Comportamento dos cromossômios na meiose de Euryophthalmus rufipennis Laporte (Hemiptera Pyrrhocoridae)

In this paper an account is given of the principal facts observer in the meiosis of Euryophthalmus rufipennis Laporte which afford some evidence in favour of the view held by the present writer in earlier publications regarding the existence of two terminal kinetochores in Hem ip ter an chromosomes as well as the transverse division of the chromosomes. Spermatogonial mitosis - From the beginning of prophase until metaphase nothing worthy of special reference was observed. At anaphase, on the contrary, the behavior of the chromosomes deserves our best attention. Indeed, the chromoso- mes, as soon as they begin to move, they show both ends pronouncedly turned toward the poles to which they are connected by chromosomal fibres. So a premature and remarkable bending of the chromosomes not yet found in any other species of Hemiptera and even of Homoptera points strongly to terminally localized kinetochores. The explanation proposed by HUGHES-SCHRADER and RIS for Nautococcus and by RIS for Tamalia, whose chromosomes first become bent late in anaphase do not apply to chromosomes which initiate anaphase movement already turned toward the corresponding pole. In the other hand, the variety of positions assumed by the anaphase chromosomes of Euryophthalmus with regard to one another speaks conclusively against the idea of diffuse spindle attachments. First meiotic division - Corresponding to the beginning of the story of the primary spermatocytes cells are found with the nucleus entirelly filled with leptonema threads. Nuclei with thin and thick threads have been considered as being in the zygotente phase. At the pachytene stage the bivalents are formed by two parallel strands clearly separated by a narrow space. The preceding phases differ in nothing from the corresponding orthodox ones, pairing being undoubtedly of the parasynaptic type. Formation of tetrads - When the nuclei coming from the diffuse stage can be again understood the chromosomes reappear as thick threads formed by two filaments intimately united except for a short median segment. Becoming progressively shorter and thicker the bivalents sometimes unite their extremities forming ring-shaped figures. Generally, however, this does not happen and the bivalents give origin to more or less condensed characteristic Hemipteran tetrads, bent at the weak median region. The lateral duplicity of the tetrads is evident. At metaphase the tetrads are still bent and are connected with both poles by their ends. The ring-shaped diakinesis tetrads open themselves out before metaphase, showing in this way that were not chiasmata that held their ends together. Anaphase proceeds as expected. If we consider the median region of the tetrads as being terminalized chiasmata, then the chromosomes are provided with a single terminal kinetochore. But this it not the case. A critical analysis of the story of the bivalents before and after the diffuse stage points to the conclusion that they are continuous throughout their whole length. Thence the chromosomes are considered as having a kinetochore at each end. Orientation - There are some evidences that Hemipteran chromosomes are connected by chiasmata. If this is true, the orientation of the tetrads may be understood in the following manner: Chiasmata being hindered to scape by the terminal kinetochores accumulate at the ends of the tetrads, where condensation begins. Repulsion at the centric ends being prevented by chiasmata the tetrads orient themselves as if they were provided with a single kinetochore at each extremity, taking a position parallelly to the spindle axis. Anaphase separation - Anaphase separation is consequently due to a transverse division of the chromosomes. Telophase and secund meiotic division - At telophase the kinetochore repeli one another following the moving apart of the centosomes, the chiasmata slip toward the acentric extremities and the chromosomes rotate in order to arrange themselves parallelly to the axis of the new spindle. Separation is therefore throughout the pairing plane. Origin of the dicentricity of the chromosomes - Dicentricity of the chromosomes is ascribed to the division of the kinetochore of the chromosomes reaching the poles followed by separation and distension of the chromatids which remain fused at the acentric ends giving thus origin to terminally dicentric iso-chromosomes. Thence, the transverse division of the chromosomes, that is, a division through a plane perpendicular to the plane of pairing, actually corresponds to a longitudinal division realized in the preceding generation. Inactive and active kinetochores - Chromosomes carrying inactive kinetochore is not capable of orientation and active anaphasic movements. The heterochromosome of Diactor bilineatus in the division of the secondary spermatocytes is justly in this case, standing without fibrilar connection with the poles anywhere in the cell, while the autosomes are moving regularly. The heterochromosome of Euryophthalmus, on the contrary, having its kinetochores perfectly active ,is correctly oriented in the plane of the equator together with the autosomes and shows terminal chromosomal connection with both poles. Being attracted with equal strength by two opposite poles it cannot decide to the one way or the other remaining motionless in the equator until some secondary causes (as for instances a slight functional difference between the kinetochores) intervene to break the state of equilibrium. When Yiothing interferes to aide the heterochromosome in choosing its way it distends itself between the autosomal plates forming a fusiform bridge which sometimes finishes by being broken. Ordinarily, however, the bulky part of the heterochromosome passes to one pole. Spindle fibers and kinetic activity of chromosomal fragments - The kinetochore is considered as the unique part of the chromosome capable of being influenced by other kinetochore or by the poles. Under such influence the kinetochore would be stimulated or activited and would elaborate a sort of impulse which would run toward the ends. In this respect the chromosome may be compared to a neüròn, the cell being represented by the kinetochore and the axon by the body of the chromosome. Due to the action of the kinetochore the entire chromosome becomes also activated for performing its kinetic function. Nothing is known at present about the nature of this activation. We can however assume that some active chemical substance like those produced by the neuron and transferred to the effector passes from the kinetochore to the body of the chromosome runing down to the ends. And, like an axon which continues to transmit an impulse after the stimulating agent has suspended its action, so may the chromosome show some residual kinetic activity even after having lost its kinetochore. This is another explanation for the kinetic behavior of acentric chromosomal fragmehs. In the orthodox monocentric chromosomes the kinetic activity is greater at the kinetochore, that is, at the place of origin of the active substance than at any other place. In chromosomes provided with a kinetochore at each end the entire body may become active enough to produce chromosomal fibers. This is probably due to a more or less uniform distribution and concentration of the active substance coming simultaneously from both extremities of the chromosome.

