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R Wilk

Publications and source records attributed to R Wilk.

At least 19 recordsLinked to original sources
PubMed2001

Downregulation of Jun kinase signaling in the amnioserosa is essential for dorsal closure of the Drosophila embryo.

BACKGROUND: During Drosophila embryogenesis, Jun kinase (JNK) signaling has been shown to play a key role in regulating the morphogenetic process of dorsal closure, which also serves as a model for epithelial sheet fusion during wound repair. During dorsal closure the JNK signaling cascade in the dorsal-most (leading edge) cells of the epidermis activates the AP-1 transcription factor comprised of DJUN and DFOS that, in turn, upregulates the expression of the dpp gene. DPP is a secreted morphogen that signals lateral epidermal cells to elongate along the dorsoventral axis. The leading edge cells contact the peripheral cells of a monolayer extraembryonic epithelium, the amnioserosa, which lies on the dorsal side of the embryo. Focal complexes are present at the dorsal-most membrane of the leading edge cells, where they contact the amnioserosa. RESULTS: We show that the JNK signaling cascade is initially active in both the amnioserosa and the leading edge of the epidermis. JNK signaling is downregulated in the amnioserosa, but not in the leading edge, prior to dorsal closure. The subcellular localization of DFOS and DJUN is responsive to JNK signaling in the amnioserosa: JNK activation results in nuclear localization of DFOS and DJUN; the downregulation of JNK signaling results in the relocalization of DFOS and DJUN to the cytoplasm. The HINDSIGHT (HNT) Zn-finger protein and the PUCKERED (PUC) JNK phosphatase are essential for downregulation of the JNK cascade in the amnioserosa. Persistent JNK activity in the amnioserosa leads to defective focal complexes in the adjacent leading edge cells and to the failure of dorsal closure. CONCLUSIONS: Focal complexes are assembled at the boundary between high and low JNK activity. In the absence of focal complexes, miscommunication between the amnioserosa and the leading edge may lead to a premature "stop" signal that halts dorsalward migration of the leading edge. Spatial and temporal regulation of the JNK signaling cascade may be a general mechanism that controls tissue remodeling during morphogenesis and wound healing.

Amnion↗
PubMed2000

The hindsight gene is required for epithelial maintenance and differentiation of the tracheal system in Drosophila.

During animal development, morphogenesis of tissues and organs requires dynamic cell shape changes and movements that are accomplished without loss of epithelial integrity. Data from vertebrate and invertebrate systems have implicated several cell surface and cytoskeleton-associated molecules in the establishment and maintenance of epithelial architecture, but there has been little analysis of the genetic regulatory hierarchies that control epithelial morphogenesis in specific tissues. Here we show that the Drosophila Hindsight nuclear zinc-finger protein is required during tracheal morphogenesis for the maintenance of epithelial integrity and assembly of apical extracellular structures known as taenidia. In hindsight (hnt) mutants tracheal placodes form, invaginate, and undergo primary branching as well as early fusion events. Starting at midembryogenesis, however, the tracheal epithelium collapses or expands to give rise to sacs of tissue. While a subset of hnt mutant tracheal cells enters the apoptotic pathway, genetic suppression of apoptosis indicates that this is not the cause of the epithelial defects. Surviving hnt mutant tracheal cells retain cell-cell junctions and a normal subcellular distribution of apical markers such as Crumbs and DE-Cadherin. However, taenidia do not form on the lumenal surface of tracheal cells. While loss of epithelial integrity is a common feature of crumbs, stardust, and hnt mutants, defective assembly of taenidia is unique to hnt mutants. These data suggest that HNT is a tissue-specific factor that regulates maintenance of the tracheal epithelium as well as differentiation of taenidia.

Animals↗
PubMed1998

[Comparison of internal fixation methods for the symphysis in multi-directional dynamic gait simulation].

For the stabilization of the ruptured pubic symphysis, rigid forms of fixation such as plate osteosynthesis and flexible fixations such as wire loops or PDS banding have been recommended. All methods have only been tested by static unidirectional loading until failure of the system. By this experimental arrangement Ecke and Hofmann found comparable results for flexible and rigid methods of internal stabilization of the pubic symphysis. They preferred flexible methods to maintain mobility of the symphysis and to prevent symphyseal fusion. We tested dynamic compression plate osteosynthesis, reconstruction plate osteosynthesis, wire loops and PDS banding for internal fixation of injured pubic symphysis in a dynamic multidirectional experimental arrangement simulating gait conditions. The specimens were loaded with 85 N in vertical (y-) direction and 34 N in sagittal (z-) direction, which represent 50% of the forces acting at the pubic symphysis during walking and with a frequency of 1.5 Hz over 55,500 loads simulating the conditions over a 6-week mobilization period. Loading with 100% of the acting forces (corresponding to full weight bearing mobilization) led to early failure of the system. Our experimental analysis showed that neither wire loops nor PDS banding is able to stabilize the ruptured pubic symphysis, even immediately after fixation before loading. During the tests instability increased until failure of the system due to cutting of the bone or breaking of the wires or PDS banding. Success of plate osteosynthesis was dependent on the initial stability of the fixation. Overwinding of the screws, as in osteoporotic bone, lead to increasing loosening during repeated loading, whereas primary stable fixation of the screws was almost completely maintained during the test. In consequence, neither wire loops nor PDS banding should be used for stabilization of injured pubic symphysis if early mobilization with partial weight bearing is desired. Plate osteosynthesis (DC or reconstruction plate) tolerates early half weight bearing in patients with "open-book" injury only if safe screw fixation is guaranteed.

Biomechanical Phenomena↗
PubMed1998

Thyroid hormone inhibits aromatase activity in porcine thecal cells cultured alone and in coculture with granulosa cells.

