Differential regulation of fast axonally transported proteins during the early response of rat retinal ganglion cells to axotomy.

We have found that the early response of axotomized rat retinal ganglion cells is characterized by the differential regulation of a number of fast axonally transported proteins. The abundance of 23 radiolabeled fast transported proteins was analyzed at 2 and 5 days after axotomy using two-dimensional gel electrophoresis. Corresponding changes in retinal GAP-43 mRNA were measured using northern analysis. Within 2 days of injury, > 40% of the transported proteins analyzed, including GAP-43, showed increased labeling above control levels. Approximately 13% of transported proteins decreased below control levels, whereas the remainder did not change. Five days after axotomy, only GAP-43 and another fast transported protein, C3, continued to sustain measurable increased labeling above control levels; all previously elevated proteins appeared to have been down-regulated by this time, which corresponds to the onset of cell death. These differential changes were accompanied by parallel increases in GAP-43 mRNA. These results suggest that the molecular changes within rat retinal ganglion cells are differentially regulated within two stages subsequent to damage, initial regenerative growth followed by cell death.

Two Drosophila regulatory genes, deformed and the Broad-Complex, share common functions in development of adult CNS, head, and salivary glands.

Deformed (Dfd), a homeotic selector gene required for segment identity in the head, and the Broad-Complex (BR-C), a steroid hormone-regulated locus required for metamorphosis of the epidermis and multiple internal tissues, are members of distinct genetic regulatory hierarchies. Their protein products contain DNA-binding domains (of the homeodomain and zinc-finger variety, respectively) and are believed to act by regulating the transcription of target genes. In this study we demonstrate that Dfd and BR-C mutants dying during metamorphosis share defects of CNS reorganization, ventral adult head development, and adult salivary gland morphogenesis. Specifically, the shared phenotypes are (i) failure to separate the subesophageal ganglion (SEG) from the thoracic ganglion (TG); (ii) structural and functional abnormalities of the proboscis and maxillary palps, innervated by the SEG; and (iii) failure of the adult salivary glands to extend into the thorax. Experiments performed with a conditional allele demonstrate that Dfd+ function during either larval life or metamorphosis is sufficient to rescue the SEG-TG separation phenotype. BR-C;Dfd double mutants show synergistic enhancement of the ventral head defects. This genetic interaction suggests that the segment identity and steroid hormone-sensitive regulatory hierarchies intersect during postembryonic development.

A genetic and developmental analysis of mutations in labial, a gene necessary for proper head formation in Drosophila melanogaster.

We have undertaken a developmental genetic analysis of labial (lab), the most proximal gene in the Antennapedia complex (ANT-C) of Drosophila melanogaster. The terminal phenotype of mutant embryos was examined in cuticle preparations, in thin sections, and by scanning electron microscopy. These preparations revealed a failure of head involution and the loss or disruption of several head structures, including the salivary glands and the H-piece and ventral arm of the cephalopharyngeal apparatus. Although these structures are presumed to derive from the gnathocephalic segments, we argue that the observed defects are likely to be a secondary consequence of a failure of head involution. A function for lab in the development of the adult head was inferred from the phenotype of animals bearing hypomorphic alleles and from clones of lab- tissue generated by mitotic recombination. Two aspects of the mutant phenotype were manifested. Ventrally, a deletion and/or disruption of tissue occurred in the maxillary palp and vibrissae regions. Dorsally, the posterior head appeared to be transformed to a thoracic-like identity. Mutations in lab, like those in the Deformed and proboscipedia loci of the ANT-C, reveal a homoeotic phenotype only in the adult stage of the life cycle.

Isolation, structure, and expression of labial, a homeotic gene of the Antennapedia Complex involved in Drosophila head development.

The labial (lab) gene of Drosophila melanogaster is necessary for the proper development of the embryonic (larval) and adult head. We have identified the lab transcription unit within the proximal portion of the Antennapedia Complex (ANT-C) by mapping the molecular lesions associated with chromosomally rearranged lab alleles. We present its molecular structure, nucleotide sequence, and temporal pattern of expression. In addition, using antibodies generated against a fusion protein, we show that in the embryo the lab protein is distributed in neural and epidermal cells of the procephalic lobe; in a discrete loop of the midgut; and in specific progenitor sensory cells of the clypeolabrum, thoracic segments, and tail region. The regions of lab expression in the developing cephalon represent nonsegmented domains that are anterior to and largely nonoverlapping with the domains of expression of the Deformed (Dfd) and proboscipedia (pb) genes, two other homeotic loci of the ANT-C that also function to direct the development of head structures. Furthermore, lab head expression is associated with the complex cellular movements of head involution, a process that not only is defective in lab embryos, but the failure of which appears to be largely responsible for the defects observed in mutant embryos. Finally, we suggest that lab head expression provides a molecular marker for an intercalary segment, an ancestral segment that has become morphologically indistinct during the evolution of the insect head.

A genetic and developmental analysis of mutations in the Deformed locus in Drosophila melanogaster.

Individuals expressing recessive mutations in the Deformed (Dfd) locus of Drosophila melanogaster were examined for embryonic and adult defects. Mutant embryos were examined in both scanning electron microscope and light microscope preparations. The adult Dfd recessive mutant phenotype was assessed in somatic clones and in survivors homozygous for hypomorphic alleles of the gene. The time of Dfd+ action was determined by studying a temperature conditional allele. Dfd+ is required in three embryonic cephalic segments to form a normal head. Mutant embryos of Dfd display defects in derivatives of the maxillary segment, of the mandibular segment, and of some more anterior segments. In the adult fly, defects are seen in the posterior aspect of the head when the gene is mutant. A transformation from head to thoracic-like tissue is seen dorsally and a deletion of structures is seen ventrally. Shift studies utilizing a temperature conditional allele have shown that the gene product is necessary during at least two periods of development, during embryonic segmentation and head involution and during the late larval and pupal stages. From these studies we conclude that Dfd is a homeotic gene necessary for proper specification of both the embryonic and the adult head.