Heterochromatin variation in chromosome X in a natural population of Anopheles willmori (Diptera: Culicidae) of Thailand

Among the eight families of Anopheles willmori derived from individual wild-caught females collected from Chiangmai Province (northern Thailand) and examined, four isofemale lines showed variation in the X chromosome, including the normal X₁ and three aberrant types (X₃, X₄ and X_L). It is postulated that these different types of X chromosomes could have arisen as a result of spontaneous chromosomal rearrangements involving tandem translocation and paracentric inversion followed by acquisition of constitutive heterochromatin. Such rare events of chromosomal changes have become established in the natural population of An. willmori in northern Thailand.     

Cytological identification of two X-chromosome types in the wood lemming (Myopus schisticolor)

In the wood lemming (Myopus schisticolor) three genetic types of sex chromosome constitution in females are postulated: XX, X^*X and X^*Y (X^*=X with a mutation inactivating the male determining effect of the Y chromosome). Males are all XY. It is shown in the present paper that the two types of X chromosomes, X and X^*, exhibit differences in the G-band patterns of their short arms. In addition, it was demonstrated in unbanded chromosomes that the short arm in X^* is shorter than in X. The origin of these differences is still obscure; but they allow to identify and to distinguish the individual types of sex chromosome constitution, as of XX versus X^*X females and of X^*Y females versus XY males, on the basis of G-banded chromosome preparations from somatic cells.     

Late replication patterns in adult and embryonic mice carrying Searle's X-autosome translocation

Bromodeoxyuridine-dye technique analysis of X chromosome DNA synthesis in female adult and fetal mice carrying the balanced form of the T(X; 16) 16H translocation demonstrated that the structurally normal X chromosome was late replicating (and hence presumably inactive) in 93% of the adult cells and 99% of the 9-day embryo cells, with the X¹⁶ chromosome late replicating in the remaining cells. We conclude from these results that in T16H/+ females either there is preferential inactivation of the normal X chromosome or that, if inactivation is random, cell selection takes place before 9 days of development. Two 9-day female embryos with an unbalanced karyotype were also studied; both had two late-replicating chromosomes in most of their cells, one being the chromosome 16^X, the other a normal X chromosome. These results, together with the presence of a late-replicating X¹⁶ chromosome in T16H/+ adult and fetal mice, support the concept that more than one inactivation center is present on the X chromosome of the mouse because the X¹⁶ and the 16^x chromosomes can be late replicating.     

Properties of the sex chromosomes of Lucilia cuprina deduced from radiation studies

From a study of radiation-induced X-chromosome deletions the locus of black body (b) has been localized to the proximal portion of C-band defined euchromatin. Radiation produced mostly X-chromosome deletions rather than point mutations, total X or Y chromosome loss through breakage, or increased frequency of non-disjunction. Aberrant sex ratios obtained indicate that the X chromosome carries vital loci that were deleted with b ⁺ in many cases. The X/O karyotype produces fertile adult females with a characteristic phenotype which is also produced by X deletions. Sex chromosome non-disjunction to give X/O females and X/X/Y males is normally rare but is enhanced by the presence of chromosome rearrangements even when the X and Y are not involved.     

