Mutation of Indy(p115) increases the life expectancy of imago of Drosophila melanogaster depending on sex and genetic background

The Indyp115 allele in heterozygous state almost doubles the life span of adult Drosophila melanogaster, and this effect largely depends on the strain used for obtaining heterozygotes. Male and female life span depends on Indyp115 to a different degree. Apart from Indyp115 heterozygotes, sexual dimorphism for life span was also observed in strain Hikone-AW, but not in OregonR and TM3 balancer. In heterozygotes Indyp115/OR, both the average life span and female reproductive period increased as compared to OR control. No substantial increase in female reproductive period in Indyp115/Hk heterozygotes was found. In homozygotes for allele Indyp115, we have previously revealed two phases of embryotic lethality and lethality at the larval and pupa stages. Thus, allele Indyp115 has a double and opposite effect on Drosophila viability. On the one hand, it extends the life span of adult flies, on the other, decreases survival at preimaginal stages.     

Identifying the Indyp115 Mutation of the Na+-Dicarboxylate Transporter Gene in Drosophila melanogaster

The P[lArB]-element insertional mutation (Indy ^p115) was obtained in the promoter region of the Na⁺-dicarboxylate transporter gene of Drosophila (geneIndy, I'm not dead yet) within the 75D region of chromosome 3. The expression pattern of the reporter -galactosidase was determined in various tissues of third-instar larvae and adult flies. Both males and females homozygous for this mutation were fertile, though their viability was reduced. The two lethality phases were revealed in embryogenesis at nucleus cleavage stages 1–5 to the point of blastoderm formation and at latest stages 15–16, as well as at stage 17, when the larval development is completed though the larva still remains in the chorion. The gene expression in the follicular cells of an embryo at the terminal oogenesis stages is suggested to cause the first lethality stage. The expression pattern of this gene seems also to account for the tissue- and stage-specific activity of the 5"-regulatory region in the Indy gene.     

Effects on kidney disease, fertility and development in mice inheriting a protein-truncating Denys-Drash syndrome allele (Wt1 tmT396)

Denys-Drash syndrome (DDS) is caused by heterozygous mutations of the Wilms’ tumour suppressor gene, WT1, characterised by early-onset diffuse mesangial sclerosis often associated with male pseudohermaphroditism and/or Wilms’ tumourigenesis. Previously, we reported that the Wt1 ^tmT396 allele induces DDS kidney disease in mice. In the present study heterozygotes (Wt1 ^tmT396/+) were generated on inbred (129/Ola), crossbred (B6/129) and MF1 second backcross (MF1-N2) backgrounds. Whereas male heterozygotes on each background were fertile, inbred heterozygous females were infertile. Kidney disease (proteinuria and sclerosis) was not congenital and developed significantly earlier in inbred mice, although with variable onset. Disease onset in MF1-N2 stocks occurred later in Wt1 ^tmT396/+ mice than reported previously for Wt1 ^R394W/+ mice, and while no kidney disease has been reported in B6/129 Wt1 ^+/- mice, B6/129 Wt1 ^tmT396/+ mice were affected. Offspring of both male and female B6/129 and MF1-N2 Wt1 ^tmT396/+ mice developed kidney disease, but its incidence was significantly higher in offspring of female heterozygotes. Wt1 ^{tmT396/tmT396} embryos exhibited identical developmental abnormalities to those reported for Wt1 ^-/- embryos. The results indicate that the Wt1 ^tmT396 allele does not predispose to Wilms’ tumourigenesis or male pseudohermaphroditism, its effect on kidney disease and female fertility depends on genetic background, stochastic factors may affect disease onset, and disease transmission is subject to a partial parent-of-origin effect. Since the Wt1 ^tmT396 allele has no detectable intrinsic functional activity in vivo, and kidney disease progression is affected by the type of Wt1 mutation, the data support the view that DDS nephropathy results from a dominant-negative action rather than WT1 haploinsufficiency or gain-of-function.     

