A Bonafede | PubMed Health

PubMed2000

Tissue-specific distribution and variation of the channel-forming protein ductin during development of Drosophila melanogaster.

Ductins represent membrane channel proteins which are supposed to form both proton channels in V-ATPases and connexon channels in gap junctions. In order to localize and characterize these proteins in different tissues of Drosophila, we applied indirect immunofluorescence microscopy and immunoblots, using antisera prepared against Drosophila ductin and against Nephrops ductin. Previously, these antisera have been shown to recognize, in ovarian follicles and young embryos of Drosophila, the ductin monomer of 16 kDa and a putative dimer of 29 kDa. Moreover, both anti-ductin sera label antigens in plasma membranes and in the cytoplasm and block, when microinjected, cell-cell communication via gap junctions. In the present study, comparing several embryonic, larval and adult tissues, the anti-ductin sera were found to recognize antigens with various locations in cells of the midgut, the salivary gland, the nervous system, the muscles and the epidermis. For example, in midgut cells, antigens were labeled mainly in apical plasma membranes and in the apical part of the cytoplasm, while in salivary-gland cells, labeling was found throughout the plasma membranes and the cytoplasm. We conclude that putative gap junctions were revealed in the salivary gland, the nervous system and the epidermis, while plasma membrane-associated putative V-ATPases were detected in the midgut, the salivary gland and the muscles. Moreover, V-ATPases associated with cytoplasmic vesicles were found in almost every tissue. On immunoblots of homogenates from various tissues, the anti-ductin sera specifically labeled bands of 16, 21 and 29 kDa. When comparing these bands using peptide mapping with V8 protease, we found that they represent closely related proteins. Therefore, either different ductins or modifications of a single ductin appear to be present in different cellular regions, cell types and developmental stages of Drosophila.

Adenosine Triphosphatases↗

PubMed1999

SF/HGF is a mediator between limb patterning and muscle development.

Scatter factor/hepatocyte growth factor (SF/HGF) is known to be involved in the detachment of myogenic precursor cells from the lateral dermomyotomes and their subsequent migration into the newly formed limb buds. As yet, however, nothing has been known about the role of the persistent expression of SF/HGF in the limb bud mesenchyme during later stages of limb bud development. To test for a potential role of SF/HGF in early limb muscle patterning, we examined the regulation of SF/HGF expression in the limb bud as well as the influence of SF/HGF on direction control of myogenic precursor cells in limb bud mesenchyme. We demonstrate that SF/HGF expression is controlled by signals involved in limb bud patterning. In the absence of an apical ectodermal ridge (AER), no expression of SF/HGF in the limb bud is observed. However, FGF-2 application can rescue SF/HGF expression. Excision of the zone of polarizing activity (ZPA) results in ectopic and enhanced SF/HGF expression in the posterior limb bud mesenchyme. We could identify BMP-2 as a potential inhibitor of SF/HGF expression in the posterior limb bud mesenchyme. We further demonstrate that ZPA excision results in a shift of Pax-3-positive cells towards the posterior limb bud mesenchyme, indicating a role of the ZPA in positioning of the premuscle masses. Moreover, we present evidence that, in the limb bud mesenchyme, SF/HGF increases the motility of myogenic precursor cells and has a role in maintaining their undifferentiated state during migration. We present a model for a crucial role of SF/HGF during migration and early patterning of muscle precursor cells in the vertebrate limb.

Animals↗