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PubMed1989

Optical constancy of the chick lens during pre- and post-hatching ocular development.

Embryonic and post-embryonic development of the ocular lens is associated with the continual production of new secondary lens fibers by the mitotic activity of equatorial epithelial cells. This continual development affects lens size and shape and refractive index distribution. Study of embryonic lens optical function has been largely ignored. The optical characteristics of the developing chick lens, including paraxial and eccentric focal lengths, were measured during the embryonic period of development and up to 15 days after hatching. Measurements were made with an automated scanning laser system in which the video image of a helium-neon laser beam refracted by an excised lens in solution is digitized. Focal length is measured for beams moving in small steps on either side of that center. Measurements were made on excised lenses as well as with the lens in situ within the anterior segment of the eye. The results, collected from a study of a total of 80 lenses, indicates that embryo lenses at 6-7 days of incubation have long and very variable focal lengths. At the tenth embryo day, focal length drops by more than one-half and focal variations, between lenses and for different beam positions within a single lens, is reduced. Further measures for 14- and 17-day embryo lenses, as well as for lenses from hatchling and 5-, 10- and 15-day-old chicks, indicate that there is little change in focal length, either paraxially or for eccentric beam positions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

Embryonic development of Xenopus studied in a cell culture system with tissue-specific monoclonal antibodies.

An in vitro microculture system of early gastrula cells of Xenopus laevis has been developed; deep layer cells from the lateral marginal zone (prospective somite region) or ventral ectoderm (prospective epidermis region) were fully dissociated, and the desired number of each (1-100) was distributed into a microculture well and cultured under appropriate conditions. When examined with the tissue-specific Mabs (Mu1 for muscle and E3 for epidermis, respectively), a substantial portion of the deep layer cells from the two regions followed their respective normal embryonic fates. It was found that reproducible cellular differentiation was dependent on the intimate reaggregation of dissociated cells and on the size of the resultant aggregate. About 20 lateral marginal zone cells were found to be sufficient, when put into a culture well, for supporting successful muscle differentiation, whereas about 100 ventral ectoderm cells were necessary for epidermal differentiation.

Animals↗

PubMed2006

Heparin-binding epidermal growth factor (HB-EGF) may improve embryonic development and implantation by increasing vitronectin receptor (integrin alphanubeta3) expression in peri-implantation mouse embryos.

PURPOSE: This study investigated the effects of HB-EGF on expression of integrin alphanubeta3 and implantation of embryos. METHODS: Two-cell embryos were recovered and cultured with or without 10 ng/mL HB-EGF for 96h. Expression of integrin alphanubeta3 in cultured embryos was examined by real time-RT-PCR and immunofluorescence analysis; embryos were cultured with or without HB-EGF, then transferred into the uteri of pseudo-pregnant female mice in order to analyze their implantation rate. RESULTS: HB-EGF improved embryonic hatching and outgrowth during extended culture, and up-regulated expression of integrin alphanubeta3 in both the preimplantation embryo and outgrowing blastocyst. Also, integrin alphanubeta3 subunits were localized at the pericellular borders and cell-cell contact areas. The number of successful implantation sites of transferred HB-EGF-treated embryos in the uterus was increased when compared to number of implantation sites with non-treated controls. CONCLUSIONS: HB-EGF may improve implantation by accelerating expression of integrin alphanubeta3 in peri-implantation mouse embryos.

Animals↗

PubMed2001

Embryonic development in the eggs of the silkworm, Bombyx mori, exposed to the space environment.

To investigate the effects of cosmic radiation and microgravity on embryogenesis and organogenesis in Bombyx eggs, two different stages of eggs, the early stage after oviposition and the diapause-terminated eggs, were loaded on the US Space Shuttle/Atlantis (STS-84) for a 9 day flight. More than 85% of the early stage eggs hatched in the flight sample and the ground control. In the diapause-terminated eggs, the percentage of unhatched eggs were 43% in the ground control and 56% in the flight sample. In these eggs, uncompleted embryonic reversal was observed two-fold higher percentage in the flight sample than in the ground control. The incidence of abnormality such as the larvae with segmental fusion and the appearance of abnormal crescent marking in the flight sample was significantly higher than that in the ground control. This was also observed in the 1st and 2nd filial generation of the flight sample. From these results, unsuccessful blastokinesis and the abnormal appearance was discussed in relation to cosmic radiation and microgravity.

