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1.
Oral Dis ; 22(5): 399-405, 2016 Jul.
Article in English | MEDLINE | ID: mdl-26866618

ABSTRACT

OBJECTIVES: GaAlAs lasers induce pulp mineralization by promoting reparative dentinogenesis. This study analyzed the expression of dentin matrix protein 1 (DMP1) and osteopontin in GaAlAs laser-irradiated rat molars, to examine the hypothesis that these proteins play a role in the laser-induced reparative dentinogenic process. MATERIALS AND METHODS: The mesial surfaces of the upper first molars of 8-week-old Wistar rats were irradiated with a pulsed GaAlAs laser. After 1-14 days, mRNA expression of DMP1 and osteopontin in the coronal pulp was analyzed using real-time PCR. DMP1, osteopontin, and heat shock protein 25 (HSP25) were immunolocalized at 1-21 days. RESULTS: The pulp exhibited a degenerative zone in its mesial portion on days 1-3, and progressive formation of reparative dentin lined with HSP25-immunoreactive odontoblast-like cells, from day 7 onwards. DMP1 and osteopontin mRNA expression were significantly upregulated on days 1-7 and 3-7, respectively. From day 7 onwards, DMP1 and osteopontin immunoreactivity colocalized along the boundary between the primary and reparative dentin. CONCLUSION: GaAlAs laser irradiation of rat molars induced upregulated DMP1 and osteopontin mRNA expression in the coronal pulp, followed by the formation of reparative dentin and the colocalization of DMP1 and osteopontin immunoreactivity at the site at which this tissue first appeared.


Subject(s)
Dentin, Secondary/metabolism , Dentin, Secondary/radiation effects , Extracellular Matrix Proteins/biosynthesis , Lasers, Semiconductor , Molar/radiation effects , Osteopontin/biosynthesis , Phosphoproteins/biosynthesis , Animals , Dental Pulp/cytology , Dental Pulp/physiology , Extracellular Matrix Proteins/radiation effects , HSP27 Heat-Shock Proteins/biosynthesis , Immunohistochemistry , Male , Molar/cytology , Molar/metabolism , Odontoblasts/metabolism , Odontoblasts/radiation effects , Osteopontin/radiation effects , Phosphoproteins/radiation effects , Rats , Rats, Wistar , Up-Regulation/radiation effects
2.
Int Endod J ; 48(6): 573-81, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25074651

ABSTRACT

AIM: To examine the temporospatial expression of dentine matrix protein 1 (DMP1; a noncollagenous protein involved in mineralized tissue formation), osteopontin (another noncollagenous protein detected during reparative dentinogenesis) and nestin (a marker of differentiating/differentiated odontoblasts), following direct pulp capping with calcium hydroxide in rat molars. METHODOLOGY: The maxillary first molars of 8-week-old Wistar rats had their pulps exposed and capped with calcium hydroxide. The pulp-capped teeth were collected from 6 h to 14 days postoperatively and processed for immunohistochemistry for DMP1, osteopontin and nestin. Cell proliferation was monitored using 5-bromo-2'-deoxyuridine (BrdU) labelling. RESULTS: The capped pulps initially exhibited superficial necrotic changes followed by the formation of new matrix and its mineralization. DMP1 immunoreactivity was observed in the matrix beneath the necrotic layer from 6 h onwards and present in the outer portion of the newly formed mineralized matrix from 7 days onwards. Osteopontin displayed a similar expression pattern, although it occupied a narrower area than DMP1 at 6 and 12 h. Nestin-immunoreactive cells appeared beneath the DMP1-immunoreactive area at 1 day, were distributed beneath the newly formed matrix at 5 days and exhibited odontoblast-like morphology by 14 days. BrdU-positive cells significantly increased at 2 and 3 days (P < 0.05) and then decreased. CONCLUSIONS: The deposition of DMP1 at exposed pulp sites preceded the appearance of nestin-immunoreactive cells, active cell proliferation and new matrix formation after pulp capping with calcium hydroxide in rat molars, suggesting that DMP1 acts as a trigger of pulp repair. The colocalization of DMP1 and osteopontin suggests that these two proteins play complementary roles.