A transmissible dicentric chromosome in Drosophila melanogaster

A transmissible dicentric chromosome was recovered in Drosophila melanogaster. The radiation-induced secondary chromosome rearrangement consists essentially of the entire Y and fourth chromosomes joined by 2R heterochromatin. The Y ^S · Y ^L 2Rh4 · chromosome pairs with the X and the free fourth chromosome to form a trivalent in meiosis that is unusual because it forms few chromosome bridges in primary spermatocytes and is transmitted at high frequency. We suggest that the orientation of the weaker fourth chromosome kinetochore eventually fails when opposing the stronger Y kinetochore so that the Y ^S · Y ^L 2Rh4 · moves to the pole to which the Y kinetochore is oriented. There is however an increased frequency of sex chromosome nondisjunction (14%) and of chromosome laggards (6%) in primary spermatocytes; the frequency of exceptional progeny of males containing the Y ^S · Y ^L 2Rh4 · was 7.44% compared with 0.25% in the controls. Disruption of normal sex chromosome disjunction also occurs in females containing the Y ^S · Y ^L 2Rh4 · and a compound X chromosome; the frequency of exceptional progeny was 2.55% versus 0.91% in the controls. Chromosome nondisjunction appears to occur when orientation of the X and Y kinetochores to the same pole is stabilized through tension by the orientation of one or both fourth chromosome kinetochores to the opposite pole. During anaphase, the orientation of the fourth chromosome kinetochore of the Y ^S · Y ^L 2Rh4 · appears to fail and the X and Y ^S · Y ^L 2Rh4 · chromosomes move to the same pole. Y ^S · Y ^L 2Rh4 · chromosome laggards occur with both the Y and fourth chromosome kinetochores amphitelically oriented. This orientation appears to be stable as a result of equal opposing forces toward opposite poles.     

An X‐linked Myh11‐CreERT2 mouse line resulting from Y to X chromosome‐translocation of the Cre allele

The Myh11‐CreER^T2 mouse line (Cre⁺) has gained increasing application because of its high lineage specificity relative to other Cre drivers targeting smooth muscle cells (SMCs). This Cre allele, however, was initially inserted into the Y chromosome (X/Y^Cre+), which excluded its application in female mice. Our group established a Cre⁺ colony from male ancestors. Surprisingly, genotype screening identified female carriers that stably transmitted the Cre allele to the following generations. Crossbreeding experiments revealed a pattern of X‐linked inheritance for the transgene (k > 1000), indicating that these female carries acquired the Cre allele through a mechanism of Y to X chromosome translocation. Further characterization demonstrated that in hemizygous X/X^Cre+ mice Cre activity was restricted to a subset arterial SMCs, with Cre expression in arteries decreased by 50% compared to X/Y^Cre+ mice. This mosaicism, however, diminished in homozygous X^Cre+/X^Cre+ mice. In a model of aortic aneurysm induced by a SMC‐specific Tgfbr1 deletion, the homozygous X^Cre+/X^Cre+ Cre driver unmasked the aortic phenotype that is otherwise subclinical when driven by the hemizygous X/X^Cre+ Cre line. In conclusion, the Cre allele carried by this female mouse line is located on the X chromosome and subjected to X‐inactivation. The homozygous X^Cre+/X^Cre+ mice produce uniform Cre activity in arterial SMCs.     

Conserved autonomy of replication of the X-chromosomes in hybrids of Drosophila miranda and Drosophila persimilis

The chromosome complement of hybrid males from the cross between Drosophila miranda female and D. persimilis male provides an interesting chromosomal situation where an autosome, the 3rd chromosome of D. persimilis, coexists with a homologue that developed into a sex chromosome, the X₂ in D. miranda. Except for certain inversions and a few minor translocations, these two chromosomes (X₂ and the 3rd) still look alike as polytene elements. However, in hybrid males pairing of the two chromosomes, the X₂ and 3rd, is rare, while in female hybrids it occurs frequently. — ³H-TdR labeling shows that while the X₂ and 3rd chromosomes replicate synchronously in hybrid female, in the hybrid male the former completes its replication earlier than the 3rd chromosome, as do the two arms of the X₁ (XL and XR). The frequency and relative intensity of ³H-TdR labeling of each site of the X₂ and that of the 3rd chromosome in hybrid males closely agree with those of the corresponding sites in the X₂ of the miranda male and the 3rd chromosome of the persimilis male (or female), respectively. The results suggest that timing and rate of replication of the X₂ are determined autonomously and follow the pattern in the respective parental species.     