Animals↗

PubMed2004

string(cdc25) and cyclin E are required for patterned histone expression at different stages of Drosophila embryonic development.

Metazoan replication-dependent histone mRNAs accumulate to high levels during S phase as a result of an increase in the rate of histone gene transcription, pre-mRNA processing, and mRNA stability at the G1-S transition. However, relatively little is known about the contribution of these processes to histone expression in the cell cycles of early development, which often lack a G1 phase. In post-blastoderm Drosophila embryos, zygotic expression of the stg(cdc25) phosphatase in G2 activates cyclin/cdc2 kinases and triggers mitosis. Here we show that histone transcription initiates in late G2 of cycle 14 in response to stg(cdc25) and in anticipation of S phase of the next cycle, which occurs immediately following mitosis. Mutation of stg(cdc25) arrests cells in G2 and prevents histone transcription. Expression of a mutant form of Cdc2 that bypasses the requirement for stg(cdc25) activates histone transcription during G2 in stg(cdc25) mutant embryos. Thus, in these embryonic cycles, histone transcription is controlled by the principal G2-M regulators, string(cdc25), and cdc2 kinase, rather than solely by regulators of the G1-S transition. After the introduction of G1-S control midway through embryogenesis, histone expression depends on DNA replication and the function of cyclin E, and no longer requires stg(cdc25). Thus, during the altered cell cycles of early animal development, different cell cycle mechanisms are employed to ensure that the production of histones accompanies DNA synthesis.

Animals↗

PubMed1999

The influence of maternal hypothyroidism and radioactive iodine on rat embryonal development: thyroid C-cells.

There have been no works devoted to the study of the influence of (131)I and maternal (131)I-induced hypothyroidism on the state of the C-cells in the thyroid gland of the developing embryos. A study was made on the effect of a dose of 150 microCi (131)I (0.5 Gy) leading to hypothyroidism in rats, on 35 mother rats and 168 newborn pups. The mother rats were divided into control and four treated groups which were injected with (131)I before pregnancy, on gestation days 5, 10, and 16, respectively. Immunohistochemically, the thyroid gland was examined for calcitonin-positive cells. Maternal hypothyroidism induced by (131)I leads to the development of hyperplasia and hyperthrophy of calcitonin-positive cells in the pups at the time of birth. The discovery of separate C-cells in the peripheral zone of the thyroid lobe may be evidence of an unbalance in the development of the medial and lateral source of the thyroid. There is a verifiable increase in the quantity of C-cells per 1 mm(2) field of the localization in the central zone of the gestation days 10 and 16 groups. This might be a compensatory mechanism for regulating the activity of the thyroid gland under induced hypothyroidism. Thus, in cases when there is a breakdown in the normal external regulation of the embryonic morphogenesis, a reduction in the level of maternal thyroid hormones and also direct exposure to (131)I, there is also a change in the foetus' internal regulatory systems. A change in C-cell system could lead to the appearance of endocrinological disorders later in life.

Animals↗

PubMed2004

The germ line lineage in ukigori, Gymnogobius species (Teleostei: Gobiidae) during embryonic development.

In order to determine the origin and migration of ukigori primordial germ cells (PGCs), we observed the aggregation of vasa mRNA by whole mount in situ hybridization. To observe PGC migration in the germ layers, we analyzed HE-stained paraffin sections. The germ line lineages were derived from the edge of the first, second and third cleavage furrows. During subsequent cleavages, vasa mRNA aggregations were respectively taken into four to eight cells in each embryo and vasa expressing cells proliferated from the sphere stage. At the bud to early somitogenesis period, PGCs aligned from head to tail bud regions on both sides of the embryonic body. During the late somitogenesis period, PGCs mainly aggregated just underneath the body axis. After gut formation, PGCs aligned along both sides of the gut at the 4th- to 8th- somite regions. Finally, PGCs reached the genital ridge via the inside of the lateral plate mesoderm and dorsal peritoneum. These results suggest that localized patterns of vasa transcripts and the migration routes of PGCs are different among fish (Teleost) species, perhaps depending on the amount of germinal cytoplasm derived maternally and the timing of endoderm differentiation.