Sterilizing Effect of the Mutant Allele sbr 10 (l(1)ts403) in Compound with Null Allele in Drosophila melanogaster Females

In females of Df(1)v-L4/+(0/+) genotype, the presence of the wild-type allele ofsmall bristles (sbr) gene in a single dose has no significant effect on their fecundity, whereas a reduced dose of the temperature-sensitive allele sbr ¹⁰ (l(1)ts403) causes a strong sterilizing effect in females Df(1)v-L4/sbr ¹⁰ (0/sbr ¹⁰) at permissive temperature. We studied the contribution to this effects of the following factors: resorption of egg chambers, decreased oviposition, offspring death at the embryonic and larval stages, and reduced fecundity in females 0/sbr ¹⁰. Sterilizing effect of the mutant sbr ¹⁰ allele proved to be primarily caused by offspring lethality at the embryonic and first-instar larval stages. In 0/+ females, the majority of undeveloped eggs contained embryos that perished at the late developmental stages, whereas in females 0/sbr ¹⁰, at least 50% of undeveloped egg showed no visible signs of development or the embryo development was arrested at early stages of embryogenesis. The results obtained suggest insufficiency of the temperature-sensitive allele sbr ¹⁰ in haploid state to ensure the reproductive functions of Drosophila melanogaster females.     

Murine “housekeeping” enzyme (genetic locus:Idh-1) is regulated in an allele-specific manner

The murine “housekeeping” enzyme, cytosolic NADP-isocitrate dehydrogenase (E.C. 1.1.1.42) (genetic locus:Idh-1), exhibited a complex pattern of allele-specific expression. Protein electrophoresis on cellulose-acetate gels and determination of relative enzymatic activity by means of densitometry revealed that in heart tissue (but not liver tissue) of certain hybrid crosses the AA-homodimer was underrepresented relative to total enzymatic activity, and the degree of underrepresentation changed during development. In mixtures of homozygous tissue extracts of heart tissue (but not liver tissue) the AA-homodimer was underrepresented relative to the BB-homodimer. Relative activity of allelic isozymes varied as a function of tissue (heart versus liver), age, and the parental source of the Idh-1^a allele, but did not vary as a function of sex. Allele-specific expression was also exhibited in kidney tissue of the same animals. In adult male kidney tissue extracts from heterozygotes, the AA-hornodimer was underrepresented relative to total enzymatic activity; in adult female kidney tissue extracts from heterozygotes, a more codominant phenotype was observed. Tissue extracts from immature hybrid animals exhibited a phenotype midway between the adult male and adult female phenotypes. Tissue extracts from castrated males exhibited a phenotype equivalent to that seen in females. Relative activity of allelic isozymes in kidney varied as a function of age and sex, but did not vary as a function of the parental source of the Idh-1^a allele. While cytosolic NADP-IDH is a “housekeeping” enzyme, expressed in multiple tissues of the mouse, differences in the relative intensities of allelic isozyme bands provide evidence for tissue- and stage-specific regulatory variation.     

Reproductive features of the non-native Siganus luridus (Teleostei, Siganidae) during early colonization at Linosa Island (Sicily Strait, Mediterranean Sea)

In July 2003, the finding of a newly settled population of Siganus luridus at Linosa Island (Sicily Strait, Mediterranean Sea) gave us the unusual opportunity to examine the reproductive condition of a Lessepsian migrant during early phases of colonization. Aspects of gonad morphology, fecundity, atresia and oocyte dynamics were investigated by using 43 pioneer specimens collected in concomitance with their first record in the Pelagie Islands. Ovarian development was consistent with the group‐synchronous type, and testicular organization was of the unrestricted spermatogonial testis type, with cystic spermatogenesis. Both males and females had reached final stages of gonad maturation. The rates of follicular atresia were moderate: out of 17 adult females, 10 individuals did not present atretic oocytes; six exhibited <15.1% of secondary growth phase (SGP) oocytes in α‐atresia, while one female presented 45.7% of SGP in α‐atresia. Fecundity estimates did not diverge from what was observed in a reference population along the Lebanese coast. Absolute fecundity ranged from 115 739 to 740 433 oocytes per female (16.5–24.5 cm L_T). Relative fecundity ranged from 1239 to 3162 oocytes g⁻¹, with a mean of 1885 ± 868 oocytes g⁻¹. Our observations indicated that these early settled siganids are reproductively active at Linosa and suggested the forthcoming of self‐maintaining populations across the central Mediterranean area.     

