Biomedical subjects

S Pimpinelli

Publications and source records attributed to S Pimpinelli.

At least 55 records · Page 3Linked to original sources

PubMed1985

Centromere ultrastructure in germ-line chromosomes of Parascaris.

Ultrastructural analysis of the centromere in germ-line mitotic chromosomes of Parascaris univalens and Parascaris equorum revealed that these chromosomes are holocentric. In thin longitudinal sections of both species the kinetochore appeared as a continuous plate (up to 3.8 micron long) and displayed a layered structure. This structure consisted of electron-dense inner and outer layers (average width 10 nm) separated by a less dense middle layer (25 nm wide), which had transverse electron-dense bars (10 nm wide) regularly spaced every 25-30 nm. Thus the ladderlike kinetochore profile observed in Parascaris gonial mitotic chromosomes represents a different type of organization from that of the classical trilaminar kinetochore found in both holocentric and monocentric chromosomes.

Animals↗

PubMed1985

A cell division mutant of Drosophila with a functionally abnormal spindle.

Normal distribution of chromosomes to daughter cells is insured by the proper functioning of the spindle. Homozygosity for a semi-lethal mutation of Drosophila melanogaster (abnormal spindle) altering this structure has the following effects: the mitotic cycle is arrested in metaphase, leading to a high frequency of polyploid cells; sex chromosome disjunction during male meiosis is severely affected, as revealed by the resulting exceptional (diplo and nullo) gametes (microscopic examination of spermiogenesis confirms this aberrant segregation); meiotic spindles of living cells are morphologically abnormal; and tubulins extracted from mutant larvae are normal in amount, electrophoretic mobility, and ability to form microtubules in vitro. The results suggest that the mutant phenotype is due to an altered structural component of the spindle other than tubulins.

Animals↗

PubMed1985

On biological functions mapping to the heterochromatin of Drosophila melanogaster.

We examined the behavior of an autosomal recessive maternal-effect mutation, abnormal-oocyte (abo), that is located in the euchromatin of the left arm of chromosome 2. When homozygous in females, abo results in a marked reduction in the probability that an egg produced by a mutant mother will develop into an adult. However, this probability is increased if the fertilizing sperm delivers to the egg either a normal allele of the maternal-effect gene or a specific type of heterochromatin (called ABO) that is located in small regions of the X and Y chromosome constitutive heterochromatin as well as in some autosomal heterochromatin. These regions, moreover, all react to Hoechst 33258 fluorescent dye identically and specifically. The amelioration of the maternal effect produced by this heterochromatin differs temporally from that caused by the normal allele of the euchromatic gene: the heterochromatin reduces only precellular blastoderm mortality, whereas the normal allele of the euchromatic gene reduces only postblastoderm mortality. Thus, although the genome of the preblastoderm Drosophila embryo is apparently mostly silent, the ABO-containing heterochromatin functions at this early time. Finally, preliminary data indicate that abo is but one member of a cluster of linked genes, each of which interacts with its own normal allele and with a different, locus-specific, heterochromatic factor. From these observations, it appears that Drosophila heterochromatin contains developmentally important genetic elements, and that a functional concomitant of heterochromatic location is gene action at a developmental stage during which the activity of the euchromatic genome is as yet undetectable. Some general implications of these inferences are considered.

Animals↗

PubMed1981

Cytological dissection of sex chromosome heterochromatin of Drosophila hydei.

Prophase chromosomes of Drosophila hydei were stained with 0.5 microgram/ml Hoechst 33258 and examined under a fluorescence microscope. While autosomal and X chromosome heterochromatin are homogeneously fluorescent, the entirely heterochromatic Y chromosome exhibits an extremely fine longitudinal differentiation, being subdivided into 18 different regions defined by the degree of fluorescence and the presence of constrictions. Thus high resolution Hoechst banding of prophase chromosomes provides a tool comparable to polytene chromosomes for the cytogenetic analysis of the Y chromosome of D. hydei. - D. hydei heterochromatin was further characterized by Hoechst staining of chromosomes exposed to 5-bromodeoxyuridine for one round of DNA replication. After this treatment the pericentromeric autosomal heterochromatin, the X heterochromatin and the Y chromosome exhibit numerous regions of lateral asymmetry. Moreover, while the heterochromatic short arms of the major autosomes show simple lateral asymmetry, the X and the Y heterochromatin exhibit complex patterns of contralateral asymmetry. These observations, coupled with the data on the molecular content of D. hydei heterochromatin, give some insight into the chromosomal organization of highly and moderately repetitive heterochromatic DNA.

