Octopod, a homeotic mutation of the moth Manduca sexta, influences the fate of identifiable pattern elements within the CNS

Octopod (Octo) is a mutation of the moth Manduca sexta, which results in the homeotic transformation of the ventral surface of the first (A1) and less often the second (A2) abdominal segments in the anterior direction. The extent of the transformation ranges from a slight deformation of the ventral cuticle, up to the formation of miniature thoracic legs on A1. The extent of the transformation is always less within A2 as compared to A1. A genetic analysis revealed that Octo is an autosomal mutation which shows incomplete dominance. The effect of this mutation on the central nervous system (CNS) was assessed by examining the distribution and fate of the postembryonic neuroblasts in the segmental ganglia of Octo larvae. In each of the thoracic ganglia of wild-type larvae, there is a set of 45-47 neuroblasts; a reduced but homologous array of 24 and 10 neuroblasts are found in A1 and A2, respectively. Ganglion A1 of Octo larvae had 1 to 6 supernumerary neuroblasts, and 20% of the A2 ganglia showed a single ectopic neuroblast. The supernumerary neuroblasts corresponded to identifiable neuroblasts normally found in more anterior ganglia. The Octo mutation also influenced the mitotic activity of stem cells normally present in A1. In this case, the neuroblasts generated a lineage of cells that were typical of a thoracic location rather than A1. These data demonstrate that homeotic mutations can influence the fate of identifiable pattern elements within the CNS of an insect.     

Radiosensitive mutants of Drosophila. VI. The effect of ultraviolet rays and methylmethane sulfonate on the viability and incidence of chromosome aberrations in the somatic cells of mutant mus(2)201G1

The mus(2)201G1 mutation determining high sensitivity to UV-rays and methyl methansulfonate (MMS) has been studied. The larvae of Drosophila of different age were treated with UV-rays and MMS. Lethality of organisms during the larvae and the pupa stages of the development, as well as the frequency of spontaneous and induced chromosome aberrations were registered. The mus(2)201G1 mutation was shown to determine high lethality of Drosophila during larvae and pupa stages as well as a high frequency of spontaneous and induced chromosome aberrations. The conclusion was made that chromosome aberrations are not the single reason for the death of the mutant flies after mutagenic treatment and that the function of the mus(2)201G1 gene is necessary for divided and undivided cells.     

Segregation of the somatic and germ-cell pathways of development in Drosophila

Two groups of lethal mutations that are expressed only in somatic cells were isolated using the gonosomic selective method. The mutations tend to be organized in clusters along the sex chromosome. Analysis of mutation expression allowed to characterize the degree of changes in genetic diversity of somatic and germ lines in development; it is minimal in the III instar larvae and prepupae and maximal in the I and II instar larvae and pupae.     

Precocious metamorphosis in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori

Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs). JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several "moltinism" mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and characterization of the gene responsible for the dimolting (mod) mutant that undergoes precocious metamorphosis with fewer larval-larval molts. We show that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued by topical application of a JH analog. We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. We also demonstrated that CYP15C1 is specifically expressed in the corpus allatum, an endocrine organ that synthesizes and secretes JHs. Furthermore, a biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. Precocious metamorphosis of mod larvae was rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. Our data therefore reveal that CYP15C1 is the gene responsible for the mod mutation and is essential for JH biosynthesis. Remarkably, precocious larval-pupal transition in mod larvae does not occur in the first or second instar, suggesting that authentic epoxidized JHs are not essential in very young larvae of B. mori. Our identification of a JH-deficient mutant in this model insect will lead to a greater understanding of the molecular basis of the hormonal control of development and metamorphosis.     

A natural flightless mutation in the ladybird, Harmonia axyridis

A homozygous flightless strain, obtained by selection from a laboratory population of Harmonia axyridis Pallas (Coleoptera, Coccinellidae), has been reared under controlled conditions since 1982 after sampling in China. The adults of this flightless strain have normal elytra and wings but drop almost vertically when they attempt to fly. The origin of this mutation, whether laboratory or natural, is discussed. As the mutation does not affect the fitness of the beetles, this flightless strain can be used in biological control. Releasing flightless adults rather than larvae should provide the potential for more continuous control of aphids by both larvae and adults.     

Chromosome aberrations induced by gamma radiation in the somatic cells of a radiosensitive mutant line of Drosophila melanogaster

Chromosome aberrations induced by gamma-rays in ganglia cells of Drosophila melanogaster larvae have been studied. Two strains of Drosophila were used: radiosensitive mutant rad (2) 201G1 and normal strain. It has been shown that the frequency of cells with chromosome aberrations in radiosensitive larvae is much more than in normal larvae after gamma-irradiation. The ratio of chromosome and chromatid deletions number to the number of exchange type aberrations is the same for both strains. The kinetics of chromosome aberrations induced in rad-larvae is similar to the normal one. The conclusion has been made that the realization of rad (2) 201G1 mutation takes place on the cell level.     