Subject(s)
Dental Pulp Capping , Dental Pulp Necrosis/therapy , Extracellular Matrix Proteins/metabolism , Phosphoproteins/metabolism , Animals , Calcium Hydroxide , Cell Proliferation , Humans , Immunohistochemistry , Molar , Nestin/metabolism , Osteopontin/metabolism , Rats , Rats, Wistar
3.
Int Endod J ; 44(7): 669-75, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21375543

ABSTRACT

AIM: To investigate subcutaneous tissue reactions to methacrylate resin-based root canal sealers by immunohistochemical assessment of inflammatory/immunocompetent cell infiltration. METHODOLOGY: Silicone tubes containing freshly mixed Epiphany SE sealer, MetaSEAL, Super-Bond RC sealer, or a zinc oxide-eugenol sealer (Canals) were subcutaneously implanted into the backs of Wistar rats. Solid silicone rods implanted in different animals served as controls. After 7, 14 and 28 days, connective tissue surrounding the implants (n = 8, each) was processed for immunoperoxidase staining using OX6 (reactive to major histocompatibility complex class II molecules), ED1 (reactive to macrophages), and W3/13 (reactive primarily to neutrophils), and the number of positively stained cells within each field (1.2 × 0.8 mm) was enumerated. Statistical differences were analysed with Friedman's test and Scheffe's test (comparisons between test materials) or Mann-Whitney's U-test (test-control comparisons). RESULTS: Canals showed a significantly higher number of W3/13-positive cells (mostly neutrophils) than MetaSEAL at 28 days (P < 0.05). There were no significant differences in the numbers of OX6- or ED1-positive cells between each test material at any time point. Test-control comparisons revealed several significant differences for each antibody. This was most notable for ED1, where all the test materials at each time point, except for Epiphany SE at 28 days, showed significantly larger values than the corresponding controls. CONCLUSIONS: All the methacrylate resin-based sealers tested showed a similar level of inflammatory/immunocompetent cell infiltration. MetaSEAL induced less-intense neutrophil infiltration than Canals. Controls exhibited milder infiltration of inflammatory/immunocompetent cells compared with all the test materials.


Subject(s)
Methacrylates/pharmacology , Resin Cements/pharmacology , Root Canal Filling Materials/pharmacology , Subcutaneous Tissue/drug effects , Animals , Antibodies, Monoclonal , Antigens, CD , Antigens, Differentiation, Myelomonocytic , Boron Compounds/pharmacology , Cell Count , Connective Tissue/drug effects , Connective Tissue/pathology , Histocompatibility Antigens Class II/analysis , Immunoenzyme Techniques , Immunohistochemistry , Immunologic Factors/pharmacology , Inflammation Mediators/pharmacology , Leukocyte Count , Leukosialin , Lymphocytes/drug effects , Macrophages/drug effects , Male , Methylmethacrylates/pharmacology , Neutrophil Infiltration/drug effects , Neutrophils/drug effects , Rats , Rats, Wistar , Subcutaneous Tissue/pathology , Time Factors , Zinc Oxide-Eugenol Cement/pharmacology
4.
Dev Dyn ; 236(8): 2277-84, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17654720

ABSTRACT

We have performed in situ hybridization to study the expression of Wise in early chick embryos. Wise expression is first detectable in the ectoderm at posterior levels of late neurula. As development proceeds, Wise expression is seen in specific patterns in the ectoderm of the trunk region, pharyngeal arches, limb buds, and feather buds. In addition to these areas, particular cartilages such as the ones in the maxillary process and limbs start to express Wise at the late pharyngula stage, and the expression in these cartilages becomes stronger than that in epidermal components at later stages. Importantly, Wise is expressed in regions where other signaling molecules such as Wnt, Bmp, and Shh are known to function in morphogenesis and differentiation. Direct comparisons of the expression of Wise and these genes are also demonstrated.


