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PubMed2002

The SQV-1 UDP-glucuronic acid decarboxylase and the SQV-7 nucleotide-sugar transporter may act in the Golgi apparatus to affect Caenorhabditis elegans vulval morphogenesis and embryonic development.

Recent findings indicate that glycosaminoglycans can play important roles in animal development. The genes sqv-3, -7, and -8, which are necessary for vulval morphogenesis in Caenorhabditis elegans, affect the biosynthesis of chondroitin and heparan sulfate glycosaminoglycans. We cloned sqv-1 and showed that the SQV-1 protein is a type II transmembrane protein that functions as a UDP-glucuronic acid decarboxylase. SQV-1 localizes to punctate cytoplasmic compartments and colocalizes with the SQV-7 nucleotide-sugar transporter, which probably acts in the Golgi apparatus. SQV-1 and SQV-7 are both expressed in the vulva and in oocytes, where they likely act in vulval morphogenesis and embryonic development, respectively. Progeny of sqv-7 and sqv-1 null mutants fail to initiate cytokinesis, possibly because they are unable to separate the plasma membrane from the eggshell, a defect analogous to that of incomplete vulval invagination.

Amino Acid Sequence↗

PubMed2001

Cloning and characterization of a novel form of mouse fibroblast growth factor-1 (FGF-1) mRNA, FGF-1.G: differential expression of FGF-1 and FGF-1.G mRNAs during embryonic development and in postnatal tissues.

The fibroblast growth factor-1 (FGF-1) gene is characterized by the presence of different untranslated exons in its 5' end that direct the expression of alternatively spliced mRNA variants (1.A, 1.B and 1.C) that encode for FGF-1. We have previously isolated a new mouse FGF-1 upstream untranslated exon, which we termed -1G. Here we report on the cloning and characterization of the FGF-1 mRNA isoform arising from -1G. This newly identified FGF-1 mRNA species (FGF-1.G), whose transcription start site maps 295 bp upstream from the splice donor site, is predominantly expressed in young liver and kidney, where it comprises 40.2% and 30.7%, respectively, of the total FGF-1 mRNA. While the FGF-1 mRNA comprising all of the FGF-1 transcripts was present in distinct tissues at embryonic days E12.5 and E15.5, the FGF-1.G mRNA was not detected during murine embryogenesis; therefore the role of FGF-1 in embryonic development must be attributed to FGF-1 mRNAs arising from upstream untranslated exons other than -1G. On the other hand, the parallel decrease of both FGF-1 and FGF-1.G mRNA levels we observed in the aging mouse kidney and liver suggests a role of FGF-1.G in normal cellular maintenance and survival.

Aging↗

PubMed2004

Regulatory elements controlling Ci-msxb tissue-specific expression during Ciona intestinalis embryonic development.

The msh/Msx family is a subclass of homeobox-containing genes suggested to perform a conserved function in the patterning of the early embryo. We had already isolated a member of this gene family (Ci-msxb) in Ciona intestinalis, which has a very complex expression pattern during embryogenesis. To identify the regulatory elements controlling its tissue-specific expression, we have characterized the gene structure and the regulatory upstream region. By electroporation experiments, we demonstrated that a 3.8-kb region located upstream of the gene contains all the regulatory elements able to reproduce its spatial expression pattern. Analyzing progressively truncated fragments of this region, three discrete and separate regions driving LacZ reporter gene expression in the ventral epidermis, primordial pharynx and neural territories have been identified. We further investigated the element(s) necessary for Ci-msxb activation in the nervous system during embryonic development by in vivo and in vitro experiments. Both electroporation and gel-shift assays of overlapping wild type and mutated oligonucleotides demonstrated that a unique sequence of 30 bp is involved in Ci-msxb neural activation from neurula to larva stage. This sequence contains consensus binding sites for various ubiquitous transcription factors such as TCF11 whose possible implication in formation of the regulatory complexes is discussed.

Animals↗

PubMed2000

Phenoloxidase, a marker enzyme for differentiation of the neural ectoderm and the epidermal ectoderm during embryonic development of amphioxus Branchiostoma belcheri tsingtaunese.