Impaired RIPK1 ubiquitination sensitizes mice to TNF toxicity and inflammatory cell death

Receptor-interacting protein 1 (RIP1; RIPK1) is a key regulator of multiple signaling pathways that mediate inflammatory responses and cell death. TNF-TNFR1 triggered signaling complex formation, subsequent NF-κB and MAPK activation and induction of cell death involve RIPK1 ubiquitination at several lysine residues including Lys376 and Lys115. Here we show that mutating the ubiquitination site K376 of RIPK1 (K376R) in mice activates cell death resulting in embryonic lethality. In contrast to Ripk1^K376R/K376R mice, Ripk1^K115R/K115R mice reached adulthood and showed slightly higher responsiveness to TNF-induced death. Cell death observed in Ripk1^K376R/K376R embryos relied on RIPK1 kinase activity as administration of RIPK1 inhibitor GNE684 to pregnant heterozygous mice effectively blocked cell death and prolonged survival. Embryonic lethality of Ripk1^K376R/K376R mice was prevented by the loss of TNFR1, or by simultaneous deletion of caspase-8 and RIPK3. Interestingly, elimination of the wild-type allele from adult Ripk1^K376R/cko mice was tolerated. However, adult Ripk1^K376R/cko mice were exquisitely sensitive to TNF-induced hypothermia and associated lethality. Absence of the K376 ubiquitination site diminished K11-linked, K63-linked, and linear ubiquitination of RIPK1, and promoted the assembly of death-inducing cellular complexes, suggesting that multiple ubiquitin linkages contribute to the stability of the RIPK1 signaling complex that stimulates NF-κB and MAPK activation. In contrast, mutating K115 did not affect RIPK1 ubiquitination or TNF stimulated NF-κB and MAPK signaling. Overall, our data indicate that selective impairment of RIPK1 ubiquitination can lower the threshold for RIPK1 activation by TNF resulting in cell death and embryonic lethality.Subject terms: Acute inflammation, Chronic inflammation     

Genetic and developmental variation of hemoglobin in the deermouse,Peromyscus maniculatus

A genetic investigation of electrophoretic hemoglobin variants of the deermouse, Peromyscus maniculatus, shows three alleles, HbI ^f, HbI^r, and HbI ^o, at a duplicated site controlling the six adult phenotypes. The HbI ^fallele has not been described previously. The hemoglobin locus is not closely linked to the albino locus. Fetal hemoglobin is distinct from any of the adult components and has a slower electrophoretic mobility. The fetal phenotype changes to the adult type between the days 15 and 18 of prenatal life.     

Interactions between enhancer of rudimentary and Notch and deltex reveal a regulatory function of enhancer of rudimentary in the Notch signaling pathway in Drosophila melanogaster

Enhancer of rudimentary, e(r), encodes a small nuclear protein, ER, that has been implicated in the regulation of pyrimidine metabolism, DNA replication, and cell proliferation. In Drosophila melanogaster, a new recessive Notch allele, N^nd-p, was isolated as a lethal in combination with an e(r) allele, e(r)^p2. Both mutants are viable as single mutants. N^nd-p is caused by a P-element insertion in the 5' UTR, 378-bp upstream of the start of translation. Together the molecular and genetic data argue that N^nd-p is a hypomorphic allele of N. The three viable notchoid alleles, N^nd-p, N^nd-1, and N^nd-3, are lethal in combination with e(r)^- alleles. Our present hypothesis is that e(r) is a positive regulator of the Notch signaling pathway and that the lethality of the Ne(r) double mutants is caused by a reduction in the expression of the pathway. This is supported by the rescue of the lethality by a mutation in Hairless, a negative regulator of N, and by the synthetic lethality of dxe(r) double mutants. Further support for the hypothesis is a reduction in E(spl) expression in an e(r)^- mutant. Immunostaining localizes ER to the nucleus, suggesting a nuclear function for ER. A role in the Notch signaling pathway, suggests that e(r) may be expressed in the nervous system. This turns out to be the case, as immunostaining of ER shows that ER is localized to the developing CNS.     