Genetics of the cell cycle: Adaptive modification of the <Emphasis Type="Italic">CycB</Emphasis><Superscript><Emphasis Type="Italic">2g</Emphasis></Superscript> mutation expression in dividing cells of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The effect of mutation CycB ^2g on mitosis in neural ganglia and imaginal disks was studied in third-instar larvae of Drosophila melanogaster. Chromosome condensation and segregation were shown to be impaired in dividing cells of mutant larvae. During the three-year period of maintenance of the mutation in heterozygote, frequencies of some defects decreased via cellular adaptive modification.Translated from Genetika, Vol. 41, No. 3, 2005, pp. 312–319.Original Russian Text Copyright © 2005 by Lebedeva, Trunova, Omelyanchuk.     

Regulator gene of phenyl oxidase activity in Drosophila melanogaster

A recessive lethal mutation l(2)hemo causing the occurrence of melanotic tumors in homozygous Drosophila larvae was found. The study of phenoloxidase (PO) activity revealed that the number of hemocytes with PO activity in homozygous larvae was significantly reduced (0.4 +/- 0.24%), compared to wild-type larvae (6.3 +/- 0.5%). On injury followed by injection with bacterial cells, the formation of melanotic thrombus did not occurred and hemocytes with PO activity were not recorded in homozygotes of line P103. Suppression of the activity of PO isozymes A1 and A3 was detected by means of electrophoretic analysis of homozygotes. According to gene mapping data, the localization of this mutation did not match any structural gene for known PO forms and is therefore related to a regulatory gene controlling the activity of the immune system of Drosophila.     

Experimental studies on a lethal gene (t) in the Mexican axolotl, Ambystoma mexicanum

In the Mexican axolotl, Ambystoma mexicanum, the developmental mutation lethal t is inherited as a simple Mendelian recessive. Mutant larvae failed to feed and died, on the average, 17 days after hatching. Unfed wild-type larvae died an average of 23 days after hatching. By 15 days, forelimb development had progressed further in the wild type; a cartilaginous scapula and humerus were present, but no cartilage was seen in the mutant limb. Histological examination indicated that the visceral cartilage may also be abnormal, and the rectus cervicus muscle was found to have fewer and smaller fibers. Though the mutant was not rescued by parabiosis with wild-type embryos, transplants of presumptive gill and limb tissue to wild-type hosts survived, indicating that the mutation is not an autonomous cell lethal.     

The behavior during the initial phase of in vitro aggregation of dissociated imaginal disc cells from Drosophila melanogaster

Aspects of the initial phase of aggregation in vitro of dissociated imaginal disc cells from Drosophila melanogaster are described. Using the methods described certain interdisc differences in the percent decrease in single cell units during the first hour of aggregation can be demonstrated. In addition it is shown that prospective notum cells isolated from the dorsal mesothoracic disc show less of a decrease than do prospective wing cells. This difference shows up in a variety of different wild-type stocks and in several mutant stocks as well. Prospective notum cells from the mutation fu59 show only a limited ability to adhere to one another, while the percentage of single cell units from prospective wing blade cells from r9 larvae grown on pyrimidine-poor medium decrease less compared to cells from the same stock grown on RNA-supplemented medium. There is a significantly greater decrease in the percent single cell units in cultures of prospective eye cells than in cultures of prospective antennal cells. Furthermore, cells from the antennal disc of larvae bearing the homeotic mutation ssa show a significantly lower decrease in single cell units when grown at restrictive temperatures. In contrast, antennal disc cells from the homeotic mutation ophthalmoptera; eyeless Dominant, a mutation which affects the eye disc, are unaffected, while cells from the eye disc are slightly less able to reassociate with one another.     

Age dynamics of the activity of the imago after chronic irradiation of larvae from the Drosophila line with disruption of apoptosis regulation

The relationship was studied between radiation-induced apoptosis in the nervous system of Drosophila larvae and the age dynamics in adult fly neuromuscular activity. The level of apoptosis in the neural ganglia of third-instar larvae from the wild-type strain increased 2.5 times after larval exposure to ionizing radiation (54 cGy). Irradiation of the strain with enhanced sensitivity to apoptosis induction, which carries a mutation in gene-inhibitor of apoptosis th (allele th4), and the wild-type strain Berlin led to an increase in neuromuscular activity of adult flies throughout the experiment and, consequently, to reduced aging rate. Conversely, this effect was not observed in strains with reduced sensitivity to induction of apoptosis (with mutations in genes dArk and Dcp-1).     

