Heterologous interchange and nondisjunction of distributively paired chromosomes in Drosophila melanogaster mature oocytes

In the mature oocyte C(1)-4 and Y-4 interchanges do not direct disjunction of the centromeres of the involved heterologs. C(1)-4 detachments are recovered in either sex and almost every time ($\text{13}\text{14}$) with a free fourth chromosome that is the sister of the involved four. Y-4 fragments are recovered preferentially in males ($\text{28}\text{28}$) and with a free, sister four ($\text{26}\text{28}$). In the mature oocyte segregation patterns are determined and C(1)-4 and Y-4 interchange usually involve heterologs segragating to the same pole.     

A rapid selection technique for detecting meiotic X-chromosomal nondisjunction in Drosophila melanogaster

A chemical selection technique is described for the rapid and easy detection of X-chromosomal nondisjunction in females of Drosophila melanogaster. The method employs the maroon-like (ma-1) gene and depends on the known hypersensitivity of ma-1 flies lacking xanthine dehydrogenase (XDH) activity to killing by treatments with aqueous purine solutions. Parental females, heterozygous for two ma-1 alleles which produce 25% of wild-type XDH activity, are mated to males bearing a non-complementing ma-1 allele. After treatment of developing cultures with a 6-mM purine solution, only those individuals possessing 25% or greater XDH activity survive to eclosion. The present report demonstrates that this system can be used to measure accurately the spontaneous frequency of X-chromosomal nondisjunction as well as increased maternal nondisjunction produced by cold treatment, X-irradiation or meiotic mutants. The rapidity and ease of this system suggest that it can be used for the routine monitoring of environmental agents for those which produce this class of meiotic segregational anomalies.     

Variations in the amount of polysomes in mature oocytes of Drosophila melanogaster.

Quantitative measurements of polysomes and ribosomes of Drosophila melanogaster egg chambers, mature oocytes, and embryos were done using sucrose gradient analysis. The amount of polysomes per egg chamber increases about 20 times from stage 5 to 13, and then remains constant up to the end of embryogenesis. The percentage of ribosomes in polysomes is fairly constant during oogenesis and embryogenesis (56 ± 7%). Depending on the fly population, the percentage of ribosomes in polysomes of mature oocytes varies from 10 to 70%. It is shown that the percentage of polysomes in mature oocytes decreases with the time of retention of the mature oocytes in the ovary. Twenty-four- to thirty-six-hour-old flies kept in optimal conditions retain their mature oocytes for 2–3 hr. These mature oocytes still contain 40–60% ribosomes in polysomes. Conditions are given which allow the obtainment of reproducibly high amounts of polysomes from mature oocytes of Drosophila.     

Cytology of aberrations induced by X-rays in oocytes of Drosophila melanogaster.

200 first-division configurations were analyzed for cytological aberrations induced by X-rays in late meiotic prophase in oocytes of Drosophila melanogaster. For the 3000 and 6000 r doses, 38 and 66%, respectively, were classified as abnormal. The aberrant divisions included displacement of the chromosomes suggesting their non-disjunction, loss of a whole chromosome, fragments and heterologous exchanges and unidentifiable aberrations. Non-disjunctional chromosomes were free of heterologous exchanges. The concept that a majority of X-ray-induced dominant lethals is due to chromosomal breakage is supported by the findings of the present study.     

The mechanism of secondary nondisjunction in Drosophila melanogaster females

Bridges (1916) observed that X chromosome nondisjunction was much more frequent in XXY females than it was in genetically normal XX females. In addition, virtually all cases of X nondisjunction in XXY females were due to XX <--> Y segregational events in oocytes in which the two X chromosomes had failed to undergo crossing over. He referred to these XX <--> Y segregation events as "secondary nondisjunction." Cooper (1948) proposed that secondary nondisjunction results from the formation of an X-Y-X trivalent, such that the Y chromosome directs the segregation of two achiasmate X chromosomes to opposite poles on the first meiotic spindle. Using in situ hybridization to X and YL chromosomal satellite sequences, we demonstrate that XX <--> Y segregations are indeed presaged by physical associations of the X and Y chromosomal heterochromatin. The physical colocalization of the three sex chromosomes is observed in virtually all oocytes in early prophase and maintained at high frequency until midprophase in all genotypes examined. Although these XXY associations are usually dissolved by late prophase in oocytes that undergo X chromosomal crossing over, they are maintained throughout prophase in oocytes with nonexchange X chromosomes. The persistence of such XXY associations in the absence of exchange presumably facilitates the segregation of the two X chromosomes and the Y chromosome to opposite poles on the developing meiotic spindle. Moreover, the observation that XXY pairings are dissolved at the end of pachytene in oocytes that do undergo X chromosomal crossing over demonstrates that exchanges can alter heterochromatic (and thus presumably centromeric) associations during meiotic prophase.     

Embryonic and post-embryonic lethals induced by diethyl sulfate in mature sperm of Drosophila melanogaster.

