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In the mouse embryonic maxilla, rudimentary tooth primordia have been identified, which can be mistaken for the first upper molar. In order to determine whether such a situation might exist in the lower jaw as well, tooth development was investigated in the mouse mandibular cheek region during ED 12.5-15.0. A combination of histology, morphometry and computer-aided 3D reconstructions demonstrated the existence of rudimentary dental structures, whose gradual appearance and regression was associated with the segmental progress of odontogenesis along the mesio-distal axis of the jaw: 1) At ED 12.5, the mesial segment (MS) was the most prominent part of the dental epithelial invagination. It included an asymmetrically budding dental lamina. The MS, although generally mistaken for the lower first molar (M1, primordium, regressed and did not finally participate in M1 cap formation. 2) At ED 13.5, a wide dental bud (called segment R2) appeared distally to the MS. Although the R2 segment transiently represented the predominant part of the dental epithelium at ED13.5, it participated only in the formation of the mesial end of the M1 cap. 3) The top of the R2 segment at ED13.5 was not the precursor of the enamel knot (EK), contrary to what has been assumed. 4) The central segment of the M1 cap as well as the EK developed later and distally to the R2 segment. 5) Time-space specific apoptosis correlated with the retardation in growth of the R2 segment as well as with strong regressive changes in the epithelium situated mesially to it. These highlight the need to reinterpret current molecular data on early M1 development in the mouse in order to correlate the expression of signalling molecules with specific morphogenetic events in the appropriate antemolar or molar segments of the embryonic mandible.
A study was undertaken to estimate gestational age, in terms of days from parturition, in medium and small size dogs by ultrasonographic examination. Serial ultrasonographic examinations were performed in four medium size pregnant bitches throughout two consecutive pregnancies and three small size pregnant bitches throughout one pregnancy, in order to determine the range of variation in the size of selected fetal structures throughout gestation. Formulae were derived to estimate the expected delivery date for both groups of bitches by measuring anatomical fetal structures, so that this method could be applied to a large number of different breeds. The determination of gestational age could be achieved with reasonable precision by selecting fetal structures. Prediction of parturition date was accurate to within one day by ultrasonographic measurement of the diameter of the gestational sac in early pregnancy and the biparietal diameter in late pregnancy in both small size breeds and medium size breeds.
15 cases of heart myxomas are presented. Their origin from remains of embryonal mesenchymal tissue is hypothesized. Predominant localization at the foramen ovale area, where remains of embryonal tissue can be found, and peculiarities of histological structure resembling embryonal mesenchyma may be proofs of the hypothesis. Microscopic structure of myxomas is homogeneous: branching cells of histiocytic origin amidst basophilic or eosinophilic stroma presented by myxoid tissue, capillaries and vessels of various gauges at different stages of functional activity (shut-down, compressed, dilated, filled with erythrocytes).
The intermesenteric plexus is an independent formation, it is connected with other plexuses in the abdominal and pelvic cavities and participates in innervation of organs. In the species investigated (mole, rat, cat, dog, man) connections between the intermesenteric and other vegetative plexuses in the abdominal and pelvic cavities vary according to their amount and complexity, they are most abundant and complex in man. In the latter the plexus is also mostly rich in neuro-fibrillar and neuro-cellular elements. In the intermesenteric plexus of the man and the animals studied there is a rather big part of vegetative ganglia, which can be considered as peripheral centers of the internal abdominal organs innervation.
Embryonic chick skeletal muscle has been shown to synthesize a distinct proteoglycan of large size with relatively large, highly 6-sulfated chondroitin sulfate glycosaminoglycans. Further analysis of these proteoglycans indicates that tryptic digestion gives rise to fragments with an average of two chondroitin sulfate chains per peptide. The skeletal muscle chondroitin sulfate proteoglycan also contains oligosaccharides whose characteristics suggest the presence of both O-linked and N-linked oligosaccharides. These characteristics include the average hydrodynamic size of the oligosaccharides as well as their localization. Approximately 10% of the putative O-linked oligosaccharides reside on the same tryptic fragments which contain the chondroitin sulfate chains, while the presumptive N-linked oligosaccharides appear to be present at sites distant from the chondroitin sulfate. Further support for this identification comes from radioisotopic labeling with [3H]mannose, which is incorporated exclusively into the putative N-linked oligosaccharides. Some of the O-linked oligosaccharides which are not in close apposition to the chondroitin sulfate seem to occur in clusters. The skeletal muscle chondroitin sulfate proteoglycan has the ability to interact in a link protein-stabilized fashion with hyaluronic acid. This ability as well as the estimated number of chondroitin sulfate chains per cluster and the estimated number of oligosaccharides per chondroitin sulfate chain have implications about the structure of the core protein of the skeletal muscle proteoglycan. The information presented is used to construct a model of these molecules; with this detailed model, attention can now be directed at other aspects of the skeletal muscle chondroitin sulfate proteoglycan, such as its role in myogenesis.
Three novel species-specific monoclonal antibodies 5.1.H, 8.7.D and 13.7.A raised against semi-purified detergent solubilized fractions of the Tera 1 embryonal carcinoma (EC) cell line are restricted in their in vitro distribution to undifferentiated human EC cell lines. Competition experiments have established that distinct antigenic specificities are seen by the 3 different monoclonal antibodies. All 3 antigens 5.1.H, 8.7.D and 13.7.A, defined by these monoclonal antibodies, undergo developmental regulation and cease to be expressed on Tera 2 clones 5 and 12 after retinoic-acid-induced differentiation and on LICR LON HT 39/7 cells after phorbol-ester-induced differentiation. These results taken together with the extremely limited in vivo tissue distribution of the defined antigens suggest that the 5.1.H, 8.7.D and 13.7.A monoclonal antibodies define distinct onco-foetal antigens.