Moms are better nurses than dads: gender biased self‐facilitation in a dioecious Juniperus tree

Questions: Can gender of nurse plants affect regeneration patterns and spatial population structure? Is there a seed‐seedling conflict in the regeneration process? What factors are responsible for the clumped spatial population structure observed for adult trees? Location: Mediterranean cold semi‐arid high mountains in Spain. Methods: The spatial pattern of adult Juniperus thurifera trees was studied by means of Ripley's K‐analysis. χ² analyses were used to test for natural seedling frequency in each of three main microhabitats: (1) under female and (2) male tree canopies and (3) in open interspaces. The observed pattern was explained experimentally by studying seed and seedling survival for two years. Survival probabilities were calculated across life stages for each of three main microhabitats. Results: Adult J. thurifera trees were aggregated in space. Most seedlings were found underneath female J. thurifera trees. Experimental studies demonstrated that from seed dispersal to seedling survival all life stages showed the same positive or negative trend within a given microhabitat, indicating stage coupling and no seed‐seedling conflicts. Attraction of frugivo‐rous birds by reproductive female junipers and improvement of environmental conditions beneath tree canopies were the main factors responsible for the variation in seedling density among microhabitats; highest underneath female trees and lowest in open interspaces. Conclusions: In dioecious species, the gender of nurse plants can significantly determine the spatial population structure. In J. thurifera forests, facilitation beneath female trees occurs among all life stages without any sign of seed‐seedling conflict. The most critical factors shaping the spatial population structure were directed seed dispersal and environmental amelioration beneath female conspecific trees.     

Association between 242C>T polymorphism of NADPH oxidase p22phox gene (CYBA) and longevity in Russian population

Life span depends on many factors, including the level of reactive oxygen species, like superoxide radical. Superoxide radical is produced from oxygen in the course of the oxidation of NADPH to NADP⁺. The process is catalyzed by NADPH oxidase. In this study, genotype and allele distributions of the C242T (rs4673) polymorphism in the CYBA gene, which encodes the α subunit of NADPH oxidase (p22phox), were examined in the sample of long livers and in the population sample of the city of Tomsk. Statistically significantly higher frequency of T allele among female long livers (34.625%), compared to the females from Russian population (26.32%) was demonstrated (χ² = 5.226; p = 0.022; OR = 1.48). Thus, the T allele is associated with a high life expectancy in females from the Russian population. No such association was observed for males from the same population.     

No influence of Indy on lifespan in Drosophila after correction for genetic and cytoplasmic background effects

To investigate whether alterations in mitochondrial metabolism affect longevity in Drosophila melanogaster, we studied lifespan in various single gene mutants, using inbred and outbred genetic backgrounds. As positive controls we included the two most intensively studied mutants of Indy, which encodes a Drosophila Krebs cycle intermediate transporter. It has been reported that flies heterozygous for these Indy mutations, which lie outside the coding region, show almost a doubling of lifespan. We report that only one of the two mutants lowers mRNA levels, implying that the lifespan extension observed is not attributable to the Indy mutations themselves. Moreover, neither Indy mutation extended lifespan in female flies in any genetic background tested. In the original genetic background, only the Indy mutation associated with altered RNA expression extended lifespan in male flies. However, this effect was abolished by backcrossing into standard outbred genetic backgrounds, and was associated with an unidentified locus on the X chromosome. The original Indy line with long-lived males is infected by the cytoplasmic symbiont Wolbachia, and the longevity of Indy males disappeared after tetracycline clearance of this endosymbiont. These findings underscore the critical importance of standardisation of genetic background and of cytoplasm in genetic studies of lifespan, and show that the lifespan extension previously claimed for Indy mutants was entirely attributable to confounding variation from these two sources. In addition, we saw no effects on lifespan of expression knockdown of the Indy orthologues nac-2 and nac-3 in the nematode Caenorhabditis elegans.     

