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Plasma patterns of prolactin, progesterone, and estradiol during early pregnancy in aging rats: relation to embryonic development.

Regularly cyclic, middle-aged female rats exhibit a decreased incidence of fertility, and those females that are fertile produce smaller litters. This decreased litter size is directly related to a reduced number of normal blastocysts available for implantation. Recent evidence indicates that embryonic abnormalities in middle-aged rats become apparent as early as Day 2 of pregnancy. Inasmuch as the semicircadian secretion of prolactin (PRL) is essential for the rescue of corpora lutea during early gestation and luteal production of progesterone (P) and estradiol (E2) in sufficient quantities is obligatory for embryonic development and implantation, the present study examined the profiles of plasma PRL, P, and E2 during the first 3 days of pregnancy in both young and middle-aged rats and assessed the embryonic development in these same animals. Regularly cyclic, middle-aged (9-11 mo) and young (4-5 mo) rats were cannulated via the right jugular vein on Diestrus Day 2 and mated with fertile males on proestrus. The next morning, sperm in the vaginal lavage confirmed mating, and that day was designated Day 1 of pregnancy. Beginning at 1400 h on Day 1 and continuing to 2400 h on Day 2, serial blood samples were taken at 2-h intervals for PRL assay. In the first experiment, samples were also collected at 8-h intervals during Days 1-3 for measurement of plasma P.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging

Galactosyltransferase activities during embryonic development of chick neural tissue.

Galactosyltransferase specific activities in embryonic chick retina, optic tectum, and telencephalon were found to decline during embryonic development. Incorporation of galactose from nucleotide sugar into exogenously added glycoprotein acceptor was measured in the presence of excess of glycoprotein acceptor. This ensured that the specific activity measurements were reflections of tru enzyme specific activity rather than availability of acceptor. Moreover, we have shown that the decline in specific activity is not due to degradation of the nucleotide sugar, UDP-galactose, under our in vitro assay conditions. Enzymatic specific activity declined sharply with embryonic age for all tissues tested. This decline was not affected by the presence of 5-bromodeoxyuridine during in vitro culture of embryonic chick neural retina above that caused by the culturing alone. Galactosyltransferase activity was not found to be associated with the plasma membrane fraction from homogenized tissue but rather with the microsomal fraction. Thus, the changes in galactosyltransferase specific activity detected here do not reflect changes at the cell surface.

Animals

The importance of cytoplasm in early embryonic development.

PURPOSE: A mouse model was used to evaluate the consequences of partial cytoplasm removal, which mimics the consequence of human early embryo fragmentation, at the one-cell stage on embryonic development to the morula and blastocyst stages. RESULTS: Opening the zona pellucida at the zygote stage has no detrimental effect on the number of differentiated or total cells at morula/blastocyst stage. However, reduced cytoplasm significantly affects the number of inner cell mass cells, 21.7 (10% cytoplasm removed) vs 27.6 (control) and 15.1 (20% cytoplasm removed) vs 24.7 (control). CONCLUSIONS: The contribution of cytoplasm of maternal origin is critical to early embryonic development.

Animals

Effects of cyproheptadine on the rat yolk sac membrane and embryonic development in vitro.

Electron-microscopic examinations of rat embryonic yolk sacs treated in vitro with 1.5 X 10(-5) M cyproheptadine showed proliferation of the lysosomal structures; other organelles remained unaffected, and also overall yolk-sac growth and vascularization were comparable to non-treated samples. Radioactive measurements with 125I-labelled albumin showed that yolk sacs and embryos of the cyproheptadine-treated group incorporated less radioactivity than the controls. Embryos inside the yolk sacs, treated either for 24 or 48 h, were severely retarded in growth and differentiation (approximately 50% of the controls). It is suggested that the specific action of cyproheptadine on yolk-sac lysosomal structures, combined with reduced macromolecular transport, is the cause of inhibited embryonic development.

Animals

Glycoprotein synthesis and embryonic development.