<Emphasis Type="Italic">Drosophila simulans Lethal hybrid rescue</Emphasis> mutation (<Emphasis Type="Italic">Lhr</Emphasis>) rescues inviable hybrids by restoring X chromosomal dosage compensation and causes fluctuating asymmetry of development

The Drosophila simulans Lhr rescues lethal hybrids from the cross of D. melanogaster and D. simulans. We describe here, the phenotypes of Lhr dependent rescue hybrids and demonstrate the effects of Lhr on functional morphology of the salivary chromosomes in the hybrids. Our results reveal that the phenotypes of the ‘Lhr dependent rescued’ hybrids were largely dependent on the genetic background and the dominance in species and hybrids, and not on Lhr. Cytological examination reveal that while the salivary chromosome of ‘larval lethal’ male carrying melanogaster X chromosome was unusually thin and contracted, in ‘rescued’ hybrid males (C _mel X _mel Y _sim ; A _mel A _sim ) the X chromosome showed typical pale staining, enlarged diameter and incorporated higher rate of ³H-uridine in presence of one dose Lhr in the genome. In hybrid males carrying simulans X chromosome (C _mel X _sim Y _mel ; A _mel A _sim ), enlarged width of the polytene X chromosome was noted in most of the nuclei, in Lhr background, and transcribed at higher rate than that of the single X chromosome of male. In hybrid females (both viable, e.g., C _mel X _mel X _sim ; A _mel A _sim and rescued, e.g., C _mel X _mel X _mel ; A _mel A _sim ), the functional morphology of the X chromosomes were comparable to that of diploid autosomes in presence of one dose of Lhr. In hybrid metafemales, (C _mel X _mel X _mel X _sim ; A _mel A _sim ), two dose of melanogaster X chromosomes and one dose of simulans X chromosome were transcribed almost at ‘female’ rate in hybrid genetic background in presence of one dose of Lhr. In rescued hybrid males, the melanogaster-derived X chromosome appeared to complete its replication faster than autosomes. These results together have been interpreted to have suggested that Lhr suppresses the lethality of hybrids by regulating functional activities of the X chromosome(s) for dosage compensation.     

Increased 5-HT3-mediated signalling in pelvic afferent neurons from mice deficient in P2X2 and/or P2X3 receptor subunits

Extracellular ATP and 5-hydroxytryptamine (5-HT) are both involved in visceral sensory pathways by interacting with P2X and 5-HT₃ receptors, respectively. We have investigated the changes in P2X and 5-HT₃-mediated signalling in pelvic afferent neurons in mice deficient in P2X₂ and/or P2X₃ subunits by whole-cell recording of L₆–S₂ dorsal root ganglion (DRG) neurons and by multi-unit recording of pelvic afferents of the colorectum. In wildtype DRG neurons, ATP evoked transient, sustained or mixed (biphasic) inward currents. Transient currents were absent in P2X₃ ^−/− neurons, whereas sustained currents were absent in P2X₂ ^−/− DRG neurons. Neither transient nor sustained currents were observed following application of ATP or α,β-methylene ATP (α,β-meATP) in P2X₂/P2X₃ ^Dbl−/− DRG neurons. 5-HT was found to induce a fast inward current in 63% of DRG neurons from wildtype mice, which was blocked by tropisetron, a 5-HT₃ receptor antagonist. The percentage of DRG neurons responding to 5-HT was significantly increased in P2X ₂ ^−/−, P2X₃ ^−/− and P2X₂/P2X₃ ^Dbl−/− mice, and the amplitude of 5-HT response was significantly increased in P2X₂/P2X₃ ^Dbl−/− mice. The pelvic afferent response to colorectal distension was attenuated in P2X₂/P2X₃ ^Dbl−/− mice, but the response to serosal application of 5-HT was enhanced. Furthermore, tropisetron resulted in a greater reduction in pelvic afferent responses to colorectal distension in the P2X₂/P2X₃ ^Dbl−/− preparations. These data suggest that P2X receptors containing the P2X₂ and/or P2X₃ subunits mediate purinergic activation of colorectal afferents and that 5-HT signalling in pelvic afferent neurons is up-regulated in mice lacking P2X₂ or P2X₃ receptor genes. This effect is more pronounced when both subunits are absent.     