The development of phenoloxidase during amphioxus embryogenesis was spectrophotometrically and histochemically studied for the first time in the present study. It was found that (1) PO activity initially appeared in the general ectoderm including the neural ectoderm and the epidermal ectoderm at the early neurula stage but not in the mesoderm or the endoderm, and (2) PO activity disappeared in the neural plate cells but remained unchanged in the epidermal cells when the neural plate was morphologically quite distinct from the rest of the ectoderm. It is apparent that PO could serve as a marker enzyme for differentiation of the neural ectoderm from the epidermal ectoderm during embryonic development of amphioxus.

Animals↗

PubMed1987

Progesterone receptors in hypothalamus and pituitary during the embryonic development of the chick: regulation by sex steroid hormones.

The distribution of progesterone receptor (PR) was immunohistochemically investigated in the hypothalamus and pituitary of the chick embryo (4-14 days of incubation) using an antibody against PR. PR first appears in the diencephalic floor on day 4 of incubation. From day 7 a distribution similar to that found in adult animals is observed. Between days 7 and 14, PR is detected in the periventricular and paraventricular regions, in the preoptic area and the organum vasculosum lamina terminalis. PR is not detectable in the pituitary at any stage studied. Estradiol and testosterone, but not dihydrotestosterone, increase the number of immunoreactive neurons in the hypothalamus from day 4 onwards and induce PR in the pituitary, from day 8 onwards. The early presence of PR in the brain, and its induction by estradiol in the anterior pituitary suggest a role for progesterone and estradiol in the ontogenic process of neuroendocrine regulation, very early during embryonic development.

Animals↗

PubMed2000

Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori.

Recently, it was confirmed that embryos derived from diapausing eggs of the silkworm, Bombyx mori, begin their development and reach larval maturity on mulberry leaves, when the naked eggs are cultured in vitro. In this study, we found that the method of embryo culture is useful for determining the physiological regulation of diapause. We show that the development of embryos derived from diapausing eggs was strongly inhibited by the addition of either sorbitol or trehalose to the culture medium. Furthermore, this inhibitory effect disappeared when the embryos were cultured in a control medium which did not contain either sorbitol or trehalose, indicating that the inhibitory reactions caused by both substances are reversible. The minimal effective dose of either sorbitol or trehalose was approximately 0.2 M, a value similar to the in vivo concentration of sorbitol in diapausing eggs (0.2 M). Glycerol, mannitol or glucose were moderately effective for inhibition. Sorbitol present in diapausing silkworm eggs does not appear to serve as an antifreeze, but as an strong arresting factor of embryonic development. Furthermore, these results show that a decrease in sorbitol releases the embryos from diapause at the termination of diapause.

Journal Article↗

PubMed1990

The oocyte lamin persists as a single major component of the nuclear lamina during embryonic development of the surf clam.

Nuclei and nuclear lamina-enriched fractions, isolated from 1 to 5-day-old embryos of the surf clam, Spisula solidissima, contain only one major lamin protein, which appears to be identical to the oocyte lamin (L67), as judged by 2D IEF/SDS PAGE, reactivity with a polyclonal antibody directed against L67 and 125I tryptic peptide mapping. The same protein is also present in liver, muscle, nerve and testis from adult animals. No proteins--recognized by several poly- and monoclonal antibodies, specific for somatic lamins from different vertebrate species or the oocyte lamin LIII of Xenopus- have been detected in nuclei or NL-enriched preparations, isolated from embryos or adult tissues. Synthesis of L67 is detectable in embryos 2h after fertilization; it reaches a maximum in 6h-old embryos and gradually declines thereafter. These results argue that the composition of the NL bears no obvious relationship to the structural and functional changes that take place during the embryonic development of this invertebrate.

Animals↗

PubMedDate not supplied

In vitro fertilization and embryonic development of oocytes fertilized by sperm treated with 2-deoxyadenosine.