One of the most striking morphogenetic events during embryonic development is gastrulation, a process that leads to formation of the primitive gut. Using sea urchin embryos, we have studied the synthesis and function of glycoproteins during gastrulation. These studies have revealed that at least three processes are induced prior to gastrulation: de novo synthesis of dolichol; phosphorylation of dolichol by dolichol kinase, which may catalyze the final step in the de novo pathway; and initiation of the synthesis of N-linked glycoproteins. Whether or not activation of the glycosylation process results merely because of the production of dolichyl monophosphate or because, in addition, proteins containing glycosylatable-Asn-X-Ser/Thr-sequences are first translated just prior to gastrulation, is currently being investigated.

Animals

Transfer of IgA from albumen into the yolk sac during embryonic development in the chicken.

In the chicken, maternal antibodies are transferred into the egg and subsequently transported into the developing embryo. IgG is the primary immunoglobulin isotype of the egg yolk, while IgM and IgA are mainly found in the albumen. However, considerable amounts of IgM and IgA of unknown origin are found one day prior to hatching in the yolk sac. These antibodies are not synthesized de novo by the embryo proper, thus pointing to a transfer from the albumen into the egg yolk during development. To further address this question, 125I labelled chicken IgA was injected into the albumen of freshly laid eggs. On day 21 of embryonic development, 125I-IgA was found in the yolk sac content. On average (n = 6) 36.2 +/- 7.2% of the injected radioactivity was recovered from this compartment and shown to be associated with IgA. Comparison of total IgA in the albumen of freshly laid eggs with the amounts of IgA in yolk sac content showed similar results with a 44% transfer rate. An increase of the IgA concentration in the yolk sac was first detectable between days 14 and 16 of embryonic development. These data clearly show that IgA is transferred from the albumen into the yolk sac, most likely by a transport across the yolk sac membrane.

Animals

PAX-genes expression during human embryonic development, a preliminary report.

PAX-genes encode important transcriptional factors during embryogenesis. They are also involved in human diseases, Waardenburg syndrome, Aniridia and tumors. We report in the present paper a preliminary in situ hybridization study of PAX3-, PAX5- and PAX6-gene expression during human embryonic development. PAX3-gene is expressed in the neural groove before closure, and in the closed neural tube. Afterwards, its expression is observed in the mesencephalon, the rhombencephalon and the spinal cord. PAX5-gene expression is restricted to the mesencephalon-rhombencephalon boundary and the spinal cord. PAX6-gene is expressed early in the neural tube, just after its closure. Afterwards, its expression is observed in the forebrain, the rhombencephalon, the somites and the spinal cord. These patterns of expression are observed early during human embryonic development and are specific in time and space. This preliminary report shows the feasibility of in situ hybridization methodology for studying the expression of developmental genes during the early stages of human embryogenesis. It opens the way to study the pathogenesis of polymalformative syndromes and tumorigenesis.

Chromosome Mapping

[The embryonic development of the ostiomeatal complex from 8 to 40 weeks].

The embryonic development of the ostiomeatal complex from 8-40 weeks was studied under the light microscope. From our observation, the uncinate process was visible on the laterosuperior portion of the inferior turbinate at 8 week's gestation. By 12 weeks, the ethmoid bulla was first identifiable on the lateral wall of the middle meatus. The primordial ethmoidal infundibulum and primordial maxillary sinus were seen developing lateral to the uncinate process in the middle meatus. The air cells of the middle turbinate may be normal development of the ethmoidal labyrinth. Congenital nasal septum deviation may cause malformation of the ostiomeatal complex.

Embryonic and Fetal Development

Massive loss of mid- and hindbrain neurons during embryonic development of homozygous lurcher mice.