Animals↗

PubMed1992

Selective dependence of mammalian dorsal root ganglion neurons on nerve growth factor during embryonic development.

We have investigated the NGF dependence of dorsal root ganglion (DRG) neurons in mammals using a paradigm of multiple in utero injections of a high titer anti-NGF antiserum. We have determined the specificity of our antiserum in relation to other members of the NGF neurotrophin family and found no cross-reactivity with brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3). To identify various classes of DRG neurons, we have stained their characteristic central projections with Dil. We show here that the NGF dependence of DRG neurons is strikingly selective. Although a majority of DRG neurons are lost after NGF deprivation during embryonic life, these are almost exclusively small diameter neurons that project to laminae I and II of the dorsal horn and presumably subserve nociception and thermoreception. Larger neurons that project to more ventral spinal laminae and subserve other sensory modalities do not require NGF for survival. These NGF-independent DRG neurons likely require one of the more recently identified neurotrophins, BDNF or NT-3.

Afferent Pathways↗

PubMed1991

Delayed fertilization and poor embryonic development associated with impaired semen quality.

Delayed fertilization is common in cycles with immature oocytes. This phenomenon was observed in 42 of 423 with mature oocytes. Of the 42 cycles, 16 were excluded because of the presence of sperm autoantibodies. Sperm parameters of the remaining 26 (6.1%) cycles (group A) were compared with those in cycles with no fertilization at all (group B) and those in the control group (group C). The percentage of normal forms was 15% in group B and 24% in group A compared with 51% in group C. Fertilization rates were 32% in group A compared with 81% in group C. The incidence of poor embryonic morphology was 82% in group A compared with 29% in group C. Delayed fertilization and poor embryonic morphology associated with impaired sperm quality is of clinical and prognostic importance.

Blastomeres↗

PubMed1987

Stimulation of early embryonic development in the sheep by co-culture with oviduct epithelial cells.

To examine the effects of somatic cell support on the cleavage and viability of fertilized sheep eggs, 434 pronucleate eggs were co-cultured for 3 or 6 days on oviduct cells or fibroblasts and 77 eggs were cultured in medium alone. During the first 3 days in culture 95% of the single-celled eggs cleaved regularly to non-compacted morulae on either of the feeder-layers but only 13% underwent similar regular cleavage in medium alone. Despite the identical cleavage rates in the co-culture groups, only 33% of embryos grown on fibroblasts as compared with 80% of embryos grown on oviduct cells were fully viable as judged by their ability to develop normally after transfer to recipient animals. The viability of embryos in the oviduct group was equal to that obtained after the direct transfer of morulae from donor to recipient sheep. After 6 days in culture 42% of embryos co-cultured with oviduct cells developed into expanded blastocysts as compared with only 4.5% cultured on fibroblasts. In both co-culture groups virtually all the remaining embryos blocked during the 4th cleavage. When transferred, 30% of blastocysts grown from the pronucleate stage on oviduct cells were viable. We conclude that: (1) during the first 3 days after fertilization cleavage will progress at a normal rate on different feeder-layers but oviduct cells appear to be required for the acquisition of full embryonic viability.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1982

Early embryonic development in the rat after maternal treatment with PGE2.

Pregnant Sprague-Dawley rats were treated subcutaneously with PGE2 at doses of 50, 100, and 200 microgram on gestational days five through eight. Control animals received the solvent in a similar manner. On gestational day 9 the animals were killed by intracardiac perfusion with Karnovsky's fixative, uteri were removed, and a transverse central slice from each implantation site was embedded in araldite. The conceptus was located and 2-3 micron sections were made of each implantation site and examined at the light microscopic level. No significant structural changes were induced in any of the conceptuses, but an unusually wide range of embryonic stages was detected in both the control and the experimental groups of animals.