Animals↗

PubMed1980

Effects of recombination-deficient and repair-deficient loci on meiotic and mitotic chromosome behavior in Drosophila melanogaster.

The results of recent genetic and cytological studies on recombination-defective and repair-defective mutants of Drosophila melanogaster are summarized. These studies show that there is substantial overlap between the functions used in various aspects of DNA metabolism in Drosophila. Most loci first identified by either recombination-defective or mutagen-sensitive mutants have been shown also to function in nonmutagenized mitotic cells where their action is necessary to maintain the integrity of the genome: mutants at particular loci produce elevated frequencies of chromosome breakage, mitotic exchange, mutation, and/or chromosome loss. Genetic studies of meiotic recombination show that many of the loci identified by recombination-defective mutants restrict where along the chromosome arms exchange may occur. Recent EM studies suggest that the products of at least some of these loci are components of recombination nodules. Region-specific control of DNA metabolism is also indicated by the finding of nonrandom patterns of chromosome breakage in some mutagen-sensitive mutants. Recombination-defective mutants at two loci have been studied for their effects on sister chromatid exchanges (SCEs) and x-ray induced aberrations. Mutants at both loci are defective in steps necessary for the production of symmetrical chromatid interchanges but have little effect on SCEs.

Animals↗

PubMed1980

Intraspecific polymorphism of sex chromosome heterochromatin in two species of the Anopheles gambiae complex.

The Hoechst 33258 banding pattern of the mitotic chromosomes of several laboratory and natural populations of the sibling species A. gambiae and A. arabiensis has been analyzed. A clear intraspecific polymorphism of sex chromosome heterochromatin has been observed. Nevertheless in each species heterochromatic variations fall within a characteristic species-specific pattern. Moreover, while laboratory polulations tend to be monomorphic for a given heterchromatic variant, natural populations exhibit a high degree of intrapopulation polymorphism. The possible role of sex chromosome heterochromatin in controlling fertility and mating behaviour of Anopheles mosquitoes is discussed.

Animals↗

PubMed1980

Relationships among chromatid interchanges, sister chromatid exchanges, and meiotic recombination in Drosophila melanogaster.

Repair- and recombination-defective mutations at two loci (mei-9 and mei-41) of Drosophila melanogaster have been examined for their effects on the induction of chromosome aberrations by x-rays and the formation of sister chromatid exchanges (SCEs). Irradiation of larval neuroblast cells during the S phase with x-rays showed that mutants at both of these loci are about 10 times more sensitive than wild type to the induction of chromosome aberrations. The pattern of induced aberrations was characteristic for each mutant locus: in cells bearing mei-9 mutations most breaks were chromatid deletions, whereas in the presence of mei-41 mutations similar frequencies of chromatid and isochromatid deletions were observed. Furthermore, chromatid interchanges could not be induced in cells carrying mei-9 alleles; therefore these mutations define a step necessary for chromatid rejoining. mei-41 alleles also define a function involved in the formation of chromatid interchanges; total exchanges were less frequent than expected from nonmutant controls; and the proportion of exchanges arising by symmetrical rejoining was markedly reduced. These data indicate that chromatid and isochromatid deletions have different molecular steps in their formation, and that different molecular mechanisms are also involved in the symmetrical and unsymmetrical rejoining in chromatid interchanges. Neuroblast cells of larvae bearing mei-9 and mei-41 alleles were also treated for 13 hr with 5-bromodeoxyuridine at 9 mug/ml in order to differentiate sister chromatids for the scoring of SCEs. Whereas mei-41 had a normal level of SCEs, mei-9 exhibited a frequency of SCEs that was about 70% that of the control. Because both mei-9 and mei-41 mutations result in defective meiotic recombination, these data suggest that they define steps shared by symmetrical interchange formation and meiotic recombination that do not participate in the formation of most SCEs.

Animals↗

PubMed1979

Simultaneous production of Q and R bands after staining with chromomycin A3 or olivomycin.

Human and mouse chromosomes, stained with either chromomycin A3 or olivomycin, which bind preferentially to G - C-rich DNA (where G is guanosine and C is cytosine), exhibit a Q or a reverse banding pattern, depending on the wavelength used for excitation. The two complementary banding patterns can be observed in the same metaphase simply by changing the combination of excitation filters. These data suggest, therefore, that in addition to base composition, other factors are involved in the production of chromosome banding by chromomycin A3 and olivomycin.