The mouse neurological mutant lurcher (Lc) results from a semidominant mutation. Heterozygous Lc/+ mice are viable but ataxic because Lc/+ Purkinje cells die by apoptosis within the first 3 weeks of life. Lc/Lc mice die shortly after birth. To aid in understanding the function of the lurcher gene product, we have examined the embryonic development of homozygous lurcher animals. The ratio of +/+:Lc/+:Lc/Lc animals did not deviate significantly from the expected 1:2:1. Homozygous lurcher mice at P0 were found to be normal under gross morphological examination. However, these mice weighed less, lacked milk in their stomach, and died within the first day of life. No resorbed embryos were found at embryonic day (E) 17.5, indicating that all homozygous lurchers survived until birth. Histological examination of P0 animals revealed that in homozygous lurcher mice the patterning of the brain is normal but that there has been a massive loss of hindbrain neurons during embryonic development. A particularly conspicuous consequence of the Lc/Lc genotype at birth is the complete absence of large neurons comprising the trigeminal motor nucleus. These neurons arise normally and are maintained until E15.5. However, beginning at E15.5 large numbers of pyknotic cells are evident in the trigeminal motor nucleus, suggesting that these cells die coincident with their terminal differentiation in the developing hindbrain. Because the trigeminal motor nucleus controls muscles required for suckling, these results suggest an explanation for the neonatal death of homozygous Lc animals. These data demonstrate that the severe and dose-dependent developmental consequences of lurcher gene action result from degeneration of distinct neuronal populations on maturation in the developing CNS.

Animals

Differential expression of inhibin subunits and follistatin, but not of activin receptor type II, during early murine embryonic development.

Activins are known to be potentially important regulators of early developmental processes in amphibians, birds, and mammalians. In this study we report the expression of the inhibin subunits, including those that make up activin, the activin-binding protein follistatin, and activin receptor type II in several in vitro systems that model early murine embryonic development, namely embryonic stem (ES) cells, embryonal carcinoma (EC) cells, and their differentiated derivatives. In addition, we examine the expression pattern of these factors in different stages of the mouse embryo itself. Expression of inhibin alpha and beta A subunits is restricted to certain differentiated cell types, while beta B subunits are expressed in both differentiated and undifferentiated cells. Our results further indicate a change in the expression pattern of inhibin subunits during early development from beta B at the blastocyst stage largely to beta A in postgastrulation embryos. This is similar to the expression pattern at equivalent stages of Xenopus and chick development. Expression of the activin-binding protein follistatin is altered by the induction of differentiation of P19 EC and ES cells by several factors, including retinoic acid. In contrast to the inhibin subunits and follistatin, activin receptor levels are not influenced by differentiation in these cell types. The results of this study demonstrate that the inhibin subunits and follistatin, but not the activin receptor type II, are differentially expressed during early murine development and suggest that the different forms of activin/inhibin are involved in the regulation of different developmental processes.

Activin Receptors

The embryonic development of the human ethmoid labyrinth from 8-40 weeks.

The embryonic development of the human ethmoid labyrinth was studied in 24 fetal heads aged between 8 and 40 weeks of gestation under light microscopy. The uncinate process was identifiable at 8 weeks of gestation on the laterosuperior portion of the inferior turbinate; however, at this stage of development, the ethmoid bulla was not apparent. The ethmoid bulla developed on the lateral wall of the middle meatus by 12 weeks of gestation. By 14 weeks, the primordial ethmoid infundibulum and primordial maxillary sinus were seen developing between the uncinate process and the ethmoid bulla. It was obvious that the anterior and middle ethmoid cells developed from the ethmoid bulla. By 22 weeks of gestation, the first cell of the anterior ethmoid group was evident in the anterior-inferior portion of the ethmoid bulla. By 23 weeks of gestation, the first cell of the middle ethmoid group was visible in the superior portion of the ethmoid bulla. Pneumatization of the middle turbinate occurred as part of normal development of the ethmoid labyrinth. By 32 weeks of gestation, the ostium for the development of the middle turbinate cell was seen in the superior-interior portion of the middle turbinate. These observations provide new insight into the development of the ethmoid labyrinth and have important implications for the understanding of normal anatomy and developmental variants of the ethmoid labyrinth.

Embryonic and Fetal Development

Cardiac specific expression of the green fluorescent protein during early murine embryonic development.