Subject(s)
Gene Expression Regulation, Developmental , Intercellular Signaling Peptides and Proteins/genetics , Animals , Bone Morphogenetic Proteins , Cartilage/chemistry , Cartilage/embryology , Cell Differentiation , Chick Embryo , Ectoderm/chemistry , Hedgehog Proteins , Morphogenesis , Tissue Distribution , Wnt Proteins
5.
Bioessays ; 27(3): 331-8, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15714557

ABSTRACT

The jaw is one of the earliest innovations in vertebrate history. Several recent findings suggest a scenario for jaw evolution as a progression of changes in pharyngeal developmental mechanisms. The lamprey, an extant jawless vertebrate, constitutes a model for the pre-gnathostome ancestry. Comparing expression patterns of regulatory genes between the gnathostome and lamprey embryos may enable us to get a glimpse of the essential changes that were responsible for the evolution of the jaw. We hypothesize that a specific topographical change of inductive tissue interactions to be described here brought about the jaw as an evolutionary novelty.


Subject(s)
Biological Evolution , Jaw/anatomy & histology , Vertebrates/anatomy & histology , Animals , Body Patterning , Embryo, Nonmammalian/physiology , Fishes/anatomy & histology , Fishes/classification , Signal Transduction , Vertebrates/classification
6.
Int Endod J ; 35(4): 315-20, 2002 Apr.
Article in English | MEDLINE | ID: mdl-12059931

ABSTRACT

AIM: The aim of this study was to compare the sealing abilities of Fermin and Canseal with the more popular temporary coronal filling materials, Cavit and Caviton. METHODOLOGY: Standardized access cavities were prepared in 160 intact human permanent molar teeth. They were divided into five groups consisting of 32 samples. The teeth were restored using one of the temporary filling materials, namely: Fermin, Canseal at two powder to liquid ratios, Caviton and Cavit. Thermal cycling and/or load cycling were applied on the samples. Assessment of microleakage utilized methylene blue dye penetration. Grading of the microleakage pattern was from 1 to 3, with 3 providing the best seal. Results were analyzed using two-way anova and by Fisher's PLSD post hoc test (P < 0.05). RESULT: Microleakage along Fermin, Caviton and Cavit samples did not go beyond Leakage Grade 2. Dye penetration into these materials was noted. This was not observed in the two groups of Canseal tested. However, the two groups of Canseal exhibited total leakage notably after being subjected to thermal cycling. There was a statistically significant difference in the microleakage scores obtained between the materials and conditions tested (P < 0.0001). CONCLUSION: Fermin was found to exhibit the best seal amongst the four materials tested followed by Caviton, and Cavit. Thermal cycling influenced the seal of certain types of temporary filling materials more than load cycling.


Subject(s)
Dental Cements/chemistry , Dental Leakage/classification , Root Canal Filling Materials/chemistry , Analysis of Variance , Calcium Sulfate/chemistry , Coloring Agents , Drug Combinations , Humans , Methylene Blue , Polyvinyls/chemistry , Root Canal Obturation , Root Canal Preparation , Statistics as Topic , Thermodynamics , Vinyl Compounds/chemistry , Weight-Bearing , Zinc Oxide/chemistry , Zinc Sulfate/chemistry
7.
Philos Trans R Soc Lond B Biol Sci ; 356(1414): 1615-32, 2001 Oct 29.
Article in English | MEDLINE | ID: mdl-11604127