The effect of male mating competitiveness, developmental rate, and viability of larvae and pupae in Drosophila melanogaster heterozygous for the temperature-sensitive lethal mutation l(2)M167DTS on the dynamics of the mutation elimination from the population

The dependence of selection on an introduced mutation 1(2)M167DTS on male mating competitiveness, viability, and developmental rate of larvae and pupae of Drosophila melanogaster, heterozygous for this mutation, was examined in population experiments with preset conditions. The limitations of fitness parameters of individuals l(2)M167DTS/+ relative to individuals +/+ were estimated according to the conditions of the experiment and phenotypic characteristics of the mutation studied. Under conditions of limited food supply and dependence on emergence time in each generation, the sequence of female mating was shown to be of less importance than the order of medium utilization by the progeny of a certain genotype related to the male success in the first mating. The limiting factors acting on the l(2)M167DTS mutation were viability and developmental rate.     

Genotoxicity testing of five herbicides in the Drosophila wing spot test

Four triazine herbicides: amitrole, metribuzin, prometryn and terbutryn, and the bipyridal compound diquat dibromide have been evaluated for genotoxicity in the wing somatic mutation and recombination test of Drosophila melanogaster, following standard procedures. Third-instar larvae trans-heterozygous for the third chromosome recessive markers multiple wing hairs (mwh) and flare-3 (flr(3)) were chronically fed with different concentrations of the test compounds. Feeding ended with pupation of the surviving larvae. Genetic changes induced in somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage and mitotic recombination produce single spots; while twin spots are produced only by mitotic recombination. Exposure to 0.5 mM and 1 mM of amitrole clearly increased the frequency of small single, large single and total spots. Terbutryn, at the concentration of 5 mM, induced a slight increase in the frequency of small single and total spots, but this result could be false positive. The other three herbicides tested did not show any genotoxic effect. When heterozygous larvae for mwh and the multiple inverted TM3 balancer chromosomes were treated, significant increases in the frequency of mutant spots were only detected for amitrole. The observed spot frequencies were lower than those found in mwh/flr(3)50%) of the total spot induction was due to mitotic recombination.     

Zebrafish fat-free is required for intestinal lipid absorption and Golgi apparatus structure

The zebrafish fat-free (ffr) mutation was identified in a physiological screen for genes that regulate lipid metabolism. ffr mutant larvae are morphologically indistinguishable from wild-type sibling larvae, but their absorption of fluorescent lipids is severely impaired. Through positional cloning, we have identified a causative mutation in a highly conserved and ubiquitously expressed gene within the ffr locus. The Ffr protein contains a Dor-1 like domain typical of oligomeric Golgi complex (COG) gene, cog8. Golgi complex ultrastructure is disrupted in the ffr digestive tract. Consistent with a possible role in COG-mediated Golgi function, wild-type Ffr-GFP and COG8-mRFP fusion proteins partially colocalize in zebrafish blastomeres. Enterocyte retention of an endosomal lipid marker in ffr larvae support the idea that altered vesicle trafficking contributes to the ffr mutant defect. These data indicate that ffr is required for both Golgi structure and vesicular trafficking, and ultimately lipid transport.     

The vesicular integral protein-like gene is essential for development of a mechanosensory system in zebrafish

The zebrafish hi472 mutation is caused by a retroviral insertion into the vesicular integral protein-like gene, or zVIPL, a poorly studied lectin implicated in endoplasmic reticulum (ER)-Golgi trafficking. A mutation in the shorter isoform of zVIPL (zVIPL-s) results in a reduction of mechanosensitivity and consequent loss of escape behavior. Here we show that motoneurons and hindbrain reticulospinal neurons, which normally integrate mechanosensory inputs, failed to fire in response to tactile stimuli in hi472 larvae, suggesting a perturbation in sensory function. The hi472 mutant larvae in fact suffered from a severe loss of functional neuromasts of the lateral line mechanosensory system, a reduction of zVIPL labeling in support cells, and a reduction or even a complete loss of hair cells in neuromasts. The Delta-Notch signaling pathway is implicated in cellular differentiation of neuromasts, and we observed an increase in Notch expression in neuromasts of hi472 mutant larvae. Treatment of hi472 mutant larvae with DAPT, an inhibitor of Notch signaling, or overexpression of the Notch ligand deltaB in hi472 mutant blastocysts produced partial rescue of the morphological defects and of the startle response behavior. We conclude that zVIPL-s is a necessary component of Delta-Notch signaling during neuromast development in the lateral line mechanosensory system.     

Developmentally regulated alternate modes of expression of the Gpdh locus of Drosophila.

