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PubMed2005

Gene expression in pharyngeal arch 1 during human embryonic development.

Craniofacial abnormalities are one of the most common birth defects in humans, but little is known about the human genes that control these important developmental processes. To identify relevant genes, we analyzed transcription profiles of human pharyngeal arch 1 (PA1), a conserved embryonic structure that develops into the palate and jaw. Using microdissected, normal human craniofacial structures, we constructed 12 SAGE (serial analysis of gene expression) libraries and sequenced 606 532 tags. We also performed Affymetrix microarray analysis on 25 craniofacial targets. Our data revealed not only genes "enriched" or differentially expressed in PA1 during fourth and fifth week of human development, but also 6927 genes newly identified to be expressed in human PA1. Many of these genes are involved in biosynthetic processes and have binding function and catalytic activity. We compared expression profiles of human genes with those of mouse homologs to look for genes more specific to human craniofacial development and found 766 genes expressed in human PA1, but not in mouse PA1. We also identified 1408 genes that were expressed in mouse as well as human PA1 and could be useful in creating mouse models for human conditions. We confirmed conservation of some human PA1 expression patterns in mouse embryonic samples with whole mount in situ hybridization and real-time RT-PCR. This comprehensive approach to expression profiling gives insights into the early development of the craniofacial region and provides markers for developmental structures and candidate genes, including SET and CCT3, for diseases such as orofacial clefting and micrognathia.

Animals↗
PubMed2000

Comparisons of different stages of chick embryonic development by the physiological regulatory response to hyposmotic challenge.

Cardiac myocytes isolated and cultured from 11 day chick embryos present a Ca(2+)-dependent regulatory volume decrease (RVD) when exposed to hyposmotic stimulus. The RVD of myocytes from different embryonic stages were analyzed to evaluate their physiological performance through development. Among the several embryonic stages analyzed (6, 11, 16 and 19 days) only 19 day cardiac myocytes present a greater RVD when compared with 11 day (considered as control), the other ages showed no difference in the regulatory response. As it is known that RVD is Ca(2+) dependent, we decided to investigate the transient free Ca(2+) response during the hyposmotic swelling of the 11 and 19 day stages. The 11 day cardiac myocyte showed a transient 40% increase in intracellular free Ca(2+) when submitted to hyposmotic solutions, and the free Ca(2+) returned to baseline levels while the cells remained in hyposmotic buffer. However, the intracellular free Ca(2+) transient in the 19 day cells during hyposmotic challenge increases 100% and instead of returning to baseline levels, declines to 55% above control, well after the 11 day transient has returned to baseline. Also, quantitative fluorescence microscopy revealed that 19 day cardiac myocytes have more sarcoplasmic reticulum (SR) Ca(2+) ATPase sites per cell as compared to the 11 day cells. Our findings suggest that 19 day cells have more developed intracellular Ca(2+) stores (SR). By evoking the mechanism of Ca(2+) induced Ca(2+) release, the cells have more free Ca(2+) available for signaling the RVD during hyposmotic swelling.

Animals↗
PubMed1987

Receptors for insulin-like growth factors I and II in developing embryonic mouse limb bud.

Insulin-like growth factors (IGF) or somatomedins (SM) have been classically defined as promoting the actions of growth hormone in skeletal growth. IGF is divided into two groups, IGF-I and II, and are presumed to act via IGF type I (higher affinity for IGF-I and II and very low affinity for insulin) and II (higher affinity for IGF-II than I and no affinity for insulin) receptors, respectively. Recently, a switchover role of IGF-II to I during fetal to adult growth has been suggested. We have investigated the possible transitional role of IGF-II to I in a developing mouse embryonic limb bud organ culture model. In this in vitro system, limb bud develops from the blastoma stage to a well-differentiated cartilage tissue. Both IGF type I and II receptors were found to be present in limb buds at all stages of differentiation. Type I receptor decreased with differentiation while Type II receptor increased. The effect of IGF-I on [3H]thymidine and [35S]sulfate uptake by the tissue increased with differentiation while the effect of IGF-II on [3H]thymidine uptake of the undifferentiated tissue was abolished with differentiation of the tissue. The increase of the IGF-I response with decreased type I receptor may reflect an altered receptor sensitivity (occupancy) during differentiation. The decrease of the IGF-II response with increased type II receptor with differentiation may on the other hand suggest that IGF-II in differentiated tissue no longer acts as a classical growth factor. These results tend to support the hypothesis of the switchover role of IGF-I and II during fetal and adult growth, however, confirmation of the precise role of IGF-I and II in biological growth may have to wait until further studies clarifying the significance of the increased IGF type II receptor in differentiated tissue are made.

Animals↗
PubMed2005

Changes in the freezing protocol for human zygotes alter embryonic development and pregnancy rates.