Influence of <Emphasis Type="Italic">Lecanicillium attenuatum</Emphasis> on the development and reproduction of the cotton aphid, <Emphasis Type="Italic">Aphis gossypii</Emphasis>

The activity of entomopathogens on insect pests has been investigated for many species but the influence of entomopathogenic fungi on factors other than mortality relating to population increase has not been frequently studied. The influence of Lecanicillium attenuatum CS625 (=Verticillium lecanii CS625) on development and reproduction of cotton aphid (Aphis gossypii) was investigated. A conidia suspension of the isolate was applied onto first instar nymphs. Increased spore concentration did not significantly affect each nymphal stage, total nymphal period, pre-reproductive period and the age of first larviposition. A significant dose effect on reduction of life span, reproductive period and fecundity was observed in 1st and 3rd instars after spore application. When conidia were applied to 1st instars, life span was significantly reduced to 10.8 and 8.4 days at 1 × 10⁴ and 1 × 10⁸ conidia/ml, respectively from 12.2 days in the control. During the life span, total fecundity was 41 ± 7.3, 26 ± 0.8 and 22 ± 5.7 nymphs per female at 1 × 10⁴, 1 × 10⁶ and 1 × 10⁸ conidia/ml, respectively compared with 51 ± 2.0 nymphs per untreated female. Reproduction period was also significantly shortened with increasing spore concentration. Application of spores to 3rd instars showed a similar trend. However, daily fecundity of individual aphids was not affected by spore dose. It was concluded that the isolate of L. attenuatum is able to affect populations of cotton aphid by reducing life span and total fecundity as well as by killing the aphids directly.     

The Effect of Food Concentration on the Life History of Three Types of Brachionus calyciflorus Females

The effect of food concentration on the life history of three types of Brachionus calyciflorus females (amictic, unfertilized mictic and fertilized mictic female) was studied with replicated individual cultures at 25 ° and at four food concentrations (1.5, 3.0, 6.0 and 9.0 × 10⁶ cells mL^—1) of Scenedesmus obliquus. There were highly significant effects of both food concentration and female type, independently and in interaction on the duration of juvenile period of the rotifer, but neither were the effects on the duration of post‐reproductive period and mean life‐span. The duration of juvenile period of unfertilized mictic female at the food concentration of 9.0 × 10⁶ cells mL^—1 was the longest among all the food concentration‐female type combinations. Both food concentration and female type influenced significantly the duration of reproductive period and the number of eggs produced by each type of female per life cycle, respectively. There was, however, no significant interaction between food level and female type. Among the three types of females, the number of eggs produced by an unfertilized mictic female was the largest, and that of a fertilized mictic female was the smallest.     

Cancer and aging: More puzzles,more promises?

In this issue of Cell Cycle, a new paper shows that metformin, an oral antidiabetic drug that activates AMP-activated protein kinase, prolongs both mean and maximal life span and prevents reproductive aging of female mice. Unexpectedly, metformin did not decrease the incidence of cancer in this mice strain. Here, we discuss the relationship between aging and cancer, the mechanism of metformin action, and the prospects of using this compound for life span extension in humans.     