It has recently been reported that, in Drosophila melanogaster, when sperm treated with diethyl sulfate was stored in the females, II–III translocations were detected as from the 6th day after the treatment, though none was recovered without storage. Chromosome breaks being currently considered the main cause of dominant lethality and the embryonic period lasting about one day at 25°C, it was thought of interest to study the ability of DES to induce this type of damage with and without storage. It was found that the treatment increased embryonic lethality (measured as frequency of unhatched eggs) and post-embryonic lethality (measured as frequency of larval and pupal death) over the control values. The frequency of embryonic lethals after storage in the females for 6 days was similar to that shown by the unstored samples. In contrast with this, the yield of post-embryonic lethality was markedly raised by that storage time. It is suggested that: (1) lesions are induced as “pre-breaks”, and storage and cell divisions are instrumental in their opening; (2) potential breaks can undergo DNA replication and cell division as such and become open in different cell cycles, impairing embryonic and post-embryonic development; (3) chromosome breaks induced by DES seem to behave in a way similar to those induced by other mono- and poly-functional alkylating agents; and (4) when the potential ability of chemical compounds to induce chromosome breaks is assessed, post-embryonic lethality can be used as a simple one-generation preliminary test, to establish delayed effects.     

In vitro induced pinocytotic activity by a juvenile hormone analogue in oocytes of Drosophila melanogaster

Summary Pinocytotic activity has been analyzed in Drosophila oocytes following either in vivo or in vitro exposure to horseradish peroxidase. The enzyme tracer gains access to the yolk spheres only when supplied to the oocyte in vivo. In oocytes cultured in vitro, peroxidase remains restricted to the residual coated vesicles and to the tubular profiles formed in excess in the cortical ooplasm.In an attempt to induce peroxidase uptake by oocytes cultured in vitro, various incubations were tested. Among these, hemolymph from both sexes is capable of promoting peroxidase uptake up to a level comparable to that detectable in vivo. On the other hand, fat body extracts fail to promote such cellular activity. Finally, the juvenile hormone analogue ZR-515 is shown to be the only factor required to promote pinocytotic activity under the experimental conditions tested. The observations are interpreted to indicate that vitellogenin has no inductive role on pinocytosis but simply acts by adhering to the forming coated vesicles which in turn are produced by the oolemma in response to the action of juvenile hormone.     

Ecdysteroids regulate yolk protein uptake by Drosophila melanogaster oocytes

Juvenile hormones (JHs) are thought to drive the regulation of yolk protein uptake by ovaries in Drosophila melanogaster. However, the level of JH production in a mutant stock (ap(56f)) is depressed yet the flies are normally vitellogenic. The production of ecdysteroids by these ap(56f) ovaries in vitro is elevated above that of wild-type ovaries. The incubation of wild-type ovaries in the presence of 0.1mM JHB(3) increased ecdysteroid biosynthesis only during the first 18h following eclosion. Female Drosophila melanogaster undergo a pre-vitellogenic reproductive diapause when exposed to low temperature (11 degrees C) and a short-day photoperiod (L12:D12). The rate of ecdysteroid synthesis by the ovaries, but not JH production, increased within 12h of a temperature upshift to 25 degrees C from a basal level of 20+/-1pg/10 pair of ovaries/5h to a sustained level of 150+/-20pg/10 pair/5h. Vitellogenic oocytes were noted in all females within 12h of this temperature upshift. Diapause was also terminated by the injection of 1&mgr;g of 20-hydroxyecdysone into the abdomens of diapausing females as determined by an increase in ovary size, and the appearance of vitellogenic oocytes as compared to controls. These results are consistent with a revised model for the regulation of yolk protein uptake by ovaries in which ecdysteroids, and not JHs, play the prominent role.     

Effects of caffeine on chromosomal loss and nondisjunction in Drosophila melanogaster

Three types of male larvae, normal X males and two types with structurally abnormal X chromosomes (ring X and short X sc ⁴ sc ³, y) were treated during the third instar with 0.5 per cent caffeine in nutrient medium. Upon eclosion, these males were mated to yellow and Oregon-R wild type females. The F₁ generation of each cross was scored for normal (XX and XY) and abnormal (XO and XXY) progeny. Statistical analyses of data demonstrate that caffeine increases chromosomal loss for all genotypes tested. The effect of caffeine on nondisjunction, however, is not clear. There are at least marginal increases in all cases when Oregon-R females are used. Slight increase and decreases noted for offspring of yellow females appear to be dependent upon the genotype of the inseminating male.     

Progressive redistribution of alcohol dehydrogenase during vitellogenesis in Drosophila melanogaster: characterization of ADH-positive bodies in mature oocytes

Summary The use of monoclonal antibodies against Drosophila alcohol dehydrogenase (ADH) provides a powerful tool in the analysis of the tissue and temporal patterns of Adh gene expression. Immunocytochemical techniques at the light- and electron-microscopic levels have been used to determine the distribution of ADH in the ovarian follicles of D. melanogaster during oogenesis. In the early stages of oogenesis, small amounts of ADH are detectable in the cystocytes. At the beginning of vitellogenesis (S₇), ADH appears to be located mainly in the nurse cells. From stage S₉ onwards, the ADH protein is evenly distributed in the ooplasm until the later stages of oogenesis (S_13–14), when multiple ADH-positive bodies of varying size appear in the ooplasm. This change in distribution is a result of the compartmentalization of the ADH protein within the glycogen yolk or -spheres. Yolk becomes enclosed within the lumen of the primitive gut during embryonic development, and thus our results suggest a mechanism for the transfer of maternally-inherited enzymes to the gut lumen via yolk spheres.