ABSTRACT

Evolution of the vertebrate jaw has been reviewed and discussed based on the developmental pattern of the Japanese marine lamprey, Lampetra japonica. Though it never forms a jointed jaw apparatus, the L. japonica embryo exhibits the typical embryonic structure as well as the conserved regulatory gene expression patterns of vertebrates. The lamprey therefore shares the phylotype of vertebrates, the conserved embryonic pattern that appears at pharyngula stage, rather than representing an intermediate evolutionary state. Both gnathostomes and lampreys exhibit a tripartite configuration of the rostral-most crest-derived ectomesenchyme, each part occupying an anatomically equivalent site. Differentiated oral structure becomes apparent in post-pharyngula development. Due to the solid nasohypophyseal plate, the post-optic ectomesenchyme of the lamprey fails to grow rostromedially to form the medial nasal septum as in gnathostomes, but forms the upper lip instead. The gnathostome jaw may thus have arisen through a process of ontogenetic repatterning, in which a heterotopic shift of mesenchyme-epithelial relationships would have been involved. Further identification of shifts in tissue interaction and expression of regulatory genes are necessary to describe the evolution of the jaw fully from the standpoint of evolutionary developmental biology.


Subject(s)
Biological Evolution , Jaw/physiology , Lampreys/embryology , Animals , Cartilage/embryology , Embryo, Nonmammalian , Head/anatomy & histology , Head/embryology , Jaw/embryology , Lampreys/physiology , Mesoderm , Mouth/embryology , Mouth/physiology , Neural Crest/physiology
8.
Genesis ; 30(2): 51-8, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11416863

ABSTRACT

The Thyroid transcription factor-1 (TTF-1) gene belongs to the Nkx-2.1 subfamily and encodes a transcription factor containing an NK-2-type homeodomain. In our study, we isolated and characterized cDNA clones for the TTF-1/Nkx-2.1 orthologue (LjTTF-1) from the agnathan vertebrate Lampetra japonica. Spatial and temporal expression patterns assessed by in situ hybridization revealed the expression of LjTTF-1 in the anterior nerve cord and anteroventral region of the pharynx. The neural expression was subsequently restricted to the ventral diencephalon. The pharyngeal expression, on the other hand, extended posteriorly to the fourth pharyngeal-pouch level and was finally localized in the endostyle anlage. In the differentiated endostyle of ammocoete larvae, the expression of LjTTF-1 was chiefly detected in type 2a, 2b, and 2c cells, which develop adjacent to glandular cells. These expression patterns of LjTTF-1 support the idea that this gene family plays an important role in the development of the rostral brain and endostyle equivalent organs. Furthermore, histological comparisons between TTF-1/Nkx-2.1 expression in the endostyles of ammocoetes and ascidians suggested the possibility that the organogenetic architecture of the endostyle is conserved among chordates.


Subject(s)
Gene Expression , Homeodomain Proteins/genetics , Lampreys/genetics , Nuclear Proteins/genetics , Thyroid Gland/metabolism , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Biological Evolution , Brain/cytology , Brain/embryology , Cloning, Molecular , Female , Homeodomain Proteins/chemistry , Humans , Lampreys/embryology , Male , Sequence Homology, Amino Acid , Thyroid Gland/embryology , Thyroid Nuclear Factor 1
9.
J Exp Zool ; 291(1): 68-84, 2001 Apr 15.
Article in English | MEDLINE | ID: mdl-11335917

ABSTRACT

Agnathan cognates of vertebrate homeobox genes, Emx and Dlx, were isolated from embryonic cDNA of a Japanese marine lamprey, Lampetra japonica. Analyses of amino acid sequences indicated that the Dlx cognate was closely related to the common ancestor of gnathostome Dlx1 and Dlx6 groups and termed LjDlx1/6. Southern blot analyses could not rule out the possibility that L. japonica possesses more than one paralog for both LjDlx1/6 and LjEmx, the lamprey cognate of Emx. Expression of LjDlx1/6 was regulated spatially as well as developmentally, and its transcripts were mainly found in the craniofacial and pharyngeal mesenchyme and in the forebrain. The expression pattern of LjEmx changed dramatically during embryogenesis; expression was seen initially in the entire neural tube and mesoderm, which were secondarily downregulated, and secondarily in cranial nerve ganglia and in the craniofacial mesenchyme. No specific expression of LjEmx was seen in the telencephalon. Comparisons of Dlx and Otx gene expression patterns suggested a shared neuromeric pattern of the vertebrate brain. Absence of Emx expression implied that the patterning of the lamprey telencephalon is not based on the tripartite plan that has been presumed in gnathostomes. Expression domains of LjDlx1/6 in the upper lip and of LjEmx in the craniofacial mesenchyme were peculiar features that have not been known in gnathostomes. Such differences in expression pattern may underlie distinct morphogenetic pathway of the mandibular arch between the agnathans and gnathostomes.