OBJECTIVE: To test the effect in an assisted pregnancy program of an agent known to enhance sperm motility and oocyte penetration ability. DESIGN: Prospective with internal control. SETTING: Hospital clinic. PATIENTS AND INTERVENTIONS: Forty-two oocytes obtained from women undergoing IVF were inseminated with their husband's sperm washed, incubated, and capacitated in Earle's medium supplemented with 1 mM, 2.5 mM, or 5 mM 2-deoxyadenosine (2-DXA). The outcome of insemination was compared with those of 234 control oocytes from the same women that were inseminated with husband's sperm washed in 2-DXA-free medium. 15, 14, and 13 oocytes were inseminated with sperm washed in 1 mM, 2.5 mM, and 5 mM 2-DXA, respectively. RESULTS: Fertilization rates in the respective groups were 93%, 85.7%, and 85.5%. These were higher than the 70.1% fertilization rate in the control group, but only statistically higher (P < .002) in the 1 mM 2-DXA group. Embryonic development in all three 2-DXA groups was comparable to the controls. CONCLUSION: It is suggested that the value of sperm motility enhancing agents requires further evaluation in assisted pregnancy programs.

Cell Division↗

PubMed1993

Fertilization and early embryonic development in poultry.

There are two major controllers of development in the early stages of bird embryos. These are: 1) gravity, probably acting through the distribution of yolk and its components, which lays down the initial plans for polarity that are later established firmly through the genes; and 2) the primitive streak, which controls the orderly ingression of the cells and imposes a pattern on the developing tissues.

Animals↗

PubMed1999

Embryonic development of pineal gland vesicles: a morphological and morphometrical study in chick embryos.

Pineal cell aggregates in 5, 10 and 15 day-old chick embryos have been studied. Cell aggregates were classified into rosettes or vesicles and spheroid and ellipsoid vesicles distinguished. The number of pineal vesicles per unit of surface (vesicle density) was determined in three pineal portions: apical, anterior and posterior. By day 5, only cellular rosettes were found, mainly in the apical portion. After 10 and 15 days, the presence of rosettes was occasional. The posterior wall showed only small spheroid vesicles, while in the apical and anterior areas ellipsoid vesicles were also observed. Moreover, the spheroid/ellipsoid vesicle ratio increased from the 10th to the 15th day of incubation. The vesicle density decreased between the 10th and 15th day because of the increase in both vesicle and pineal size, without changes in the total number of vesicles. The results suggest that changes in vesicle morphology and density could be related to the functional activity of the pineal gland during embryonic development.

Animals↗

PubMed1995

Molecular cloning of a gene expressed during early embryonic development in Onchocerca volvulus.

Little attention has been paid to the reproductive biology of filarial nematode parasites as a possible target for immunological or chemotherapeutic intervention. An interruption of the reproductive process would, in addition to breaking the cycle of transmission, reduce the morbidity associated with certain filarial infections. As part of our efforts to define molecules that have important functions during filarial embryogenesis, antibodies against embryo-associated proteins were used to identify a 6308-bp cDNA sequence (ovt1) from an Onchocerca volvulus cDNA expression library. The ovt1 cDNA contained an open reading frame that coded for 2022 amino acids. The deduced amino acid sequence was highly hydrophilic, alpha-helical in nature and included two leucine zipper domains. OVT1 also contained a single Arg-Gly-Asp (RGD) site. The results of Southern blot analyses demonstrated that an ovt1-like gene occurs in a number of different species of filarial nematodes. In situ hybridization experiments to identify tissues that contain ovt1 transcripts showed that ovt1 was transcribed at high levels in the late morula/early blastocyst stage of embryonic development. Transcripts for ovt1 were also detected in O. volvulus larvae and in the hypodermal cells of adult parasites. Two fragments of ovt1 were expressed as fusion proteins and the fusion proteins were used to produce antibodies in rabbits. Both antibodies recognized a native protein with an apparent molecular mass of 230 kDa in extracts from gravid female O. volvulus. In addition, the antibodies reacted with a restricted number of lower-molecular mass bands which may represent the products of post-transcriptional or post-translational processing. The predicted coiled-coil structure and the sites of transcription suggest that OVT1 may be a component of the extracellular matrix.

Amino Acid Sequence↗

PubMed1999

A Caenorhabditis elegans wee1 homolog is expressed in a temporally and spatially restricted pattern during embryonic development.