An experiment was undertaken to determine which sensory structures of the mouse embryo inner ear developed from what portion of the mouse otocyst. Otocysts of gestation days 10, 11, 12, and 13 were divided by surgical dissection into six anatomical groups: dorsal, ventral, anterior, posterior, medial and lateral halves. They were organ cultured separately. After a period of ten days, the explanted tissues were harvested and processed histologically for microscopic analysis. The surgical control specimens fixed at the time of explantation were composed of undifferentiated ectodermal cells for tissues of gestation days 10, 11, and 12. Otocysts of gestation days 11, and 12 showed, during the course of their subsequent growth, that the three semicircular ducts and their associated cristae developed from the dorsal and lateral halves. Only the anterior and posterior canals and cristae originated from the medial portion. The posterior half gave rise to the posterior crista and the anterior half provided for the development of the anterior and lateral cristae. The cochlear duct and its sensory epithelium developed in all the anatomical groups except the dorsal half. The utricle developed in the dorsal section of the middle third of the otocyst, while the utricular macula developed in the anterior half of the same section of the otocyst. The saccule and its macula differentiated from the ventral section of the middle third of the anterior half.
Normal growth and development of the skeleton require the presence of viable, actively contracting skeletal muscle throughout the fetal period. A chick embryo model of midgestation chemical paralysis and secondary muscle atrophy was used to test the hypothesis that functioning muscle stimulates the growth of long bones by influencing the proliferation, differentiation, and hypertrophy of chondrocytes in cartilage of the epiphysis and growth plate. Paralysis did not alter the overall developmental stage of the long bone or the organization of the growth plate. Compared with controls, however, uptake of bromodeoxyuridine in the paralyzed chick was reduced by 27-55% in the chondroepiphysis and uppermost zone of the tibial growth plate, indicating reduced proliferation of chondrocytes. A specific reduction in the size of the proliferative zone and a reduced number of proliferating cells were also observed. By contrast, in the second, post-proliferative zone of the growth plate, the height of the zone was unchanged and its area was only slightly reduced compared with controls. Finally, median hypertrophic cell profile area, a measure of cell size, was not significantly affected by paralysis, although frequency analysis revealed modest numerical reductions in the population of the largest hypertrophic chondrocytes in the paralyzed group. These data suggest that the role of functioning fetal muscle in maintaining proper skeletal growth may be mediated primarily through specific stimulation of the recruitment or proliferation of immature chondrocytes, or of both.
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.
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cDNA clones originating from the engrailed gene of Drosophila have been isolated from recombinant phage libraries that were made using poly(A)+ RNA extracted from early embryos. The DNA sequence of one of these clones includes a homeo box, a 180 bp sequence present in several other Drosophila genes important in formation of body pattern during development. The homeo boxes found in the other Drosophila genes, as well as in cognate sequences from a wide range of segmented animals, including higher vertebrates, are highly conserved. By contrast, the homeo box within the engrailed gene diverges substantially and, unlike the other homeo boxes, is interrupted by an intervening sequence. The engrailed homeo box is located near the 3' end of a 1700 bp open reading frame. If translated, this sequence would produce a protein of unusual composition. We also show that a neighboring gene has a large region with strong homology to engrailed, and that it also contains a homeo box.
Cornified layers of newly hatched chick shank skin were solubilized in 8 M urea by reduction followed by S-carboxymethylations. On Sephadex gel chromatography the solubilized proteins were separated into two distinct protein fractions (protein A and protein B). SDS-gel electrophoresis showed that the smaller protein fraction (protein B) contained two molecules with molecular weights of 15,000 and 17,000. Protein B was resolved into several fractions, pI 4.5-5.2, by preparative isoelectric focusing in the presence of 6 M urea and 0.1% Nonidet P-40. The fractions all contained the two molecules (Mr 15,000 and 17,000). The amino acid compositions of these fractions and 2-dimensional electrophoresis of epidermal protein by the method of O'Farrel indicated that the two proteins are each heterogeneous with respect to charge for reasons of microheterogeneity in the amino acid composition and varying extent of phosphorylation. When chick embryonic skin was cultured in a chemically defined medium, hydrocortisone, which induces the synthesis of protein A and results in keratinization of epidermis (Sugimoto, M., Tajima, K., Kojima, A. and Endo, H. (1974) Dev. Biol. 39, 295-307), did not accelerate the accumulation of protein B in epidermis; the normal pattern of protein B was not formed in epidermis of cultured skin with or without the hormone. Actinomycin D did not inhibit the synthesis of protein B, suggesting that the mRNA for this protein has a long life.
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In a subset of papillary renal cell carcinomas a t(X;1)(p11;q21) chromosome translocation has repeatedly been reported. Positional cloning has demonstrated that, as a result of this translocation, the transcription factor TFE3 gene on the X-chromosome becomes fused to a novel gene, PRCC, on chromosome 1. Since as yet little is known about the function of PRCC, we sought to identify the mouse counterpart of the PRCC gene. Isolation and sequence analysis of a mouse Prcc cDNA revealed a high level of conservation between man and mouse, both at the nucleotide and protein level. As the human PRCC gene, the mouse Prcc gene is ubiquitously expressed. It shows low expression in all mouse fetal tissues examined. In addition, we identified a genomic cosmid clone containing the complete Prcc gene. The mouse Prcc gene consists of seven exons, all of which contain coding sequences. The small second exon, which was found to be located adjacent to the t(X;1) breakpoint in the human gene on chromosome 1, is also conserved between man and mouse. In mouse, Prcc is located on chromosome 3. These cDNA and genomic clones will be instrumental in the creation of mouse models for a further elucidation of the function of PRCC.
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