OBJECTIVE: To determine the effect of various freezing protocols on postthaw development and pregnancy rates resulting from transfer of human zygotes. DESIGN: Prospective study. SETTING: Tertiary care center. PATIENT(S): Couples undergoing assisted reproductive technology (ART) procedures who wished to have their excess zygotes cryopreserved. INTERVENTION(S): We cryopreserved zygotes with one of three protocols. MAIN OUTCOME MEASURE(S): Post-thaw survival and development of the zygotes as well as pregnancy rate after transfer of these zygotes. RESULT(S): A 3-minute hold time after seeding, followed by a final preplunging temperature of -180 degrees C, resulted in a clinical pregnancy rate of 28.6%. In contrast, a 15-minute postseed hold time and a -30 degrees C final chamber temperature resulted in a 37.3% clinical pregnancy rate. When we combined the protocols to provide a 15-minute postseed holding time and a -180 degrees C before plunging into liquid nitrogen, we achieved a 69.6% clinical pregnancy rate. CONCLUSION(S): By increasing the postseeding hold time and decreasing the temperature of the freezing chamber before plunging the zygotes into liquid nitrogen, significant improvements can be made in postthaw development and pregnancy rates.

Adult↗
PubMed2006

[Research progress on cell apoptosis during embryonic development of the zebrafish (Danio rerio)].

Cells have a built-in death program through the regulation of genes. Apoptosis (programmed cell death) plays a very important role in the development of the vertebrate embryo. The zebrafish (Danio rerio) has become a useful and ideal model in molecular developmental biology for in vivo study on the function of apoptosis in morphogenesis, sex differentiation and so on, and has already been garnering more and more attention. At present, much progress has been made regarding apoptotic pathways in zebrafish development. Caspases and other regulatory apoptotic genes have been cloned successfully. Moreover, transgenic zebrafish embryos have been induced to undergo apoptosis in vivo to study apoptotic signaling pathways and the abnormal morphological changes during embryogenesis. Transgenic zebrafish has provided a powerful tool for illuminating the interaction between regulatory apoptotic genes and vertebrate development.

Animals↗
PubMedDate not supplied

Targeted gene mutations define the roles of insulin and IGF-I receptors in mouse embryonic development.

Insulin-like growth factors (IGFs) and their receptors regulate embryonic and post-natal growth. Genetic evidence derived from targeted mouse mutants indicates that both the insulin receptor (IR) and IGF-I receptors (IGF-IRs) are required for mouse embryonic growth. However, the roles of IRs and IGF-IRs are functionally distinct, with IGF-IRs mediating both IGF-I and IGF-II actions, and IRs mediating IGF-II, rather than insulin, action. The combined interactions of IGF-IRs and IRs with IGF-I and IGF-II account for the entirety of the growth effects of these two ligands, and provide the molecular basis for IGFs-mediated intrauterine growth and differentiation. Genetic ablation experiments of insulin receptor substrate-1 (IRS-1) and -2 (IRS-2), two important molecules in the IR and IGF-IR signaling pathways, are also beginning to shed light onto the mechanisms accounting for the specificity of IR and IGF-IR signaling. IRS-1-deficient mice are growth retarded, while IRS-2-deficient mice develop diabetes, indicating that the two molecules play a more specific role than previously recognized in IGF-IR and IR signaling.

Animals↗
PubMed2004

Evidence that lactotrophs do not differentiate directly from somatotrophs during chick embryonic development.

It is generally accepted that, in mammals, lactotrophs differentiate from somatotrophs through an intermediate cell type, the mammosomatotroph. However, little information exists about mammosomatotrophs and their relationship with lactotroph development in non-mammalian vertebrates. We reported previously that corticosterone (CORT) can induce both somatotroph and lactotroph differentiation in cultures of chicken embryonic pituitary cells. Our current objectives were to determine the abundance of mammosomatotrophs during chicken pituitary development, to identify mammosomatotrophs during CORT induction of lactotrophs, and to explore whether lactotrophs induced by CORT are derived from somatotrophs. Cells that produced prolactin (PRL) only, growth hormone (GH) only or both hormones simultaneously were detected by three approaches - dual immunofluorescence, a combination of immunofluorescence and immunocytochemistry (ICC), and by ICC using combinations of antibodies to GH and PRL. Mammosomatotrophs were not detected between embryonic day (E) 16 and E20, even though lactotrophs increased from nearly absent to greater than 10% of all pituitary cells during this period. CORT induced more than 10% of all E13 pituitary cells to produce PRL, while the percentage of mammosomatotrophs remained at less than 1% of all cells. When cells from the cephalic and caudal lobes of the anterior pituitary were treated separately, CORT increased GH cells in cultures from the caudal lobe. No PRL cells were found in the caudal lobe. In the cephalic lobe, CORT increased lactotrophs, while GH cells were barely detected. In summary, mammosomatotrophs are rare during chicken pituitary development, and CORT does not induce lactotrophs from somatotrophs. These findings indicate that, unlike in mammals, lactotrophs do not differentiate from somatotrophs during chicken embryonic development.