We cultured porcine thecal and granulosa cells alone or in coculture to define whether thyroid hormone affects aromatase activity in porcine ovarian cells. Dispersed cells were cultured with 10(-9) M triiodothyronine (T3) for 24 hours. Testosterone (final concentration 10(-7) M) was added as aromatase substrate for granulosa cells (Gc) cultured alone. Thecal (Th) androgens serve as a substrate for estradiol secretion by Th cells cultured alone and in coculture with Gc. At the end of the preincubation time, the culture media was removed and replaced with fresh media containing 100 ng follicle stimulating hormone (FSH) or 10(-3) M 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP). After overnight incubation, the culture media was analyzed for estradiol production by radioimmunoassay (RIA). T3 inhibited basal, FSH-stimulated, and 8brcAMP-stimulated estradiol production in all culture conditions. T3 inhibited cAMP analogue 8-Br-cAMP and FSH-induced aromatase activity to a similar extent, thus suggesting that the inhibitory effect of T3 is downstream of cAMP formation. In the second part of the experiment a rabbit polyclonal antibody against human placental cytochrome P-450arom was used to confirm the effect of T3 on aromatase protein in Th and Gc. Pretreatment of Th and Gc with T3 markedly decreased immunostaining for aromatase in both types of cells, suggesting a direct effect of T3 on this enzyme.

Androgens↗
PubMed1996

UNC-40, a C. elegans homolog of DCC (Deleted in Colorectal Cancer), is required in motile cells responding to UNC-6 netrin cues.

UNC-6 netrin, a laminin-related protein secreted from neuroglia and neurons along the ventral midline, orients migrating cells and pioneering growth cones on the nematode epidermis. UNC-5, a cell surface protein expressed on motile cells and pioneer axons, orients movements away from UNC-6 sources. UNC-40, a homolog of the cell surface proteins DCC (Deleted in Colorectal Cancer) and neogenin, is also expressed on motile cells and pioneer neurons. UNC-40 acts cell autonomously to orient movement toward UNC-6 sources. For cells coexpressing UNC-5, it helps orient movement away from UNC-6 sources. Finally, UNC-40 helps determine the dorsoventral position of cells undergoing purely longitudinal migrations. Together with the recent report that DCC is a netrin receptor in vertebrates, our results suggest that UNC-40 is a component of UNC-6 receptors on motile cells.

Amino Acid Sequence↗
PubMed1996

trachealess encodes a bHLH-PAS protein that is an inducer of tracheal cell fates in Drosophila.

The embryonic tracheal system in Drosophila develops from placodes of precursor cells on the ectoderm. A transcription factor of the basic helix-loop-helix (bHLH)-PAS family, which is expressed in the nuclei of the tracheal cells throughout development, was identified. The protein shows the highest degree of homology to the Single-minded (Sim) protein. The transcript represents the previously identified trachealess (trh) locus, essential for tracheal development. Ectopic expression of trh leads to generation of extra tracheal pits and branches and to the expression of tracheal markers by patches of ectodermal cells. The expression of trh is consistent with a biphasic mode of transcriptional regulation. Expression is first induced by exogenous cues and is subsequently autoregulated. trh is also expressed and required in the posterior spiracles and the salivary gland ducts. The role of Trachealess in the formation of several tubular tissues in the embryo suggests that it may induce a general fate of branched tubular structures of epithelial origin.

Amino Acid Sequence↗
PubMed1996

[Biomechanics of the pubic symphysis. Which forces lead to mobility of the symphysis in physiological conditions?].

For estimation of the loads of fixation devices, physiological movements and acting forces at the symphysis pubis are examined. Experimentally, Walheim determined the movements of the pubic symphysis under physiological conditions (one-leg stance, walking): there is no effective mobility in the horizontal direction. During the one-leg stance he observed symphyseal mobility of up to 2.6 mm (Phi 1.2 mm) in the vertical direction and 1.3 mm (Phi 0.6 mm) in the sagittal direction. During walking he found symphyseal mobility of up to 2.2 mm (Phi 0.9 mm) in the vertical direction and 1.3 mm (Phi 0.6 mm) in the sagittal direction. Until now the forces leading to symphyseal mobility have not been estimated either experimentally or mathematically. In our experimental study we examined ten fresh cadaver anterior pelvic rings by means of a multidirectional force-mobility measurement. Maximal physiological movements, as determined by Walheim, were increasingly induced in ten equal steps, and the forces required were measured for every single step. Out of the resulting force-mobility curves for the ten cadaver specimens the mean force required (with standard deviation) was calculated for each of the ten measure points, separately for the vertical (y-) direction and sagittal (z-) direction. The values were graphically transferred, and a regression curve was created. This curve allows the acting force to be estimated for every movement of the pubic symphysis: During the one-leg stance the mean force to induce mean mobility is 169 N in vertical direction and 148 N in sagittal direction; for maximal mobility a force of 398 N in the vertical direction and 148 N in the sagittal direction is necessary. During walking, the force required to induce mean movement of the symphysis pubis is 120 N in the vertical direction and 68 N in the sagittal direction; for maximal mobility 333 N is required in the vertical direction and 136 N in the sagittal direction. For mobilization of patients with a symphyseal rupture (type Tile B 1) with partial weight-bearing, neutralization of the mean acting forces during the one-leg stance (169 N in the vertical direction, 68 N in the sagittal direction) must be achieved by an adequate fixation device. In cases where full weight bearing is desirable, such as in patients with limited compliance, stability can only be reached by neutralization of the maximal acting forces during walking (333 N in the vertical direction, 136 N in the sagittal direction).

Adult↗