We demonstrate the establishment of transgenic mice, where the expression of the green fluorescent protein (GFP) is under control of the human cardiac alpha-actin promoter. These mice display cardiac specific GFP expression already during early embryonic development. Prominent GFP fluorescence was observed at the earliest stage of the murine heart anlage (E8). Cardiomyocytes of different developmental stages proved GFP positive, but the intensity varied between cells. We further show that contractions of single GFP positive cardiomyocytes can be monitored within the intact embryo. At later stages of embryonic development, the skeletal musculature was also GFP positive, in line with the known expression pattern of cardiac alpha-actin. The tissue specific labeling of organs is a powerful new tool for embryological as well as functional investigations in vivo.

Actins

Transcription of HLA-G transgenes commences shortly after implantation during embryonic development in mice.

We studied the pattern of transcription of a human HLA-G transgene in mice using polymerase chain reaction (PCR) techniques. Transcription of the HLA-G transgene commenced in cells derived from embryos as soon as 48 hr after implantation of embryos in the uterine wall and continued for at least a further 48 hr during embryonic development. HLA-G transcripts were also present in RNA extracted from thymus, spleen and liver of adult HLA-G transgenic mice, although transcripts were not detected in RNA from any other tissues except testes of male transgenic mice. These results demonstrate that the restricted pattern of HLA-G transcription in embryo-derived trophoblast cells during the first trimester of human pregnancy is reproducible in mice. This suggests that transcription factors required for a highly regulated pattern of gene expression during embryonic development are present in murine trophoblast cells and provide a means to investigate the factors and study the consequences of HLA-G expression during development of the embryo.

Animals

Changes in the expression of alpha-fodrin during embryonic development of Xenopus laevis.

Fodrin (nonerythroid spectrin) and its associated proteins have been previously implicated in the establishment of specialized membrane-cytoskeletal domains in differentiating cells. Using antiserum which is monospecific for the alpha-subunit of fodrin, we demonstrate that alpha-fodrin is present in oocytes and adult tissues of Xenopus laevis. Analyses of the de novo synthesis of alpha-fodrin during embryonic development reveal that alpha-fodrin is synthesized in oocytes, but not during early development. To investigate the level of control of alpha-fodrin expression, we isolated two cDNA clones for oocyte alpha-fodrin. The oocyte cDNA clones were identified as encoding portions of alpha-fodrin based on DNA sequence analysis and on the comparison of the predicted amino acid sequence of the cDNAs with the known sequence of human erythrocyte alpha-spectrin. The Xenopus alpha-fodrin cDNAs hybridize to a transcript of approximately 9 kb on RNA blots, and probably to a single gene type on genomic DNA blots. Both RNA blot analyses and S1 nuclease protection assays with the Xenopus alpha-fodrin cDNAs demonstrate that the observed decline in the de novo synthesis of alpha-fodrin polypeptides is controlled by a dramatic decrease in the abundance of alpha-fodrin transcripts after fertilization. In contrast, levels of actin transcripts do not decrease during this period. Inasmuch as steady-state levels of alpha-fodrin transcripts rise by the neurula stage of development, these results suggest that the synthesis of alpha-fodrin polypeptides during embryonic development of Xenopus is regulated, rather than constitutive, and that the primary level of control is the steady-state abundance of mRNA.

Amino Acid Sequence

An abnormal pattern of embryonic development during early pregnancy in aging rats.

There is recent evidence that a decline in fertility and litter size precedes the cessation of regular estrous cyclicity in middle-aged female rats. This decline in litter size is related to a decrease in the number of normal blastocysts that are present on Day 5 of gestation, immediately prior to implantation. Thus, the pattern of embryonic development during the first 5 days of pregnancy may be altered in middle-aged rats, resulting in fewer implanting embryos and smaller litter sizes. The present study examined the ovulation rates, fertilization rates, and the patterns of embryonic development in regularly cyclic, young and middle-aged females during the first 5 days of pregnancy. Examination of the numbers of ovulated ova revealed that the ovulation rate was significantly reduced in 12- to 14-mo-old females (13 mo; 9.0 +/- 1.0/rat), but not in 9- to 11-mo-old females (10 mo; 12.2 +/- 0.8/rat), as compared to that in young animals (12.8 +/- 1.0/rat). However, there was no decrease in fertilization rate in either the 10-mo or 13-mo group. While the total numbers of embryos present on Days 2-5 were similar among all 3 groups, embryos from 10-mo females displayed a delayed pattern of development and an increased incidence of morphological abnormalities. These changes in embryo development were even more pronounced in the 13-mo group. By Day 5 of pregnancy there was a significant reduction in normal blastocysts in 10-mo (7.3 +/- 1.2/rat) and 13-mo (6.0 +/- 1.6/rat) rats, as compared to young females (10.6 +/- 0.9/rat).(ABSTRACT TRUNCATED AT 250 WORDS)

