The role of apoptosis in early embryonic development of the adenohypophysis in rats.

BACKGROUND: Apoptosis is involved in fundamental processes of life, like embryonic development, tissue homeostasis, or immune defense. Defects in apoptosis cause or contribute to developmental malformation, cancer, and degenerative disorders. METHODS: The developing adenohypophysis area of rat fetuses was studied at the embryonic stage 13.5 (gestational day) for apoptotic and proliferative cell activities using histological serial sections. RESULTS: A high cell proliferation rate was observed throughout the adenohypophysis. In contrast, apoptotic cells visualized by evidence of active caspase-3, were detected only in the basal epithelial cones as an introducing event for fusion and closure of the pharyngeal roof. CONCLUSION: We can clearly show an increasing number of apoptotic events only at the basic fusion sides of the adenohypophysis as well as in the opening region of this organ. Apoptotic destruction of epithelial cells at the basal cones of the adenohypophysis begins even before differentiation of the adenohypophyseal cells and their contact with the neurohypophysis. In early stages of development, thus, apoptotic activity of the adenohypophysis is restricted to the basal areas mentioned. In our test animals, the adenohypophysis develops after closure of the anterior neuroporus.

Induction and prevention of cleft lip, alveolus and palate and neural tube defects with special consideration of B vitamins and the methylation cycle.

Previous studies on the development of cleft lip, alveolus, palate, and velum and neural tube defects have revealed several shared multifactorial causes. Both anomalies emerge at different times during embryonic development and are not associated with each other unless there is a genetic component to the etiology. Vitamin deficiency disorders are one of several factors contributing to the etiology of these anomalies.Vitamins B6, folic acid and B12 play an essential role in the methylation cycle. A lack of or deficiency in these vitamins thus has severe consequences for the organism, especially the unborn child. Due to its short half-life, vitamin B6 is particularly important for undisturbed embryogenesis and should be taken along with folic acid as a periconceptional supplement to prevent embryonic deformities. This paper is intended to provide the orthodontist (as a member of the interdisciplinary cleft team) with an overview of the issues and etiological significance of vitamin B deficiencies as possible inducers of these embryopathies. This may encourage comprehensive counselling, particularly of parents of children born with deformities who wish to have more children.

In vivo study of apoptosis as a creative agent of embryonic development of the primary nasal duct in rats.

INTRODUCTION: The first embryonic part of the nasal cavity is the primary nasal duct, beginning with the olfactory placode and ending with the oronasal membrane. Aim of this study was to investigate the cellular processes (apoptosis, proliferation) being responsible for development and opening of the primary nasal duct. MATERIAL AND METHODS: In this study developmental processes in at least three regions of the primary nasal duct (opening, middle, end) were examined by sectioning 38 rat fetuses on day 13.5 after conception. Apoptotic cells were detected by active caspase-3 antibodies and proliferating cells were examined by Ki-67 antibodies. RESULTS: Multiple apoptotic events were diagnosed on the basis and proliferative cells on the top of this duct. CONCLUSION: Apoptosis and proliferation play an important role in the process of opening the bottom of the primary nasal duct and for development of the nasal septum, philtrum as well as the primary palate. Mesenchymal proliferation seems to play a minor role in the process of opening the primary nasal duct.

Apoptosis as a creative agent of embryonic development of bucca, mentum and nasolacrimal duct. An in vivo study in rats.

INTRODUCTION: For embryonal facial development several fusion processes between different facial prominences are necessary. If fusion fails to appear, various facial clefts may occur, known as median (e.g. lower median cleft lip), oblique (e.g. open nasolacrimal duct) or lateral facial clefts (macrostomia, lateral cleft). MATERIAL AND METHODS: The development of 3 different facial regions (bucca, mentum, and nasolacrimal duct) was examined in rats using serial histological sections on day 13.5 after conception. Common procedures were used (staining for active caspase-3 and for Ki-67) for histological assessment about the role of apoptotic and proliferative processes in the fusion zones of buccal, mental and nasolacrimal areas. RESULTS: Multiple apoptotic events were detected in epithelial cells of the respective regions, the proliferative centers were located in the mesenchymal surroundings of fusion zones. CONCLUSION: A substantial precondition for fusion of facial prominences are proliferative and apoptotic processes in epithelial and mesenchymal cells. Apoptosis contributes to the development of bucca, mentum and the nasolacrimal duct. Absence of apoptoses may be responsible for facial clefts.