Animals↗

PubMed1979

Lack of spontaneous sister chromatid exchanges in somatic cells of Drosophila melanogaster.

Neural ganglia of wild type third-instar larvae of Drosophila melanogaster were incubated for 13 hours at various concentrations of BUdR (1, 3, 9, 27 micrograms/ml). Metaphases were collected with colchicine, stained with Hoechst 33258, and scored under a fluorescence microscope. Metaphases in which the sister chromatids were clearly differentiated were scored for the presence of sister-chromatid exchanges (SCEs). At the lowest concentration of BUdR (1 microgram/ml), no SCEs were observed in either male or female neuroblasts. The SCEs were found at the higher concentrations of BUdR (3, 9, And 27 micrograms/ml) and with a greater frequency in females than in males. Therefore SCEs are not a spontaneous phenomenon in D. melanogaster, but are induced by BUdR incorporated in the DNA. A striking nonrandomness was found in the distribution of SCEs along the chromosomes. More than a third of the SCEs were clustered in the junctions between euchromatin and heterochromatin. The remaining SCEs were preferentially localized within the heterochromatic regions of the X chromosome and the autosomes and primarily on the entirely heterochromatic Y chromosome.--In order to find an alternative way of measuring the frequency of SCEs in the Drosophila neuroblasts, the occurrence of double dicentric rings was studied in two stocks carrying monocentric ring-X chromosomes. One ring chromosome, C(1)TR94--2, shows a rate of dicentric ring formation corresponding to the frequency of SCEs observed in the BUdR-labelled rod chromosomes. The other ring studied, R(1)2, exhibits a frequency of SCEs higher than that observed with both C(1) TR94--2 and rod chromosomes.

Animals↗

PubMed1979

Effects of distamycin A on human leukocytes in vitro.

Distamycin A, an oligopeptide antibiotic, supplied at various concentrations for 24 h to human leukocytes in culture, has induced the appearance on some chromosomes of specific areas lacking spiralization. In particular, the centromeric regions of chromosomes 1, 3 and one C-group chromosome and the distal part of the long arm of the Y chromosome were despiralized. The possible nature of these regions is discussed.

Cells, Cultured↗

PubMed1978

3h-actinomycin-D binding to mitotic chromosomes of Drosophila melanogaster.

The binding of 3H-AMD to the metaphase chromosomes of Drosophila melanogaster has been analyzed after two different periods of exposure to photographic emulsion. The entirely heterochromatic Y chromosome was markedly less labelled than euchromatin and other heterochromatic regions. Moreover, the few grains present on the Y chromosome were clustered in two regions, one localized in the middle Ys and the other in the proximal third of YL. This labelling pattern is not affected by removing histones with a 2-hour treatment with 2N HCl. It is suggested that the specific underlabelling of the Y chromosome reflects a peculiar AT richness.

Animals↗

PubMed1977

Fluorescence banding techniques in the identification of sibling species of the anopheles gambiae complex.

The mitotic chromosomes of the sibling species A and B of the Anopheles gambiae complex were stained with Hoechst 33258 and examined by fluorescence microscopy. The autosomes fluoresce homogenously and similarly in both species while the sex chromosomes differ in the location and brightness of some heterochromatic blocks. These cytochemical differences allow the cytotaxonomic identification of these cryptic species.

Animals↗

PubMed1977

Mutagen specificity in the induction of chromosomal aberrations in somatic cells of Drosophila melanogaster.

The distribution of chromosomal aberrations between and within chromosomes of male D, melanogaster somatic cells after treatment with UV has been analyzed. -- Distribution of the breaks between chromosomes was largely nonrandom since we found a higher aberration frequency than that expected on the Y chromosome. Moreover, within the chromosomes the aberrations are clustered in the pericentromeric heterochromatic regions. The above distribution is compared with that of the breaks induced by X rays and methyl-methane-sulphonate (MMS) which were distributed in a different pattern.

Animals↗

PubMed1976

Characterization of Drosophila heterochromatin. I. Staining and decondensation with Hoechst 33258 and quinacrine.