Immunoblot analyses have been performed on extracts prepared from Drosophila melanogaster. Those analyses have revealed two subunit forms of enzyme glycerol 3-phosphate dehydrogenase (GPDH) in larval tissues and in adult abdominal tissues. Thoracic tissue, which accounts for the bulk of the adult GPDH, has only one subunit form, the smaller. The two subunit forms differ by approximately 2400 daltons. In agreement with previous genetic and biochemical data indicating that this enzyme is encoded by a single structural gene, analyses of extracts prepared from a strain carrying a GPDH null mutation detect no GPDH polypeptides in larvae or adults. Similarly, analyses of extracts prepared from a strain carrying a mutation which produces a GPDH polypeptide that differs in size from wild-type reveal a change in the adult thoracic GPDH polypeptide as well as a change in both GPDH polypeptides found in larvae. Total Drosophila RNA prepared from larvae or newly eclosed adults has been translated in a mRNA-dependent cell-free system. GDPH was immunoprecipitated from the translation products and analyzed. Two subunit forms of GPDH were immunoprecipitated from translation products whose synthesis was directed by larval RNA and only one was detected in the polypeptides synthesized from adult RNA. The GPDH polypeptides synthesized in vitro are approximately the same size as the corresponding polypeptides found in vivo. The relative proportion of total GPDH represented by each subunit form synthesized in vitro is similar to those found in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)     

Involvement of eddy currents in the mutagenicity of ELF magnetic fields

Possible carcinogenic and/or mutagenic activity of extremely low frequency magnetic fields was examined using somatic mutation and recombination test system of Drosophila melanogaster. An X-linked semi-dominant DNA repair defective mutation mei-41(D5) was introduced into the conventional mwh/flr test system to enhance mutant spot frequency. Virgin females of w mei-41(D5)/FM6; flr/TM6 were crossed with w mei-41(D5)/Y; mwh jv; spa(pol) males. The F(1) third instar larvae were exposed to a 50Hz, 20mT sinusoidal AC magnetic field for 24h. After moulting from pupal cases, their wings were examined under a bright field microscope to detect hair spots with mwh or flr mutant morphology. The exposure caused a statistically significant enhancement in somatic recombination spot frequency. Mutant spots arising due to chromosomal non-disjunction or terminal deletion also increased but the frequency of spots resulting from point mutation was not altered. The enhancement in the recombination spot frequency was suppressed to the control level when a culture medium without electrolytes was used during exposure. When larvae were exposed to a magnetic field in an annular dish, flies from the outer ring showed more mutant spots compared to those from the inner ring. These results suggest that the detected mutagenic activity was that of the induced eddy current, rather than that of the magnetic field itself.     

Ivory: a recessive white-eyed tryptophan metabolism mutant with intermediate F2 — and R1 — progenies in the flesh fly Sarcophaga barbata

Summary The recessive autosomal gene ivory (i) causes white eyes in the flesh fly Sarcophaga barbata. The mutation completely blocks the synthesis of formylkynurenine. Ivory larvae and ivory imagoes are able to synthesize xanthommatin if formylkynurenine and kynurenine respectively are provided with the food. The eye colour of the F₂- and the R₁-mutants respectively is intermediate because these animals have taken up xanthommatin precursors that were excreted by the wild-type larvae. The white eye colour is not influenced by temperature. The viability of the ivory stock is somewhat lower than that of the wild-type stock. The mutation ivory is homologous to the mutation vermilion of Drosophila melanogaster and to the mutation green of Musca domestica respectively.     

Genotoxicity of neutrons in Drosophila melanogaster. Somatic mutation and recombination induced by reactor neutrons

This paper describes the observation of a direct relationship between the absorbed doses of neutrons and the frequencies of somatic mutation and recombination using the wing somatic mutation and recombination test (SMART) of Drosophila melanogaster. This test was used for evaluating the biological effects induced by neutrons from the Triga Mark III reactor of Mexico. Two different reactor power levels were used, 300 and 1000 kW, and two absorbed doses were tested for each power level: 1.6 and 3.2 Gy for 300 kW and 0.84 and 1.7 Gy for 1000 kW. A linear relationship was observed between the absorbed dose and the somatic mutation and recombination frequencies. Furthermore, these frequencies were dependent on larval age: In 96-h-old larvae, the frequencies were increased considerably but the sizes of the spots were smaller than in 72-h-old larvae. The analysis of the balancer-heterozygous progeny showed a linear absorbed dose- response relationship, although the responses were clearly lower than found in the marker-trans-heterozygous flies. Approximately 65% of the genotoxicity observed is due to recombinational events. The results of the study indicate that thermal and fast neutrons are both mutagenic and recombinagenic in the D. melanogaster wing SMART, and that the frequencies are dependent on neutron dose, reactor power, and the age of the treated larvae.