Subject(s)
Conserved Sequence , Evolution, Molecular , Gene Expression Regulation, Developmental , Genes, Homeobox , Head/embryology , Homeodomain Proteins/genetics , Lampreys/genetics , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Female , Homeodomain Proteins/biosynthesis , Homeodomain Proteins/isolation & purification , Humans , Lampreys/embryology , Male , Mice , Molecular Sequence Data , Sequence Alignment , Sequence Homology, Amino Acid , Transcription Factors/biosynthesis , Transcription Factors/isolation & purification , Zebrafish
10.
Dev Biol ; 228(1): 73-85, 2000 Dec 01.
Article in English | MEDLINE | ID: mdl-11087627

ABSTRACT

The most rostral cephalic crest cells in the chick embryo first populate ubiquitously in the rostroventral head. Before the influx of crest cells, the ventral head ectoderm expresses Fgf8 in two domains that correspond to the future mandibular arch. Bmp4 is expressed rostral and caudal to these domains. The rostral part of the Bmp4 domain develops into the rostral end of the maxillary process that corresponds to the transition between the maxillomandibular and premandibular regions. Thus, the distribution patterns of FGF8 and BMP4 appear to foreshadow the maxillomandibular region in the head ectoderm. In the ectomesenchyme of the pharyngula embryo, expression patterns of some homeobox genes overlap the distribution of their upstream growth factors. Dlx1 and Barx1, the targets of FGF8, are expressed in the mandibular ectomesenchyme, and Msx1, the target of BMP4, in its distal regions. Ectopic applications of FGF8 lead to shifted expression of the target genes as well as repatterning of the craniofacial primordia and of the trigeminal nerve branches. Focal injection of a lipophilic dye, DiI, showed that this shift was at least in part due to the posterior transformation of the original premandibular ectomesenchyme into the mandible, caused by the changed distribution of FGF8 that defines the mandibular region. We conclude that FGF8 in the early ectoderm defines the maxillomandibular region of the prepharyngula embryo, through epithelial-mesenchymal interactions and subsequent upregulation of homeobox genes in the local mesenchyme. BMP4 in the ventral ectoderm appears to limit the anterior expression of Fgf8. Ectopic application of BMP4 consistently diminished part of the mandibular arch.


Subject(s)
Ectoderm/metabolism , Epithelium/metabolism , Fibroblast Growth Factors/metabolism , Head/embryology , Mesoderm/metabolism , Animals , Body Patterning/drug effects , Bone Morphogenetic Protein 4 , Bone Morphogenetic Proteins/administration & dosage , Bone Morphogenetic Proteins/metabolism , Bone Morphogenetic Proteins/pharmacology , Cell Differentiation/drug effects , Chick Embryo , Ectoderm/cytology , Ectoderm/drug effects , Ectoderm/ultrastructure , Epithelium/drug effects , Epithelium/ultrastructure , Fibroblast Growth Factor 8 , Fibroblast Growth Factors/administration & dosage , Fibroblast Growth Factors/pharmacology , Fluorescent Antibody Technique , Gene Expression Regulation, Developmental/drug effects , Genes, Homeobox/genetics , In Situ Hybridization , Mesoderm/cytology , Mesoderm/drug effects , Mesoderm/ultrastructure , Microscopy, Electron, Scanning , RNA, Messenger/genetics , RNA, Messenger/metabolism , Signal Transduction/drug effects
11.
Dev Biol ; 223(2): 399-410, 2000 Jul 15.
Article in English | MEDLINE | ID: mdl-10882524