Uma nova modalidade de sexo-determinação no grilo sul-americano eneoptera surinamensis

The male of Eneoptera surinamensis (Orthoptera-Eneopteridae) is provided with 9 chromosomes, that is, with 3 pairs of autosomes and 3 sex chromosomes. Spermatogonia. - The autosomes of the spermatogonia are of the same size and U-shaped. One of the sex chromosomes approximately equalling the autosomes in size is telocentric, while the other two are much larger and V-shaped. One of the latter is smaller than the other. The sex chromosomes as showed in Figs. 1 and 2 are designated by X, Yl and Y2, X being the larger V, Yl the smaller one and Y2 the rod-shaped. Primary spermatocytes. - Before the growth period of the spermatocytes all the three sex chromosomes are visible in a state of strong heteropycnosis. X is remarkable in this stage in having two long arms well separated by a wide commissural segment. (Figs. 4, 5 and 6). During the growth period Y2 disappears, while X and Yl remain in a condensed form until metaphase. These may be separated from one another or united in the most varied and irregular manner. (Fig. 7 to 12). In the latter case the segments in contact seem to be always different so that we cannot recognize any homology of parts in the sense os genetics. At diplotene Y2 reappears together with the autosomal tetrads. X and Yl may again be seen as separate or united elements. (Figs. 13 and 14). At later diakinesis and metaphase the three sex chromosomes are always independent from each other, Y2 being typically rod-shaped, X and Yl V-shaped, X being a little larger than Yl. (Fig. 15 to 18). At metaphase the three condensed tetrads go to the equatorial plane, while the sex chromosomes occupy any position at both sides of this plane. In almost all figures which could be perfectly analysed X appeared at one side of the autosomal plate an Yl together with Y2 far apart at the other side. (Figs. 16 and 18). Only a few exception have been found. (Figs. 17 and 19). At anaphase X goes in precession to one pole, Yl and Y2 to the other (Figs. 20 and 21). As it is suggested by the few figures in which a localization of the sex chromosomes different from the normal has been observed, the possibility of other types of segregation of these elements cannot be entirely precluded. But, if this does happen, the resulting gametes should be inviable or give inviable zygotes. Early in anaphase autosomes and sex chromosomes divide longitudinally, being maintained united only by the kinetochore. (Figs. 20 and 21). At metaphase the three sex chromosomes seem to show no special repulsion against each other, X being found in the proximity of Yl or Y2 indifferently. At anaphase, however, the evidences in hand point to a stronger repulsion between X on the one side and both Ys on the other, so that in spite of the mutual repulsion of the latter they finish by going to the same pole. Secondary spermatocytes. - At telophase of the primary spermatocytes all the chromosomes enter into distension without disappearing of view. A nuclear membrane is formed around the chromosomes. All the chromosomes excepting Y2 which has two arms, are four-branched. (Fig. 22). Soon the chromosomes enter again into contraction giving rise to the secondary metaphase plate. Secondary spermatocytes provided as expected with four and five chromosomes are abundantly found. (Figs. 23 and 24). In the former all chromosomes are X-shaped while in the latter there is one which is V-shaped. This is the rod- shaped Y2. In the anaphase of the spermatocytes with four chromosomes all the chromosomes are V-shaped, one of them (X) being much