Suppression of the TIG3 tumor suppressor gene in human ovarian carcinomas is mediated via mitogen-activated kinase-dependent and -independent mechanisms.

The TIG3 gene is a retinoic acid inducible class II tumor suppressor gene downregulated in several human tumors and malignant cell lines. Diminished TIG3 expression correlates with decreased differentiation whereas forced expression of TIG3 suppresses oncogenic signaling pathways and subsequently induces differentiation or apoptosis in tumor cells. Analysis of TIG3 mRNA expression in a large set of cDNA pools derived from matched tumor and normal human tissues showed a significant downregulation of TIG3 in 29% of the cDNA samples obtained from ovarian carcinomas. Using in situ hybridization, we demonstrated expression of TIG3 in the epithelial lining of 7 normal ovaries but loss of TIG3 expression in 15/19 of human ovarian carcinoma tissues. In SKOV-3, CAOV-3 and ES-2 ovarian carcinoma cell lines, downregulation of TIG3 mRNA was reversible and dependent on an activated MEK-ERK signaling pathway. Re-expression of TIG3 mRNA in these cells upon specific interference with the MEK-pathway was correlated with growth inhibition of the cells. In OVCAR-3 and A27/80 ovarian carcinoma cells, TIG3 suppression is MEK-ERK independent, but expression could be reconstituted upon interferon gamma (IFNgamma) induction. Overexpression of TIG3 in A27/80 ovarian carcinoma cells significantly impaired cell growth and despite increased mRNA levels, TIG3 protein was hardly detectable. These results suggest that TIG3 is negatively regulated by an activated MEK-ERK signaling pathway. Further mechanisms must interfere with TIG3 expression that are independent of MEK and partially include interferon-responsive components.

BRUCE, a giant E2/E3 ubiquitin ligase and inhibitor of apoptosis protein of the trans-Golgi network, is required for normal placenta development and mouse survival.

BRUCE is a highly conserved 528-kDa peripheral membrane protein of the trans-Golgi network. Owing to the presence of an N-terminal single baculovirus inhibitor repeat, BRUCE functions as an inhibitor of apoptosis protein and blocks apoptosis when overexpressed. In addition, due to the presence of a C-terminal ubiquitin-conjugating domain, BRUCE can covalently attach ubiquitin to substrates. Here we report the generation and characterization of BRUCE-deficient mice. Complete inactivation of the BRUCE gene resulted in perinatal lethality and growth retardation discernible after embryonic day 14. The growth defect is linked to impaired placental development and may be caused by insufficient oxygen and nutrient transfer across the placenta. Chorioallantoic placentation initiated normally, but the mutant placenta showed an impaired maturation of the labyrinth layer and a significant reduction of the spongiotrophoblast. No evidence for an elevated apoptosis rate was detectable in embryonic and extraembryonic tissues and in knockout fibroblasts. Thus, although BRUCE is broadly expressed in embryonic, extraembryonic, and adult mouse tissues, this bifunctional protein might play a unique role in normal trophoblast differentiation and embryonic survival.

Mapping of epitopes on the fiber knobs of human adenovirus serotypes 8 and 15.

In order to obtain information on the linear antigenic epitopes on fiber knobs of adenovirus serotypes 8 (Ad8) and 15 (Ad15) of subgenus D, the binding of polyclonal virus- and fiber-specific rabbit antibodies to overlapping peptides covering the fiber knob was studied. The main antigenic epitopes of the fiber knob of Ad8 (FK8) were represented by the peptides P4 [amino acids (aa) 213-227], P6 (aa 233-247), P11 (aa 283-297), P13 (aa 303-317) and P15 (aa 316-325); the peptides P1 (aa 183-197), P8 (aa 253-267), P10 (aa 273-287) and P23 (aa 340-349) were moderately reactive. The peptides P4, P6, P11, P13 and P15 span the beta strands C, D, G and H, parts of the CD and DG loops and the complete GH loop of the fiber knob. The main epitopes of the fiber knob of Ad15 (FK15) were represented by peptides P5 (aa 198-212), P10 (aa 223-237), P12 (aa 233-247), P13 (aa 238-252), P26 (aa 303-317), P29 (aa 318-332) and P32 (aa 333-347), spanning the beta strands B, D, G, H and I, partly strand C, the CD loop, parts of the AB and DG loops, the GH and HI loops and the N-terminal part of the IJ loop. The amino acid sequence alignment showed that the location of the linear FK8 and FK15 epitopes was found to be overlapping to a major extent. Two serotype-specific epitopes were determined on FK15, represented by P10 and P13.