Aging

A stereological study of the different cell populations in chicken testes treated with follicle-stimulating hormone during embryonic development.

Quantitative morphological methods were used to analyse the histomorphometric changes and variations in the number and size of cells from diverse cellular populations in testes of newly hatched chicks treated with follicle stimulating hormone (FSH) during embryonic development. The tissue was fixed and embedded in Epon and sections were morphometrically measured under light microscopy, using point counting for volume densities and the Floderus equation to determine numerical density. The average volume of the individual cell was determined by dividing the volume density by the numerical density. Results indicate that FSH administration causes an increase in the number of Sertoli cells and spermatogonia, as well as enlargement of the individual Sertoli cells leading consequently to an increase in the diameter and volume density of the testicular seminiferous tubules. Results also reveal an increase in the volume density of the interstitial cords of the Leydig cells, this expansion is due to the enlargement of the individual Leydig cells and not to an increase in their number, which remains constant. We conclude that testes of chick embryos are able to respond to FSH treatment, as revealed by the changes in the number and size of the cells conforming the diverse cellular populations of the testis. FSH treatment during embryonic development induces histomorphometric changes in both the interstitial tissue and seminiferous tubules, accelerating their growth and differentiation.

Animals

Antibiotic dipping studies in relation to uptake, embryonic development and Arizona hinshawii recoveries from turkey hatching eggs.

Antibiotic dipping with gentamicin sulfate by means of the temperature-differential method was effective in reducing but not completely eliminating Arizona hinshawii (7:1, 7, 8) from artificially infected turkey hatching eggs. Embryonic development was well maintained. The antibiotic dip solution intake was variable from egg to egg. Removal of cuticle by means of either 0.2 N HCl or a 10% disodium salt of ethylene diamine tetraacetic acid enhanced egg weight gains and maintained good embryonic development. Complete elimination of A. hinshawii was achieved in one experiment out of four. Correlation studies between egg weight gains and gentamicin concentrations of the contents of the turkey hatching eggs indicated a statistically significant relationship between these two parameters.

Animals

Selection for B cells with productive IgL gene rearrangements occurs in the bursa of Fabricius during chicken embryonic development.

The vast majority of immunoglobulin-expressing mature chicken B lymphocytes contain one functionally rearranged and one unrearranged allele of the immunoglobulin light chain (IgL) gene. Therefore, nearly all IgL V-J rearrangements present in mature chickens are in-frame. In contrast, the Ig genes of mature mammalian B cells contain a high proportion of out-of-frame V-J joints. To investigate the basis for this difference, gene rearrangement at the chicken IgL locus was characterized during embryonic development and in mature B-cell lines. Joining of the single functional variable (VL) segment with the single joining (JL) segment occurs in cells in multiple tissues during a transient period of chicken embryogenesis. Only one-third of the V-J joints cloned from days 10-12 of development are in-frame. An increasing proportion of in-frame V-J joints is observed within the bursa of Fabricius at successively later stages of development. Our data suggest that the bursa of Fabricius serves during embryonic development as a site of selective amplification of cells that have undergone productive V-J joining, such that nearly all V-J joints present in postembryonic B cells are in-frame. The high frequency of rearranged alleles joined in-frame that is found in posthatching bursal cells and mature B-cell lines appears to result from a low frequency with which cells undergo IgL rearrangement at both alleles, rather than from an increase in the precision of V-J joining in avian species.

Alleles