ABSTRACT

Among the transcription factor gene families, Pax genes play important and unique roles in morphological patterning of animal body plans. Of these, Group I Pax genes (Pax1 and Pax9) are expressed in the endodermal pharyngeal pouches in many groups of deuterostomes, and vertebrates seem to have acquired more extensive expression domains in embryos. To understand the evolution of Pax1/Pax9-related genes in basal groups of vertebrates, their cognates were isolated from the Japanese marine lamprey, Lampetra japonica. RT-PCR of larval lamprey cDNA yielded two different fragments containing vertebrate Pax1- and Pax9-like paired domains. The Pax9 orthologue was isolated and named LjPax9. Whole-mount in situ hybridization revealed that this gene was expressed in endodermal pharyngeal pouches, mesenchyme of the velum (the oral pumping apparatus) and the hyoid arch, and the nasohypophysial plate, but not in the somitic mesoderm of the lamprey embryo. These expression patterns could be regarded as a link between the basal chordates and the gnathostomes and are consistent with the phylogenetic position of the lamprey. Especially, the appearance of neural crest seemed to be the basis of velar expression. Homology of the velum and the jaw is also discussed based on the LjPax9 expression in the first pharyngeal pouch and in the velar mesenchyme. We conclude that Pax9 genes have sequentially expanded into new expression domains through evolution as more complicated body plans emerged.


Subject(s)
Biological Evolution , Body Patterning , Lampreys/embryology , Lampreys/genetics , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Gene Library , Larva , Molecular Sequence Data , Multigene Family , Neural Crest/embryology , Paired Box Transcription Factors , Pharynx/embryology , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Tissue Distribution
12.
Dev Growth Differ ; 40(4): 423-9, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9727356

ABSTRACT

In the early chick embryo, the dorsal ventral (DV) boundary organizes the apical ectodermal ridge (AER) structure in the limb bud field. Here it is reported that Engrailed-1 (En-1), a homolog of the Drosophila segment polarity gene engrailed expressed in the ventral limb ectoderm, participates in AER formation at the DV boundary of the limb bud. Restricted ectopic expression of En-1 in the dorsal side of the limb bud by transplantation of En-1-overexpressing ectoderm induces ectopic AER at the boundary of En-1-positive and -negative cells. The results suggest that En-1 is involved in AER formation at the DV boundary of the limb bud.


Subject(s)
Ectoderm/metabolism , Ectoderm/transplantation , Homeodomain Proteins/physiology , Limb Buds/metabolism , Neural Crest/metabolism , Animals , Cell Differentiation/genetics , Cell Differentiation/physiology , Chick Embryo , Chimera/genetics , Ectoderm/cytology , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Developmental/physiology , Genetic Vectors/genetics , Homeodomain Proteins/analysis , Homeodomain Proteins/genetics , In Situ Hybridization , Limb Buds/cytology , Limb Buds/embryology , Neural Crest/cytology , Transfection
13.
Neurosci Res ; 27(3): 211-7, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9129179

ABSTRACT

The retinotectal projection map is organized in a precise retinotopic order, so that the temporo-nasal axis of the retina corresponds to the rostro-caudal axis of the tectum. en-1 and en-2, homologues of the Drosophila segment polarity gene engrailed, are expressed in a gradient along the rostro-caudal axis of the tectal anlage, and are suggested to confer caudal characteristics as the results of transplantation and ectopic engrailed (en) expression. Recently the ligands for Eph type receptor tyrosine kinases have been shown to be expressed strongly at the caudal tectum and play a role in retinotectal map formation by repulsing the temporal retinal fibers. Using the system of replication competent retroviral vector, en-2 RCAS (A/B), we misexpressed en-2 on the tectum. Elf-1 or RAGS was induced at the ectopic En-2 sites. The present results have shown that En-2 can regulate expression of both Elf-1 and RAGS. This suggests that the cells which express en at the early stage of tectum development acquire positional specificity as 'caudal' tectum, and these cells may later express the ligands for Eph type receptor tyrosine kinases. Therefore the temporal retinal fibers which have the receptors are repelled when they meet the ligands on the tectum.