larger than the others. In those with five there is one rod-shaped chromosome (Y2). (Fig. 25), Spermatids. Two classes of spermatids are produced, one with X and other with Yl and Y2. All the autosomes as well as Y2 soon enter into solution, X remaining visible for long time in one class and Yl in the other. (Figs. 26 and 27). Since both are very alike at this stage, one cannot distinguish the two classes of spermatids. Somatic chromosomes in the famale. - In the follicular cells of the ovary 8 chromosomes were found, two of which are much larger than the rest. (Figs. 29 and 30). These are considered as being sex chromosomes. CONCLUSION: Eneoptera surinamensis has a new type of sex-determining mechanism, the male being X Yl Y2 and the female XX. The sex chromosomes segregate without entering into contact at metaphase or forming group. After a review of the other known cases of complex sex chromosome mechanism the author held that Eneoptera is the unique representative of a true determinate segregation of sex chromosomes. Y2 behaving as sex chromosome and as autosome is considered as representing an intermediary state of the evolution of the sex chromosomes.

Comportamento dos cromossômios no gênero Hypselonotus (Hemiptera -Coreidae)

The three species studied have 19 chromosomes, being one heterochromosome, one pair of microchromosomes and 8 pairs of autosomes. The microchromosomes of Hypselonotus fulvus are amongst the largest we know. During the synizesis, in Hypselonotus fulvus, we can see in several strands that scape from the chromatic knot a place in which they are widley open. As, in that phase the chromosomes have both ends converging to the same place, the openings suggest a side-to-side pairing of the chromosomal threads. The tetrads are like that studied by Piza (1945-1946). The bivalents are united side by side at their entire length. The unpaired part at the midle of the bivalents gives origin to the arms of the cross-shapede tetrads. The chromosomes have a kinetochore at each end. The bivalents sometimes unite their extremities to form ring-shaped figures, which open themselves out before metaphase. The tetrads are oriented parallelly to the spindle axis. At telophase the kinetochores repeli one another, the chiasmata, if present, slip toward the acentric extremities and the chromosomes rotate in order to arrange themselves parallelly to the axis of the new spindle. Separation is therefore through the pairing plane. In the spermatogonial anaphase of Hypselonotus subterpunctatus the chromosomes are curved to the poles, like those described by PIZA (1946) and PIZA and ZAMITH (1946). The sex chromosomes in Hypselonotus interruptus and Hypselonotus fulvus appears longitudinally divided. It is oriented with the ends in the plane of the equator and its chomatids separate by the plane of division. In the second division the sex chromosome, provided as it is with an actve klnetochore at each end, orients itself with its length parallelly to the spindle axis and passes undivided to one pole. Sometimes it is distended between the poles. This corresponds to case (a) established by PIZA (1946) for the sex chromosomes of Hemiptera In Hypselonotus subterpunctatus the sex chromosome, in the first division of the spermatocytes, orients like the tetrads and divides transversaly. In the second division, as its kinetochore becomes inactive, it remans monocentric, does not orient in the spindle, and is finally enclosed in the nearer nucleus. In the secondary telophase it recuperates its dicentricity like the autosomal chromatids. This behavior corresponds to case (c) of PIZA (1946).