Temperature-dependent development and reproductive traits of <Emphasis Type="Italic">Tetranychus macfarlanei</Emphasis> (Acari: Tetranychidae)

Development and reproductive traits of Tetranychus macfarlanei Baker & Pritchard (Acari: Tetranychidae) were investigated on kidney bean, Phaseolus vulgaris L., at eleven constant temperatures. Tetranychus macfarlanei was able to develop and complete its life cycle at temperatures ranging from 17.5 to 37.5°C. At 15 and 40°C, a few eggs (2–4%) hatched but further development was arrested. Development from egg to adult was slowest at 17.5°C and fastest at 35°C for both females and males. Using Ikemoto and Takai’s linear model, the estimated lower developmental thresholds for egg-to-female adult, egg-to-male adult and egg-to-egg development were 12.9–13.0°C. The thermal constants for the respective stages were 110.85, 115.99 and 125.32 degree-days (DD). The intrinsic optimum temperatures (T _Φ) calculated by non-linear SSI model were determined as 24.4, 24.4 and 24.2°C for egg-to-female adult, egg-to-male adult and egg-to-egg development, respectively. The net reproductive rate (R ₀) was highest at 25°C (167.4 females per female) and lowest at 17.5°C (42.6 females per female). The intrinsic rate of natural increase, r _m, increased linearly with the rising of temperature from 0.102 at 17.5°C to 0.441 day⁻¹ at 35°C. These values suggested that T. macfarlanei could be growing quickly in response to increasing temperatures from 17.5 to 35°C and provide a basis for predicting its potential geographical range.     

Demographic traits of Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae) reared on Aphis fabae Scopoli (Hemiptera: Aphididae)

Lady beetles are the most important predatory species among arthropods; thus, studying their population parameters gives a clear picture about their life span. The demography of the lady beetle, Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae), was studied under laboratory condition, on Aphis fabae at 27?°C based on the age-stage, two-sex life table. The means and standard errors were estimated by the bootstrap techniques. The results showed that the intrinsic rate of increase (r) and the finite rate of increase (λ) were 0.18 and 1.20 d^?1, respectively. The net reproduction rate (R ₀) and the gross reproduction rate were 232.49 and 442.48 offspring/individual, respectively. The mean generation time (T) was 29.03?days. The first peak of reproductive value in females was on the 18th day, which coincides with the total pre-oviposition period counted from birth. The life expectancy curve (e_xj ) of a new hatched female adult was 60?days. The results demonstrated that growth, survival and development of the predator can be described accurately by use of the age-stage, two-sex life table.     

Methuselah meets diabetes

Mutations in the daf-2 and age-1 genes cause constitutive dauer larva formation and double adult life span in C. elegans. Their effect on life span has excited considerable interest and their effect on dauer formation has facilitated rapid progress in their genetic and molecular analysis. Two recent papers^12,13 report that daf-2 encodes a member of the insulin-receptor family and that age-1 encodes a PI3 kinase subunit, a second-messenger producing enzyme known to act downstream of the mammalian insulin receptor. These findings provide the first mechanistic insight into the well-established link between metabolism and aging. BioEssays 20:113–115, 1998. © 1998 John Wiley & Sons, Inc.     

The effect of adult feeding on the fecundity,weight loss and survival of the pine beauty moth,Panolis flammea (D&S)

Summary Adult females of the pine beauty moth, Panolis flammea, fed on a saturated honey solution, laid significantly more eggs than those individuals given water only or not fed at all.At 20°, the longevity of both male and female moths was significantly increased when they were supplied with a food source. At 10° fed male months were not significantly longer lived than unfed moths. Fed female moths at 10° were however, significantly longer lived than unfed individuals. At 20°, fed male moths lived longer than fed females (8.17 days as compared with 5.95 days). At 10°, fed female moths lived longer than fed males (16.00 days compared with 11.41 days).At 20°, weight loss in unfed females over the reproductive period was proportional to the number of eggs laid and the survival time of the individual. In fed females, no significant weight loss occurred over the reproductive period, irrespective of the number of eggs laid or the longevity of the individual.Egg production, even in fed females, over the life span of an individual was only a fifth of the total eggs present in the reproductive tract at eclosion.It is concluded that adult females of P. flammea are likely to require an extra external food source to achieve their full reproductive potential.