Subject(s)
Gene Expression Regulation, Developmental , Protein-Tyrosine Kinases/metabolism , Retina/metabolism , Animals , Base Sequence , Chick Embryo , Genes, Homeobox , Ligands , Molecular Sequence Data
14.
Jpn J Cancer Res ; 86(6): 540-5, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7622418

ABSTRACT

Parthenogenesis has been suggested to be tightly coupled with development of ovarian teratomas. Indeed, ovarian tumors developed in c-mos-deficient female mice, which are characterized by the parthenogenetic activation of oocytes. The tumors appeared at a frequency of 30% between 4 and 8 months of age, and did not develop in younger or older mice. Most of the tumors were benign and consisted of multi-focal cysts most notably with mature ectodermal components, but also with mesodermal and endodermal components. One among 17 tumors observed consisted of extra-embryonic tissues alone, and two bore malignant components with metastasis to peritoneal organs. The results strongly suggest the involvement of c-mos mutations in human germ cell tumors.


Subject(s)
Genes, mos , Ovarian Neoplasms/genetics , Teratoma/genetics , Age Factors , Animals , Female , Heterozygote , Homozygote , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Ovarian Cysts/genetics , Ovarian Cysts/pathology , Ovarian Neoplasms/pathology , Teratoma/pathology
15.
Oncogene ; 9(9): 2433-40, 1994 Sep.
Article in English | MEDLINE | ID: mdl-8058305

ABSTRACT

Fyn is a member of the Src family of tyrosine kinases which are thought to play important roles in cell to cell interactions during morphogenesis. The developmental profile of Fyn expression was examined using mutant mice in which lacZ gene was introduced into this locus. The expression was characteristic in the neural system. Though at low levels, it was detected in the headfold at embryonic day (E) 7.5 and in the luminal surface of neuroectoderm along the entire neural groove at E8.5. The expression appeared regional in rhombomeres at E8.5 and E9.5. Consistent expression was also found at a low level in the notochord. The expression was high in later stages of the neural tube which consists of three layers; it was in the marginal layer but not in the germinal layer. High expression was also found in developing dorsal root filaments of neural crest origin. Non-expression in dividing neuroepithelial cells and expression in developing neural fibers appeared ubiquitous features of Fyn expression throughout the entire brain.


Subject(s)
Nervous System/embryology , Proto-Oncogene Proteins/analysis , Animals , Base Sequence , Female , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Nervous System/metabolism , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins c-fyn , beta-Galactosidase/analysis
16.
Nature ; 366(6457): 742-5, 1993.
Article in English | MEDLINE | ID: mdl-8264796

ABSTRACT

Non-receptor-type tyrosine kinases of the Src family, such as Src, Yes and Fyn, are strongly expressed in the brain and have been suggested to have an important function in the central nervous system. We generated Fyn-deficient mice by inserting the beta-galactosidase gene (lacZ) into the fyn gene. The homozygous Fyn-mutant neonates from homozygous Fyn-deficient parents died because of a suckling problem. Neonates were, however, able to suckle milk normally when the homozygous mother's mammary glands had been activated by suckling of a heterozygous or wild-type pup. In these homozygous pups, the modified glomerular complex of the olfactory bulb, which had been suggested to play a role in perceiving pheromones, was abnormal in shape and reduced in size, and the hippocampal cell-layer was undulated. These results suggest that Fyn may be involved in the initial step of instinctive suckling behaviour in neonates.