Primary and secondary nonrandom X chromosome inactivation in early female mouse embryos carrying Searle's translocation T(X; 16)16H

By means of a cytological technique involving 5-bromodeoxyuridine, acridine orange, and fluorescence microscopy, the asynchronously replicating, hence genetically inactivated, X chromosome was identified in 6-to 8-day embryos from female mice heterozygous for Searle's translocation T(X;16)16H (abbreviated as T16H) mated with either karyotypically normal males or males carrying Cattanach's translocation T(X;7)lCt in order to analyse the way in which the total inactivation of the normal X is achieved in adult T16H heterozygotes. Embryos examined included 9 Xⁿ/X⁽⁷⁾;16/16, 3X¹⁶/Xⁿ;16^x/16, 12X¹⁶/X⁽⁷⁾;16^x/16, 5 X¹⁶/Xⁿ; 16/16, 8 X¹⁶/X⁽⁷⁾; 16/16 and 2 Xⁿ/Y; 16^x/16/16. In these notations X¹⁶, 16^x, X⁽⁷⁾ and Xⁿ represent Searle's X with the centromeric segment of the X, Searle's X with the centomeric segment of chromosome 16, Cattanachs's X with insertion of a chromosome 7 segment, and normal X, respectively. The X⁽⁷⁾ exerted no apparent effect upon embryonic development up to the 8th day of gestation and X chromosome inactivation. — The asynchronously replicating X was the Xⁿ in X¹⁶/ Xⁿ;16^x/16 and X⁽⁷⁾ in X¹⁶/X⁽⁷⁾;16^x/16 embryos except a small number of cells on day 6 (13/493) and on day 7 (1/886) in which almost the entire 16^x replicated asynchronously. The X¹⁶, on the other hand, never showed replication asynchrony. That the X¹⁶ is indeed unable to become inactivated was indicated by the observation that the X¹⁶ as well as Xⁿ or X⁽⁷⁾ did not replicate asynchronously in Xⁿ/X¹⁶; 16/16 and X¹⁶/X⁽⁷⁾; 16/16 embryos. X¹⁶-inactive cell lines, if occurring, should have been genetically less unbalanced than any other cell line in such embryos. It is highly likely therefore that the ultimate inactivation pattern in T16H heterozygotes has been accomplished by (1) the inability of the X¹⁶ to become inactive; (2) inactivation in favor of the Xⁿ; and (3) rapid elimination of 16^x-inactive cells. Severe growth retardation and early death of X¹⁶/Xⁿ;16/16 and X¹⁶/X⁽⁷⁾; 16/16 embryos having no inactive X suggested that functional X disorny is detrimental to embryogenesis. These embryos further indicated that the concurrence of at least two X chromosomal loci separated by the T16H breakpoint is necessary for the homologous X chromosome becoming inactivated.     

Univalent sex chromosomes in spermatocytes of Sxr-carrying mice

Pachytene configurations of the sex chromosomes were studied in whole-mount, silver-stained preparations of spermatocytes in mice with XY,Sxr, XX,Sxr, XO,Sxr, XO,Sxr+5¹² and T(X;4)37H,YSxr chromosomes, and non-Sxr-carrying controls. XY,Sxr males showed an increased number of X and Y univalents and of self-synapsed Y chromosomes. In T(X;4)37H,YSxr males an increased proportion of trivalent+Y configurations was also accompanied by higher numbers of self-paired Y univalents; the proportion of trivalent+X⁴ was not increased, but that of self-synapsed X⁴ univalents was. There was more selfsynapsis in cells containing one univalent than in cells containing two univalents. Spermatocytes of XX,Sxr mice contained single univalent X, which was never seen to be self-synapsed, but self-synapsis of the X occurred in a proportion of cells in XO,Sxr males. There were no self-paired X chromosomes in the XO,Sxr+5¹² mouse although lowlevel pairing of the 5¹² chromosome occurred. All four XX,Sxr and XO,Sxr males contained testicular sperm, and testicular sperm were also present in one T(X;4)37H male, while another such male had sperm in the caput. It is concluded that (1) self-synapsis of univalents is affected by variable conditions in the cell as well as by the DNA sequences of the chromosome, and (2) that the level of achievable spermatogenesis is not always rigidly predetermined by a chromosome anomaly but can be modulated by the genetic background.     