A wee1 homolog, wee-1.1, is expressed in both a temporally and spatially restricted pattern during early Caenorhabditis elegans embryogenesis, and is undetectable throughout the remainder of embryogenesis. The wee-1.1 message appears to be zygotically expressed in the somatic founder cell E of the 12-cell embryo. This expression disappears when the E blastomere divides for the first time. The wee-1.1 message then appears transiently in the nuclei of the eight great-granddaughter cells of the AB somatic founder cell, just before these cells divide in the 16-cell embryo. Following this division, the wee-1.1 mRNA is no longer detectable throughout the remainder of embryogenesis. The expression of wee-1.1 in the E blastomere and in the AB progeny appears to be restricted to nuclei in prophase and metaphase of the cell cycle. Analysis of the wee-1.1 mRNA expression pattern in maternal-effect lethal mutants suggests that this expression pattern is restricted to cells of the E and AB fates in the early embryo. This mRNA expression pattern is restricted to a 10-15-min span of embryonic development and may be regulating the timing of crucial cell divisions at this early stage of development.

Amino Acid Sequence↗

PubMed2005

A Drosophila KCNQ channel essential for early embryonic development.

The mammalian voltage-dependent KCNQ channels are responsible for distinct types of native potassium currents and are associated with several human diseases. We cloned a novel Drosophila KCNQ channel (dKCNQ) based on its sequence homology to the mammalian genes. When expressed in Chinese hamster ovary cells, dKCNQ gives rise to a slowly activating and slowly deactivating current that activates in the subthreshold voltage range. Like the M-current produced by mammalian KCNQ channels, dKCNQ current is sensitive to the KCNQ-specific blocker linopirdine and is suppressed by activation of a muscarinic receptor. dKCNQ is also similar to the mammalian channels in that it binds calmodulin (CaM), and CaM binding is necessary to produce functional currents. In situ hybridization analysis demonstrates that dKCNQ mRNA is present in brain cortical neurons, the cardia (proventriculus), and the nurse cells and oocytes of the ovary. We generated mutant flies with deletions in the genomic sequence of dKCNQ. Embryos produced by homozygous deletion females exhibit disorganized nuclei and fail to hatch, suggesting strongly that a maternal contribution of dKCNQ protein and/or mRNA is essential for early embryonic development.

Animals↗

PubMed1991

Avian LHRH during embryonic development: measurement by competitive ELISA with a monoclonal antibody.

A mouse monoclonal antibody directed against chicken Gln8-luteinizing hormone-releasing hormone (cLHRH-I) was developed and characterized. This antibody was used for the development of a competitive microtiter plate enzyme-linked immunosorbent assay for avian LHRH. The assay was validated for use with tissue and was used at a working range between 5 pg and 10 ng per sample. Using this procedure, cLHRH-I and II were assayed in whole brain extracts of Japanese Quail embryos. Samples were taken at regular intervals between Day 6 of incubation through Day 1 posthatch. There were 10 samples taken at each age with 2 replicates of the entire sampling regime. Data from males and females were pooled. LHRH concentrations were low, then rose to higher levels (15 pg/mg tissue) between Days 10 through 13 and decreased thereafter. These changes are likely to be correlated with the activation of the hypothalamic-pituitary-gonadal axis. This is particularly apparent in later embryonic development.

Animals↗

PubMed1997

Inhibitory effect of human hydrosalpingeal fluid on mouse preimplantation embryonic development is significantly reduced by the addition of lactate.

Implantation and pregnancy rates following in-vitro fertilization-embryo transfer are reduced in the presence of hydrosalpinges, but the basis of the inhibition is unknown. We examined the effect of hydrosalpingeal fluid on preimplantation development of mouse embryos. Embryos cultured in 100% hydrosalpingeal fluid were significantly inhibited developmentally as compared to embryos cultured in 100% defined medium, which served as controls. In contrast, embryos cultured in 50% hydrosalpingeal fluid/50% defined medium reached the blastocyst stage at the same frequency as the controls. When lactate (final concentration 10 mM) was added to 100% hydrosalpingeal fluid, the percentage of cultured embryos that reached the blastocyst stage was significantly increased as compared to 100% hydrosalpingeal fluid, although the percentage was slightly lower than that observed for embryos cultured in the 100% defined medium. A similar but less pronounced effect occurred when pyruvate was added to hydrosalpingeal fluid. These results do not support the concept that a potent embryotoxic agent is commonly present in hydrosalpingeal fluid. Rather, they are consistent with the notion that the inhibitory effect of hydrosalpingeal fluid on embryonic development is due to the absence of essential factors, and that this deficiency can be largely corrected by the addition of energy sources.