Animals↗

PubMed1985

Unsaturated fatty acid compositional changes and desaturation during the embryonic development of the chicken (Gallus domesticus).

An investigation has been made to correlate the activities of the delta 9- and delta 6-long chain fatty acid desaturation systems with the increased levels of oleic and arachidonic acids in the liver relative to the yolk sac membrane of the chick embryo during the last week of development. The membrane exhibited high levels of both stearic and linoleic acid desaturation in the early stages of yolk lipid mobilization, the activities of both enzyme systems decreasing with the approach of hatching. Stearic acid desaturation in the liver also decreased with the approach of hatching, but linoleic acid desaturation increased. The observed levels of desaturation in the yolk sac membrane are capable of making a considerable contribution to the accumulations of mono- and polyunsaturated fatty acids in the embryonic liver, the requirement for which does not appear to be satisfied by the yolk lipids. With the approach of hatching and the functional regression of the yolk sac membrane, the role is taken over by the embryonic tissues.

Animals↗

PubMed1992

Teratogenicity of all-trans retinoic acid during early embryonic development in the cynomolgus monkey (Macaca fascicularis).

The embryotoxic and teratogenic potential of all-trans retinoic acid was assessed following exposure prior to and during early organogenesis in the cynomolgus monkey (Macaca fascicularis). Sixteen pregnant females were orally administered all-trans retinoic acid (Tretinoin, Hoffmann-La Roche) once daily from GD 10-20 and twice daily from GD 21-24 at three different dosages, 5 (n = 9), 10 (n = 6) and 20 mg/kg (n = 1). Adverse clinical signs resembling hypervitaminosis A were observed in one animal at 5 mg/kg, in three animals at 10 mg/kg, and in the animal treated with 20 mg/kg all-trans retinoic acid. Maternal weight loss was observed in the 10- and 20-mg/kg groups. A dose-dependent increase in embryolethality was observed, with 22% (2/9), 50% (3/6), and 100% (1/1) occurring at 5, 10, and 20 mg/kg, respectively. The majority of embryonic deaths occurred between GD 16 and 20; the incidence of these early losses was higher than in historical and concurrent controls. No malformations, but a single growth-retarded fetus, was observed in the 5-mg/kg group. Craniofacial malformations, consisting of external ear defects, mandibular hypoplasia, cleft palate, and temporal bone abnormalities, were seen in three viable fetuses in the 10-mg/kg group. Skeletal variations were common to the majority (70%, 7/10) of viable fetuses in both dose groups and were increased relative to historical controls (32%, 25/77). Unlike previous studies with 13-cis-retinoic acid during the pre- and early organogenic stages of development (Hummler et al., Teratology 42:263-272, 1990), no thymic hypo- or aplasia or heart anomalies were observed, which may be attributable to the slightly longer 13-cis retinoic acid treatment period, i.e., GD 10-27. However, external ear and temporal bone defects were common to both all-trans and 13-cis retinoic acid. The similarity observed in the malformation syndrome induced by both all-trans and 13-cis retinoic acid in the cynomolgus monkey and 13-cis retinoic acid embryopathy in humans supports this macaque species as a model for further developmental toxicity studies of vitamin A-related compounds.

Abnormalities, Drug-Induced↗

PubMed1993

Coordinate and noncoordinate regulation of synaptic vesicle protein genes during embryonic development.