Cytogenetics of ticks (Acari: Ixodoidea)

Summary Prior to this paper there have been no reports of a multiple sex chromosome mechanism operative in any tick. The present paper deals with two species of Ixodidae, Amblyomma moreliae and Amblyomma limbatum that exhibit an X₁X₁X₂X₂:X₁X₂Y type of sex chromosome mechanism. Cells from males of both species show nine bivalents plus one sex trivalent. Eleven bivalents were observed in one female A. moreliae. The sex trivalent probably evolved through reciprocal translocation from a system that included ten autosomal bivalents and one sex univalent (the system found in most ixodid species). As a result of the translocation, there are now two X chromosomes (X₁ and X₂) segregating from an unaltered autosome, the neo-Y. A large X chromosome is characteristic of many ticks; in this instance the reciprocal translocation did not change appreciably its relative size.The opinions and assertions contained herein are the private ones of the author and are not to be construed as official or reflecting the views of the Navy Department or the Naval service at large.This study was begun during the tenure of a North Alantic Treaty Organization (National Science Foundation) Postdoctoral Fellowship.     

Chromosome banding in amphibia

A cytogenetic study performed on a population of the South American leptodactylid frog Eleutherodactylus maussi revealed multiple sex chromosomes of the X₁X₁X₂X₂/X₁X₂Y (=XXAA/XXA^Y) type. The diploid chromosome number is 2n=36 in all females and 2n=35 in most males. The multiple sex chromosomes originated by a centric fusion between the original Y chromosome and a large autosome. In male meiosis the X₁X₂Y (=XXA^Y) multiple sex chromosomes form a classical trivalent configuration. E. maussi is the first species discovered in the class Amphibia that is distinguished by a system of multiple sex chromosomes. Only one single male was found in the population with 2n=36 chromosomes and lacking the Y-autosomal fusion. This karyotype (XYAA) is interpreted as the ancestral condition, preceding the occurrence of the Y-autosome fusion.by H.C. Macgregor     

A complex sex-chromosome system in the hare-wallaby Lagorchestes conspicillatus Gould

Among specimens of the spectacled hare-wallaby Lagorchestes conspicillatus Gould (Marsupialia, family Macropodidae) 4 males had 15 chromosomes and 2 females 16 chromosomes. The sex chromosomes are X₁X₁X₂X₂ in the female and X₁X₂Y in the male, the Y being metacentric and both X chromosomes are acrocentric. In about 96% of sperm mother cells at meiosis the sex chromosomes form a chain trivalent and in more than 99% of these this orients convergently so that the X₁ and X₂ move to the same pole. Evidence is presented that L. conspicillatus has evolved from a form with 22 chromosomes including a small X and a minute Y. Autoradiographic studies show that the proximal fifth of the X₁ chromosome replicates late. This is probably the ancestral X chromosome which has been translocated to an autosome. The fate of the original Y is obscure but an hypothesis is proposed that it forms the centromeric region of the Y. A single male had 14 chromosomes and was heterozygous for a translocation involving the centric fusion of two acrocentric autosomes. In about 30% of sperm mother cells the autosomal trivalent did not disjoin regularly but, despite this, all secondary spermatocytes observed at metaphase 2 had balanced complements of chromosomes. It is assumed that unbalanced secondary spermatocytes died before reaching metaphase.     

An inducible change in state on the chromosomes of Sciara: Its effects on the genetic components of the X

Normally in Sciara males the sex chromosome constitution of the soma (X ^m O) differs from that of the germ line (X ^m X ^p ), the single X being of maternal origin. Males which are patroclinous for their sex-linked genes (X ^p O) are found regularly among the progeny of females heterozygous for any one of a number of X-translocations (X X ^t ). The patroclinous males are almost invariably completely sterile. In the present study a comparison is made of the patroclinous males derived from 5 different X-translocations in S. coprophila. The study includes data on the morphology and sexual behavior of the males as well as the cytology of the testis from late 4th instar through pupal life. The study is oriented towards the basic question of whether the paternally-derived X — normally destined to be eliminated — can function properly when it is retained. The comparative study supports our original suggestion that the patroclinous males arise from no-X eggs following 31 disjunction during oogenesis.Dedicated to Professor J. Seiler on the occasion of his 80th birthday.The studies reported here were supported by the National Science Foundation, grants GB-42 and GB 2857, and in part by Contract No. AT-(40-1)-2690 under the Division of Biology and Medicine, U. S. Atomic Energy Commission.     