Animals↗
PubMed2005

Gene expression profiling during the embryonic development of mouse brain using an oligonucleotide-based microarray system.

We analyzed gene expression profiles in embryonic day 12, 15, 18 and postnatal day 0 mouse brains by utilizing a GeneChip microarray. Significant differential expression was observed in 1413 of 12,422 (11.4%) represented on the chip. Then, 397 genes known to be related to neural development and functions were selected and analyzed in more detail. Clustering of the differentially expressed genes in terms of gene function and their temporal expression patterns indicated an aspect of the genetic foundation that underlies cellular events. Moreover, we identified a novel gene that encodes a putative protein kinase, Ebr kinase, which is differentially expressed in the developing brain.

Age Factors↗
PubMed2002

Dual roles for the Dab2 adaptor protein in embryonic development and kidney transport.

The Disabled-2 (Dab2) gene has been proposed to act as a tumor suppressor. Cell culture studies have implicated Dab2 in signal transduction by mitogens, TGFbeta and endocytosis of lipoprotein receptors. To identify in vivo functions of Dab2, targeted mutations were made in the mouse. In the absence of Dab2, embryos arrest prior to gastrulation with a phenotype reminiscent of those caused by deletion of some TGFbeta signal transduction molecules involved in Nodal signaling. Dab2 is expressed in the extra-embryonic visceral endoderm but not in the epiblast. Dab2 could be conditionally deleted from the embryo without affecting normal development, showing that Dab2 is required in the visceral endoderm but dispensable in the embryo proper. Conditionally mutant Dab2(-/-) mice are overtly normal, but have reduced clathrin-coated pits in kidney proximal tubule cells and excrete specific plasma proteins in the urine, consistent with reduced transport by a lipoprotein receptor, megalin/gp330, in the proximal tubule. This evidence indicates that Dab2 is pleiotropic and regulates both visceral endoderm function and lipoprotein receptor trafficking in vivo.

Adaptor Proteins, Signal Transducing↗
PubMed1995

Fixed patterns of bradycardia during late embryonic development in domestic fowl with C locus mutations.

A predictable late embryonic bradycardia (relative to normal White Leghorn chickens) has been documented in chicken strains with C locus mutations. The basis of the bradycardia remains unknown but clearly is related to a mutation at the C locus, which contains the structural gene for tyrosinase. When compared with the heart rate of normal White Leghorns (approximately 295-305 beats/min from day 8 to day 20 of incubation), ca/ca and other C locus mutants showed a 10-12% reduction in heart rate during the last 4 days of incubation. Embryonic mortality occurred in both mutant and normal strains at an equivalent rate (approximately 23%); a significant bradycardia (when compared with surviving embryos of the same strain) developed on the day before death in White Leghorn but not mutant strains. The bradycardia did not affect embryonic oxygen consumption (approximately 0.2 ml O2.egg-1.min-1 at day 14 and 0.4 ml O2.egg-1.min-1 at day 20), which showed only minor differences between strains that can be attributed to differences in embryonic mass on days 16-20.

Animals↗
PubMed2006

Expression of Six3 Opposite Strand (Six3OS) during mouse embryonic development.

Recently, sequence analyses have identified a large number of opposite strand transcripts in the vertebrate genome. Although the transcripts appear to be spliced and polyadenylated, many of them are predicted to represent noncoding RNAs. High levels of noncoding transcripts of the Six3 Opposite Strand (Six3OS) were recently identified in the embryonic and postnatal retina of the mouse. In this study, we expanded those initial expression analyses, elucidated in detail the developmental expression profile of mouse Six3OS in the brain and visual system, and compared it with that of Six3. Our results show that Six3OS expression overlaps extensively with that of Six3 and is not altered in Six3-null embryos.

Animals↗
PubMed1992

Changes in polyamines during embryonic development of Atlantic salmon, Salmo salar.

1. Polyamine composition of eggs and alevins from cultured and wild Atlantic salmon, Salmo salar, was compared during early development. 2. Eggs collected from wild stock had higher amounts of bound and free putrescine, spermidine and spermine than those collected from cultured stock. 3. Cadaverine was present in bound form in cultured and wild stocks before and after fertilization, respectively. However, free cadaverine was detected in both stocks throughout development. 4. Atlantic salmon embryos and alevins can synthesize polyamines which could be associated with their growth and development.

Animals↗