Because the formation of a mature nerve terminal requires the accumulation of large quantities of synaptic vesicles, the expression of synaptic vesicle proteins would be expected to correlate with synaptogenesis. However, previous studies have provided conflicting evidence on this point. We have examined the developmental pattern of expression of mRNA and protein for three RNAs derived from two genes coding for synaptic vesicle membrane proteins. For these experiments, we cloned a chick p65 (synaptotagmin) cDNA using a reduced stringency screen with a rat p65 cDNA probe. We examined p65 expression in chick forebrain in conjunction with that of synaptophysin II. RNase protection assays for p65 and the two isoforms of synaptophysin II (Bixby, 1992) show essentially coordinate increases of these three mRNAs in embryonic forebrain during the peak period of synaptogenesis (E17 to E20). However, each of the three mRNAs has a distinct temporal pattern of expression during the early stages of embryogenesis. In the ciliary ganglion, upregulation of synaptophysin II mRNA correlates very well with synaptogenesis. Our results suggest that the regulation of expression of vesicle membrane protein mRNA can serve as a marker for synaptogenesis, despite temporal differences in early expression patterns. In contrast to mRNA expression, assays for vesicle protein expression show a relatively steady rise in both p65 and synaptophysin II throughout the embryonic period, without a sharp increase corresponding to that seen in message levels. These results suggest that the expression both of the p65 and of the synaptophysin II proteins is post-transcriptionally regulated.

Amino Acid Sequence↗

PubMed1998

The Tomaj mutant alleles of alpha Tubulin67C reveal a requirement for the encoded maternal specific tubulin isoform in the sperm aster, the cleavage spindle apparatus and neurogenesis during embryonic development in Drosophila.

The three dominant TomajD and their eleven revertant (TomajR) alleles have been localized to the alpha Tubulin67C gene of Drosophila melanogaster. Although the meiotic divisions are normally completed in eggs laid by TomajD/+, TomajD/-, TomajR/- females, embryogenesis arrests prior to the gonomeric division. The arrest is caused by: (1) the failure of prominent sperm aster formation; and (2) a consequent lack of female pronuclear migration towards the male pronucleus. Concomitant with the sperm aster defect, the four female meiotic products fuse (tetra-fusion), similar to what is seen in eggs of wild-type virgin females. In eggs of females heterozygous for weaker TomajR alleles, embryogenesis comes to a cessation before or shortly after cortical migration of cleavage nuclei. The apparent source of embryonic defect is the cleavage spindle apparatus. One of the three TomajD alleles is cold-sensitive and its cold-sensitive period coincides with the completion of female meiosis and pronuclear migration. Disorganized central and peripheral nervous systems are also characteristic of embryos derived from the temperature-sensitive TomajD/+ females. The Tomaj mutant phenotypes indicate an involvement of the normal alpha Tubulin67C gene product in: (1) the formation of the sperm aster; (2) cleavage spindle apparatus formation/function; and (3) the differentiation of the embryonic nervous system. The TomajD alleles encode a normal-sized alpha Tubulin67C isotype. Sequence analyses of the TomajD alleles revealed the replacement in different positions of a single negatively charged or neutral amino acid with a positively charged one. These residues presumably identify important functional sites.

Alleles↗

PubMed1984

Ultrasonography for detection of pregnancy and study of embryonic development in heifers.

Real-time B-mode ultrasonography was used to evaluate uterine changes associated with the estrous cycle in 22 ovulatory periods in 12 nulliparous heifers. Irregular, nonechogenic (black) areas were seen on the images of uterine horns during the periovulatory period. These nonechogenic areas were presumably due to intraluminal fluids since they coincided with the discharge of clear, viscous mucus preceding ovulation and blood-tinged mucus after ovulation. Eight heifers were bred until five pregnant heifers were obtained for study of the ultrasonic morphology of the conceptus. Ultrasound examinations were done daily to day 50 of pregnancy. Discrete, nonechogenic areas were first visible within the uterus between days 12 and 14, when they were approximately 2 mm in diameter. These discrete nonechogenic structures were identified as the embryonic vesicle, since they were observed only in heifers later confirmed to be pregnant and were always in the uterine horn ipsilateral to the corpus luteum. The presence of an embryo within the embryonic vesicle was confirmed by observing an echogenic (white) area with rhythmic pulsations (heartbeat). The embryonic vesicle gradually increased in length from the day of first observation until day 26 when it extended past the curvature of the horn and began to encroach into the contralateral horn. In all heifers, by day 32 the vesicle extended to the tip of the contralateral horn. The embryo was first visible between days 26 and 29 when the mean length was 10 mm. The embryo increased in length an average of 1.1 mm per day. A heartbeat was detectable in the embryo on the first day observed. In one superovulated heifer, five vesicles were visible in the uterine horns by day 14 and by day 33 seven embryos were observed; two of the seven embryos apparently resorbed by day 43.

