigma = 0.88 and 0.93 for one-cell and six-somite stage embryos, respectively. While the water permeability of the dechorionated zebrafish embryos at different developmental stages remained relatively stable, the permeability to the cryoprotectant methanol (Ps) appeared to decrease during embryo development. The Ps and sigma values for methanol are the first reported for dechorionated fish embryos at these stages.

Animals↗
PubMed1996

Hensen's node from vitamin A-deficient quail embryo induces chick limb bud duplication and retains its normal asymmetric expression of Sonic hedgehog (Shh).

Both Hensen's node, the organizer center in chick embryo, and exogenous retinoic acid are known to induce limb duplication when grafted or applied to the host chick limb bud. Retinoic acid is known to be present in the node and has been proposed as the putative morphogen for chick limb development. Here, we report that Hensen's node from vitamin A-deficient quail embryo induces limb duplication in the host chick embryo similar to that induced by the node from vitamin A-sufficient control embryos. We also demonstrate that the expression of Sonic hedgehog (Shh), recently shown to be the mediator of polarizing activity in the chick limb bud, is not affected by the endogenous vitamin A status of the embryo. Furthermore, whole-mount in situ hybridization revealed asymmetry of Shh expression in the Hensen's node of both vitamin A-sufficient and -deficient quail embryos. Retinoids were not detectable in the eggs from which vitamin A-deficient embryos were obtained. Extracts from normal embryos induced a level of expression of reporter gene equivalent to the presence of 3.4 pg of active retinoids per embryo, while those from vitamin A-deficient embryos induced a baseline level of reporter gene expression similar to that of the controls. Our studies suggest that endogenous retinoic acid is not involved in Shh expression nor in regulating its asymmetry during normal early avian embryogenesis and support the current view that endogenous retinoic acid may not be a direct morphogen for limb bud duplication.

Animals↗
PubMed1999

Proliferative and steroidogenic effects of follicle-stimulating hormone during chick embryo gonadal development.

The aim of this study was to evaluate the in vitro effect of human follicle-stimulating hormone (hFSH) on cellular proliferation and steroid hormone secretion in the left ovary, the right ovary, and the testis of the chick embryo. Gonads from 8- to 18-day-old chick embryo were cultured in a defined medium during 60 h under basal and hFSH-stimulated conditions (0.5 IU/ml of culture medium). At the end of the culture, the incorporation of ¿(3)Hthymidine and the total number of cells were measured to estimate gonadal cell proliferation. The secretion of 17beta-estradiol and testosterone in the culture medium was radioimmunologically assayed in order to evaluate the steroidogenic function of the cultured gonadal cells. The response to hFSH stimulation was observed in the left ovary, the right ovary, and the testis from the 8-day-old chick embryo. In the left ovary, cellular proliferation was not augmented by hFSH in the 8-, 10-, and 13-day-old chick embryo; meanwhile, the proliferative stimulus of hFSH was observed in the 15- and 18-day-old embryos. In the same ovary, 17beta-estradiol and testosterone secretion were stimulated after hFSH treatment at all evaluated stages (8-18 days of chick embryo development). In the right ovary, an increment in proliferation and steroidogenesis was induced by hFSH in the 8-, 10-, and 13-day-old chick embryo. Afterward, the right gonad did not respond to hFSH. Testis cells displayed hFSH response as an increment in cell proliferation at all embryonic ages (8-18 days of chick embryo development). There was a transient lack of response to hFSH in testosterone secretion at 10 and 13 days of development. The in vitro effect of hFSH on cell proliferation and steroid hormone secretion changed in the ovary and the testis according to the age of the embryo. These changes could be attributed to the growth of the left ovary and the testis and the regression of the right ovary. Probably, paracrine factors modulated the gonadotropin effect on the target cells during embryonic development of chick embryo gonads.

Animals↗
PubMed1997

Analysis of heart development in cultured rat embryos.

The long-range goal of this research is to establish an in vitro system that will permit pertubation of mammalian heart development and in situ examination of the cellular and molecular events underlying cardiac morphogenesis. Rat embryos at 9.5-11.5 days of gestation were placed in culture bottles containing rat serum and Tyrode's solution. Embryos cultured for 24 and 48 h were compared to age-matched in vivo controls for morphological score, morphometric analysis of heart development, and confocal and electron microscopic analysis of myofiber pattern formation. Morphological scores indicated that embryos cultured for 24 h from day 9.5 to 10.5 had essentially normal development when compared to age-matched embryos allowed to develop in vivo. Development of embryos maintained for 48 h in culture was slightly delayed at 66-68% of age matched in vivo embryos. Analysis of hearts from embryos allowed to develop 9.5-11.5 days in vivo plus 24 and 48 h in culture showed that the ventricular thickness and height, as well as the truncal, atrial and ventricular diameters were equivalent to those of hearts from age-matched in vivo controls. Hearts from embryos allowed to develop from 11.5-12.5 days in vitro and cultured for 24 and 48 h had smaller left ventricular and atrial dimensions than controls. Cardiac myofibrillogenesis and myofibrillar pattern formation in embryos cultured from 9.5 days of in vivo development for 48 h were also normal. These studies indicate that the rat whole embryo culture system is a useful model to study several critical periods in mammalian heart development.

Animals↗
PubMed1986

Gene transfer into mouse embryos.

Gene transfer into the murine genome was accomplished nearly a decade ago by use of chimeras and teratocarcinomas; however, the low frequencies of transfer into the germ line and other difficulties stemming from mosaicism and karyotypic abnormalities in chimeric mice have limited the general usefulness of this procedure in achieving transformation in mammalian embryos. The introduction of cloned genes into teratocarcinoma cells, selection for a mutant phenotype, and transfer of those cells into mouse embryos holds some promise as a technique to employ mouse chimeras for gene transfer into mice. Infection with animal viruses and retroviral vectors provides another way to introduce exogenous DNA into mouse embryos. Infection with Mo-MuLV has been utilized to characterize the relationship between sites of integration and gene function in developing and adult mice. Gene transfer by microinjection of cloned recombinant DNA has been used by many laboratories for the transfer of DNAs into mouse embryos. The factors affecting transformation frequencies and sites of integration are unknown at present, although it seems that integration is not strictly mediated by homology-dependent events. Many genes have been introduced into mouse embryos by these procedures and many of these are expressed at high levels in appropriate tissues. No realistic possibility exists at the present time for the utilization of embryo gene transfer in the medical field for the correction of genetic defects for several reasons. First, in order to effectively provide "gene therapy" it would be necessary to determine the genotype of each recipient egg, a technical impossibility. The genetic diseases that would be amenable to germ line intervention are recessive diseases and there would be only a 25% chance of any one embryo derived from heterozygous parents being a homozygous recessive. Moreover, it would be impossible to distinguish the normal from abnormal embryos. Second, the frequencies of transformation are so low as to exclude work on human beings on ethical grounds. Third, the parameters effecting chromosomal integration sites and gene expression have not been fully characterized. Until it becomes experimentally possible to target the newly introduced DNA into expressable chromosomal sites and actively replace or supplement defective genes, the possibility of gene therapy through manipulation of embryos is remote. Yet, efforts to provide gene therapy in somatic tissues have been promising, leading to expression of a modified phenotype (Anderson, 1984). In contrast to embryo gene therapy, gene therapy in somatic tissues would not lead to germ line propagation of the manipulated genotype.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