A number of preliminary experiments have shown that the fluorescence pattern of Hoechst 33258, as opposed to that of quinacrine, varies with the concentration of dye. The metaphase chromosomes of D. melanogaster, D. simulans, D. virilis, D. texana, D. hydei and D. ezoana have therefore been stained with two concentrations of H 33258 (0.05 and 0.5 mug/ml in phosphate buffer at pH 7) and with a single concentration of quinacrine (0.5% in absolute alcohol). The three fluorescence patterns so obtained were shown to be somewhat different in some of the species and the coincide in others. All three stainings gave an excellent longitudinal differentiation of heterochromatin while euchromatin fluoresced homogeneously. Living ganglion cells of the six species mentioned above were treated with quinacrine and H 33258. Quinacrine induced a generalized lengthening and swelling of the chromosomes and H 33258 the decondensation of specific heterochromatic regions. A correlation of the base composition of the satellite DNAs contained in the heterochromatin of the species studied with the relative fluorescence and decondensation patterns showed that: 1) the extremely fluorochrome bright areas and those decondensed are present only in species containing AT rich satellite DNA; 2) the opposite is not true since some AT-rich satellite DNAs are neither fluorochrome bright nor decondensed; 3) there is no good correspondence between Hoechst bright areas and the decondensed ones. AT richness therefore appears to be a necessary but not sufficient condition both for bright fluorescence and decondensation. Some cytological evidence suggests that similarly AT rich satellite DNAs respond differently in fluorescence and decondensation because they are bound to different chromosomal proteins. A combination of the results of fluorescence and decondensation revealed at least 14 types of heterochromatin; 4-7 of which are simultaneously present in the same species. Since closely related species (i.e. D. melanogaster and D. simulans; D. virilis and D. texana) show marked differences in the heterochromatic types they contain, it can be suggested that within the genus Drosophila qualitative variations of heterochromatin have played an important role in speciation.

Animals↗

PubMed1976

Characterization of Drosophila heterochromatin. II. C- and N-banding.

The C- and N-banding patterns of D. melanogaster, D. simulans, D. virilis, D. texana, D. ezoana and D. hydei were studied in comparison with quinacrine and Hoechst banding patterns. In all these Drosophila species the C bands correspond to the heterochromatin as revealed by the positive heteropycnosis in the prometaphase chromosomes. The N bands have the following characteristics: 1) they are always localized on the heterochromatin and generally do not correspond to the C bands; 2) they do not correspond to the nucleolar organizing regions; 3) they are inversely correlated with fluorescence, i.e., they correspond to regions which are scarcely, if at all, fluorescent after Hoechst 33258 or quinacrine staining; 4) they are localized both on regions containing AT rich satellite DNA and on those containing GC rich satellite DNA.

Animals↗

PubMed1976

Effect of Hoechst 33258 on Chinese hamster chromosomes.

Cells of the Chinese hamster strain C-125 were treated for different time intervals with H 33258, a bibenzimidazole derivative. The same compound was used to stain fixed cells of the same strain. H 33258 induced in cells in culture specific areas of reduced spiralization on the metaphase chromosomes of some cells. These probably correspond to DNA segments rich in A-T bases interspersed along the chromosomes. Probably H 33258 acts during S period of cell cycle. The banding obtained by staining with H 33258 is similar to that induced by quinacrine dihydrochloride but shows a better resolution.

Adenine Nucleotides↗

PubMed1976

X-ray induction of chromatid interchanges is somatic cells of Drosophila melanogaster: variations through the cell cycle of the pattern of rejoining.

The relative frequencies of X-ray-induced symmetrical and asymmetrical chromatid interchanges were analysed as a function of the cell cycle in somatic cells of Drosophila melanogaster. In exchanges between autosomes, during the S phase, a clear prevalence of asymmetrical rejoining was observed. Later, this prevalence became reduced as the cell approached mitosis. On the other hand, in the exchanges between X-chromosomes in the females there were no significant variations in the type of rejoining through the cell cycle, there being a slight but consistent preference for symmetrical rejoining. These data are discussed in relation to the hypothesis proposed by Olivieri et al. [12] on the organization of the chromosomal sub-units in the interphase nucleus.

Animals↗

PubMed1976

Effects of Hoechst 33258 on human leukocytes in vitro.

The benzimidazole derivative Hoechst 33258 was added at various concentrations to human leukocyte cultures. After 16 or 24 h of treatment, with concentrations equal to or greater than 100 mug/ml of Hoechst 33258, a number of chromosomes showed regions in which the chromatin was undercontracted. The centromeric regions of chromosome 1 and, more rarely, of chromosomes 3 and 9 appeared to be decondensed. Short decondensed regions were also present on the long arms of chromosomes 1 and 2. The possible nature of these regions is discussed.

Benzimidazoles↗