Journal Article↗

PubMed1987

The Golgi apparatus in the acinar cells of the developing embryonic pancreas: II. Localization of lectin-binding sites.

The reaction patterns of the Golgi apparatus following staining with the lectins concanavalin A (ConA), Ricinus communis I agglutinin (RCA I), and Helix pomatia lectin (HPA) were studied in the pancreas acinar cells of rat embryos in the course of cell differentiation from day 13 through day 20 of gestation. The binding reactions were localized by means of pre-embedment incubation of 10-microns-thick cryosections of pancreas tissue, prefixed in a mixture of 4% formaldehyde/0.5% glutaraldehyde, using horseradish peroxidase for electron microscope visualization. ConA, which preferentially binds to alpha-D-mannosyl residues, consistently stained the cisternae of the cis Golgi side. The majority of the stacks also showed ConA staining of medial cisternae. The reaction of the trans side was variable; in each stage of development, the cisternae of the trans Golgi side either were devoid of labeling or appeared intensely stained. The reactions obtained with RCA I, which recognizes terminal beta-D-galactosyl residues, changed in the course of cell differentiation; in the protodifferentiated and early differentiated states, the system of "rigid lamellae," located at the trans side of the Golgi stacks, was intensely labeled, but became unreactive after production of secretion granules had started, the reaction then being restricted to the stacked saccules. In regard to the Golgi stacks in each of the developmental stages, RCA I binding sites either were confined to the trans cisternae, or, in addition, were found distributed across elements of the medial and cis compartments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2001

Embryonic development of the nervous system of the temnocephalid flatworm Craspedella pedum.

The nervous system of temnocephalid flatworms consists of the brain and three pairs of longitudinal connectives extending into the trunk and tail. The connectives are crosslinked by an invariant number of regularly spaced commissures. Branches of the connectives innervate the tentacles of the head and the sucker organ in the tail. A set of nerve rings encircling the pharynx and connected to the brain and connectives constitute the pharyngeal nervous system. The nervous system is formed during early embryogenesis when the embryo represents a multilayered mesenchymal mass of cells. Gastrulation and the formation of separate epithelial germ layers that characterize most other animal groups are absent. The brain arises as a bilaterally symmetric condensation of postmitotic cells in the deep layers of the anterior region of the embryonic mesenchyme. The pattern of axon outgrowth, visualized by labeling with anti-acetylated tubulin (acTub) antibody, shows marked differences from the pattern observed in other flatworm taxa in regard to the number of neurons that express the acTub epitope. Acetylated tubulin is only expressed in neurons that form long axon tracts. In other flatworm species, such as the typhloplanoid Mesostoma and the polyclad Imogine, which were investigated by us with the acTub antibody (Hartenstein and Ehlers [2000] Dev. Genes Evol. 210:399-415; Younossi-Hartenstein and Hartenstein [2000] Dev. Genes Evol. 210:383-398), only a small number of "pioneer neurons" become acTub positive during the embryonic period. By contrast, in temnocephalids, most, if not all, neurons express acTub and form long, large-diameter axons. Initially, the brain commissure, pharyngeal nerve ring, and the connectives are laid down. Commissural tracts and tentacle nerves branching off the connectives appear later. We speculate that the precocious differentiation of the nervous system may be related to the fact that temnocephalids move by muscle action, and possess a massive and complex muscular system when they hatch. In addition, they have muscular specializations such as the anterior tentacles and the posterior sucker that are used as soon as they hatch. By contrast, juveniles of Mesostoma and larvae of polyclads move predominantly by ciliary action that may not require a complex neural circuitry for coordination.

Animals↗