Sobre o comportamento mitótico e meiótico de cromossomas policêntricos ou com ponto de inserção difuso

Material: Studies were made mainly with Ascaris megalocephála Cloq. univalens and bivalens, and also with Tityus bahiensis Perty. 1) Somatic pairing of heterochromatic regions. The heterochromatic ends of the somatic chromosomes in Ascaris show a very strong tendency for unspecifical somatic pairing which may occur between parts of different chromosomes (Figs. 1, 2, 3, 7, 10, 11, 12, 13, 14, 16, 18,), between the two ends of the same chromosome either directly (Figs. 4, 5, 7, 8, 11, 12, 13, 15, 16, 17, 18) or inversely (Fig. 8, in the arrow) and also within a same chromosomal arm (Fig. 6). 2) During the early first cleavage division the chomosomes are an isodiametric cylinder (Figs. 6, 9, 11, 13, 14). But in later metaphase the ends become club shaped (Figs. 1, 2, 3, 4, 5, 7, 10) which is interpreted as the beginning of migration of chromatic substance from the central euchromatic region towards the heterochromatic regions. This migration becomes more and accentuated in anaphase (Figs. 19, 22, 23) and in the vegetative cells where euchromatic region looses more and more staing power, especially in the intersititial zones between the individual small spherical chromosomes into which the euchromatic region desintegrates. The emigrated chromatin material is finally eliminated with the heterochromatic chromosome ends (Fig. 23 and 24). 3) It seems a general rule that during mitotic anaphase all chromosomes with diffuse or multiple spindle fiber attachement (Ascaris, Tityus, Luzula, Steatococcus, Homoptera and Heteroptera in general) move to the poles in the form of an U with precedence of the chromosomal ends. In Ascaris, the heterocromatic regions are pulled passively towards the poles and only the euchromatic central portion may be U-shaped (Fig. 19, 22, 25). While in the other species this U-shape is perfect since the beginning of anaphase, giving the impression that movement towards the poles begins at both ends of a chromosome simultaneously, this is not the case in Ascaris. There the euchromatic region is at first U-shaped, passing then to form a straight or zig-zag line and becoming again U-shaped during late anaphase. This is explained by the fact that the ends of the euchromatic regions have to pull the weight of the passive heterochromatic portions. 4) While it is generally accepted that, during first meio-tic division untill second anaphase, all attachement regions remain either undivided or at least united closely, this is not the case in chromosomes with diffused or multiple attachment. Here one clearly sees in all cases so far studied four parallel chromatids at first metaphase. In Luzula and Tityus (for Tityus all figs. 26 to 31) this division is allready quite clear in paraphase (pro-metaphase) and it cannot be said wether in other species the division in sister chromatids is allready present, but not visible at this stage. During first anaphase the sister chromatids of Titbits remain more or less in contact, while in Luzula and especially in Ascaris they are quite separated. Thus one can count in late anaphase or telophase of Ascaris megalocephala bivalens, nearly allways, four separate chromosomes near each pole, or a total of eight chromatids per division figure (Figs. 35, 36, 37, 38, 39, 40, 41).