Animals↗

PubMed2001

High-mobility group proteins 14 and 17 maintain the timing of early embryonic development in the mouse.

The high-mobility group (HMG) proteins 14 and 17 are abundant chromosomal proteins that bind to nucleosomes and enhance transcription. We report that both mRNA species and both proteins are present throughout oogenesis and preimplantation development of the mouse. When antisense oligonucleotides targeting each mRNA species are injected into one-cell embryos, the proteins become depleted at the two- and four-cell stages and reaccumulate at the eight-cell stage. One-cell embryos injected with antisense oligonucleotides targeting both HMG-14 and HMG-17 cleave to the two-cell stage. Subsequent cleavages, however, are delayed compared with control-injected embryos. Nevertheless, these embryos ultimately reach the blastocyst stage. Similarly, injection into the nuclei of two-cell embryos of a peptide corresponding to the common nucleosome-binding domain of HMG-14 and HMG-17 delays progression to the four-cell stage. Furthermore, both RNA and protein synthesis is transiently reduced in antisense-injected embryos compared with injected controls. These results identify HMG-14 and HMG-17 as constitutive components of mouse oocyte and embryonic chromatin and establish a link between the structure of embryonic chromatin and the normal progression of embryonic development.

Animals↗

PubMed2000

Vascular remnants in the rabbit vitreous body. I. Morphological characteristics and relationship to vitreous embryonic development.

Using light and transmission electron microscopy, we observed novel structures in the rabbit vitreous body. They were found in 18 out of 27 eyes from rabbits 0.5-36 months of age. These structures are scattered throughout the entire vitreous matrix. By light and transmission electron microscopy, they appear to be made up of the same structural components. Based upon their morphological appearance, they can be subdivided into two groups which we provisionally named 'intravitreal structure type 1 and 2' or 'IVS-1' and 'IVS-2'. IVS-1 has a highly variable morphology (e.g. star-shaped, round, oval), whereas IVS-2 is tubular. The dimensions of IVS-1 vary in relation to the mesh diameters of the collagen matrix, while those of IVS-2 do not. In adult rabbit eyes, we observed transitions between IVS-1 and intravitreal ghost vessels (acellular remnants of blood vessels), and between IVS-1 and IVS-2. In very young rabbits (14 days) we observed intravitreal ghost vessels consisting of tightly-packed IVS-1. Therefore, we concluded that IVS-1 and 2 are related structures presumably of vascular origin. It appears that they represent fragmented and non-fragmented acellular remnants of hyaloid blood vessels. The presence of vascular remnants throughout the entire vitreous matrix of adult rabbit eyes is in conflict with existing theories on the embryonic development of the vitreous body, which describe a strict spatial separation between the primary (vascular) and secondary (avascular) vitreous. However, it strongly supports an alternative theory that explains the formation of the secondary vitreous by a process of continuous remodelling of the primary vitreous.

Animals↗

PubMed1984

Effects of heavy ion radiation on the brain vascular system and embryonic development.

Using neonatal rats as a model system, we investigated the response of the brain vascular system to ionizing radiation and found that distinct petechial hemorrhages developed in the cerebral cortex within a few hours after irradiation, reached a maximum about 13 to 24 hours, and decreased exponentially with time. No brain hemorrhage was found in neonatal rats 12 days after irradiation. Our experimental results indicate that a dose of a few hundred rad of X rays can induce a significant number of hemorrhages in the brain, and the number of lesions increases exponentially with dose. Heavy ions induce more hemorrhages than X rays for a given dose, and the RBE for 670 MeV/u neon particles ranges from about 2.0 for low doses to about 1.4 for high doses. A histological study of the hemorrhages indicates that a large number of red blood cells leak from the blood vessels. The radiation-induced hemorrhages may be a result of some capillary membrane damages or reproductive death of some blood vessel epithelial cells. The fast onset of hemorrhage after irradiation suggest that some membrane damage may be involved. The effect of heavy-ion radiation on the embryonic development was studied with energetic iron particles. Pregnant mice were whole-body irradiated with 600 MeV/u iron particles on day 6 of gestation and were sacrificed 12 days after irradiation. Various physical abnormalities were observed, and embryos irradiated with 1 rad iron particles showed retardation of body development.

Abnormalities, Radiation-Induced↗