A Drosophila melanogaster cell line tested for the presence of active NORs by silver staining

Silver staining was used to detect active NORs in a Drosophila melanogaster cell line (C1 82) characterized by dimorphic X chromosomes (XX_L), one of the two Xs showing a marked increase in heterochromatin where the nucleolar organizer (NO) is located. The Q-banding technique was used to determine the karyotype characteristics of the line. Ag-positive NORs appeared only on structurally changed X chromosomes (X_L), both in diploid and tetraploid cells, indicating that rRNA genes of X_L are more active or numerous than those on normal homologues. A possible relationship between NOR stainability, the presence of an increased heterochromatic portion and the selective advantage of XX_L cells, recurrent in numerous Drosophila female lines, is discussed.     

Sex determination and phenotype in wood lemmings with XXY and related karyotypic anomalies

Summary The wood lemming, Myopus schisticolor, possesses a unique sex determining system comprising both XX and XY females. Normal female development in the presence of XY is guaranteed by a mutation on the X, apparently associated with a structural rearrangement in Xp. This mutation inactivates the testis-inducing and male-determining factor on the Y and distinguishes X^* from X, and X^*Y females from XY males. Normal fertility of X^*Y females is ensured by a mitotic (double) nondisjunction mechanism which, at an early fetal stage, eliminates the Y from the germ line and replaces it by a copy of the X^*.Numerical sex chromosome aberrations are not infrequent and the trisomics XXY and X^*XY are relatively common. XXY individuals are sterile males with severe suppression of spermatogenesis. Among X^*XY animals, both males and females, as well as a true lateral hermaphrodite have been observed. Primary deficiency of germ cells, impairment of spermatogenesis and sterility are characteristic traits of the X^*XY males, whereas X^*XY females have normal oogenesis and are fertile. Both these extremes (except female fertility) coexist in the true hermaphrodite described in the present study. These apparently contradictory observations are explainable under the assumption that X^* and X in X^*XY individuals are inactivated non-randomly or that the cells are distributed unequally. Inactivation of the X or X^* determines whether or not the H-Y antigen will be expressed. When comparing conditions in Myopus and in man, an additional assumption has to be made in relation to the gene(s) involved in sex determination, located in Xp:In Myopus they do not escape inactivation, whereas in man they have been claimed to remain active.     

Chromosome mapping of ribosomal genes and histone H4 in the genus Radacridium (Romaleidae)

In this study, two species of Romaleidae grasshoppers, Radacridium mariajoseae and R.nordestinum, were analyzed after CMA₃/DA/DAPI sequential staining and fluorescence in situ hybridization (FISH) to determine the location of the 18S and 5S rDNA and histone H4 genes. Both species presented karyotypes composed of 2n = 23, X0 with exclusively acrocentric chromosomes. CMA₃⁺ blocks were detected after CMA₃/DA/DAPI staining in only one medium size autosome bivalent and in the X chromosome in R. mariajoseae. On the other hand, all chromosomes, except the L1 bivalent, of R. nordestinum presented CMA₃⁺ blocks. FISH analysis showed that the 18S genes are restricted to the X chromosome in R. mariajoseae, whereas these genes were located in the L₂, S₉ and S₁₀ autosomes in R. nordestinum. In R. mariajoseae, the 5S rDNA sites were localized in the in L₁ and L₂ bivalents and in the X chromosome. In R. nordestinum, the 5S genes were located in the L₂, L₃, M₄ and M₅ pairs. In both species the histone H4 genes were present in a medium size bivalent. Together, these data evidence a great variability of chromosome markers and show that the 18S and 5S ribosomal genes are dispersed in the Radacridium genome without a significant correlation.