Subject(s)
Protein-Tyrosine Kinases/physiology , Proto-Oncogene Proteins/physiology , Sucking Behavior/physiology , Animals , Animals, Suckling , Heterozygote , Homozygote , Mammary Glands, Animal/physiology , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Mutagenesis , Olfactory Bulb/physiology , Proto-Oncogene Proteins c-fyn
17.
Oncogene ; 8(12): 3343-51, 1993 Dec.
Article in English | MEDLINE | ID: mdl-8247536

ABSTRACT

Mutant mice in which beta-galactosidase gene (lacZ) was inserted into fyn locus were generated by homologous recombination in embryonic stem cells to examine the Fyn expression in the central nervous system. In adult brain, intensive beta-galactosidase activity was observed in olfactory bulb, cerebellum and hippocampus of the limbic system; the subcellular distribution of the activity was apparent not only in cell body but also in neural processes, and homozygous mutant mice live-born displayed an anatomical abnormality in the neural cell layer of the hippocampal formation. In spinal cord it was specifically expressed in dorsal horn, and in brain stem it was more characteristic in the sensory pathway, suggesting roles of Fyn in the sensory nervous network. In the white matter area, it was intense at postnatal day 10 but not detectable in adult, suggesting Fyn's role in myelinization.


Subject(s)
Brain Chemistry , Lac Operon/genetics , Proto-Oncogene Proteins/analysis , Proto-Oncogene Proteins/genetics , Proto-Oncogenes/genetics , Animals , Base Sequence , Blotting, Western , Cerebellum/chemistry , Cerebellum/cytology , Cerebellum/embryology , Cerebral Cortex/chemistry , Cerebral Cortex/cytology , Cerebral Cortex/embryology , Embryo, Mammalian/chemistry , Embryo, Mammalian/cytology , Gene Expression/genetics , Heterozygote , Hippocampus/chemistry , Hippocampus/cytology , Hippocampus/embryology , Homozygote , In Situ Hybridization, Fluorescence , In Vitro Techniques , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Inbred ICR , Mice, Mutant Strains , Molecular Sequence Data , Mutation , Nervous System/chemistry , Nervous System/cytology , Nervous System/embryology , Olfactory Bulb/chemistry , Olfactory Bulb/cytology , Olfactory Bulb/embryology , Proto-Oncogene Proteins/physiology , Proto-Oncogene Proteins c-fyn , Spinal Cord/chemistry , Spinal Cord/cytology , Spinal Cord/embryology , beta-Galactosidase/genetics , beta-Galactosidase/metabolism , beta-Galactosidase/physiology
18.
Proc Natl Acad Sci U S A ; 87(24): 9918-22, 1990 Dec.
Article in English | MEDLINE | ID: mdl-2263643

ABSTRACT

In attempting to produce a mutant mouse with embryonic stem cells, the critical step is the efficient isolation of homologous recombinants; the frequency of the homologous recombination is usually low and the potency of the cells to differentiate into germ cells is unstable in culture. Here, we report an efficacious method for such isolation in which the diphtheria toxin A-fragment gene is used to negatively select nonhomologous recombinants. In contrast to the use of the herpes simplex virus thymidine kinase gene, the selection can be made singly by the neomycin analog G418 without using a drug such as ganciclovir, a nucleoside analog. At the c-fyn locus, the diphtheria-toxin negative selection enriched the recombinants about 10-fold, and half of the cells integrating with the neomycin phosphotransferase gene were homologous recombinants.


Subject(s)
Diphtheria Toxin/genetics , Proto-Oncogene Proteins/genetics , Proto-Oncogenes , Animals , Base Sequence , Blotting, Southern , Cell Line , DNA/genetics , DNA/isolation & purification , Embryo, Mammalian , Female , Gene Library , Genetic Vectors , Hematopoietic Stem Cells , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Molecular Sequence Data , Oligonucleotide Probes , Protein-Tyrosine Kinases/genetics , Proto-Oncogene Proteins c-fyn , Restriction Mapping , Selection, Genetic
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