Provas cruciais da dicentricidade dos cromossômios dos hemipteros

Studying the meiosis of two Hemiptera, mamely, Lybindus dichrous (Coreidae) and Euryophthalmus humilis (Pyrrhocoridae), the author has found new proofs in favor of the existence of a centromere at each end of the chromosomes of the insects belonging to that order. Following the behaviour of a pair of large autosomes of Lybindus, he was able to verify that in the first division of the spermatocytes, the tetrad they form divides transversely by the middle, giving rise to two V-shaped anaphase chromosomes that go to the poles with the vertex pointing forwardly. From the end of the first division till the metaphase of the second one, the centromeres occupying the vertex of the V go apart from one another, making the chiasmata existing there slip to the opposite extremities, what changes the V into an X. When the chiasmata reach the acentric ends, the X is again converted into a V. The V of the secondary metaphase, therefore, differs from the V of the primary anaphase, in being inverted that is, in having the centromeres in the extremity of its arms, and no longer in the vertex as in the latter. The opening out of the chromosomes starting at the centric extremities in order to recuperate the dumbbell shape they show in the secondary anaphase, just in the manner postulated by PIZA, is thus demonstrated. In Euryophthalmus humilis it was verified once more, that the heterochromosome, in the secondary spermatocytes, orients parallelly to the spindle axis, accompanying with its ends the anaphase plates as they move to the poles. The author is in disagreement with NORONHA-WAGNER & DUARTE DE CASTRO's interpretation of the behaviour of the chromosomes in meiosis of Luzula nemorosa.

Nota sobre ácaros (Acari) atacando fruteira-do-conde (Rollinia sp.)

The flowers of this orchard tree presents six fleshy petals, fused two by two, and disposed propeller like. In the center is a cavity which harbors the cone shaped reproductive organs. An eriophyid mite, described by H. H. Keifer as Aculops flechtmann Keifer, 1972, causes considerable bronzing to the petals (Fig. 1-A). Large numbers of this mite attacking the basis of the reproductive organs causes them to dry and drop. On the young fruits, which are squamous, this mite causes some rusting and small necrotic areas (Fig. 1-B). As the fruit grows these small areas remain dark and barky. Very often the necrotic areas are invaded by the false spider mite Brevipalpus phoenicis (Geijskes, 1939), which enlarges the damage. The false spider mite also attacks leaves and green stems; the epidermis of the latter then assumes a barky appearance.

Nueva especie de Cephalobium (Rhabditida, Diplogasteridae) parasito de ninfas de Gryllodes Laplatae (Orthoptera, Gryllidae) en la Argentina

Cephalobium bidentatum sp. nov., parasitizing nymphs of Gryllodes laplatae Sauss (Orthoptera, Gryllidae) from Argentina, is described and illustrated. It is distinguished from other members of the genus Cephalobium COBB, 1920, by having the buccal cavity very sclerotized with two hook-shaped teeth, vagina short and muscular, male has two spicules with hook-shaped tips, and by the distribution pattern of the postanal papillae: one pair under the anus, three pairs between the anus and the tail, and two pairs at the base of the tail appendage.

Microspines in the pylorus of Pseudonannolene tricolor and Rhinocricus padbergi (Arthropoda, Diplopoda)

The aim of this paper is to report the morphology and distribution of microspines in diplopods pylorus, as these are important structures present along the alimentary tract of arthropods. The morphology of the internal surface of the pylorus of Pseudonannolene tricolor Brolemann, 1901 and Rhinocricus padbergi Verhoeff, 1938 was analyzed by SEM. Pseudonannolene tricolor presents two morphologically distinct pyloric regions: anterior and posterior. The first region is characterized by the presence of thin microspines that increase in number and size towards the posterior portion; the second region presents smaller and triangular-shaped microspines distributed throughout small plates. The pylorus of R. padbergi does not present differentiated regions; the anterior portion is characterized by microspines grouped in plates that decrease in number and increase in size towards the ileum.

Stridulatory file and calling song of two populations of the tropical bush cricket Eneoptera surinamensis (Orthoptera, Gryllidae, Eneopterinae)

This study characterizes the calling song and ultramorphology of the stridulatory file of two geographically isolated populations of the tropical bush cricket Eneoptera surinamensis (De Geer, 1773) from city of Foz do Iguaçu, state of Paraná, and town of Rio Claro, state of São Paulo, Brazil, distant 1,000 Km from each other. The teeth are shell-shaped, the larger ones are distributed in the medium region of the file, decreasing gradually in size towards the edges. Specimens from Foz do Iguaçu have a file with 82 ± 9.8 teeth, length=1.89 mm ± 0.15 with 43.76 ± 5.94 teeth per mm (n=15). Specimens from Rio Claro present a file with 87 ± 9.81, length=1.96 ± 0.19 mm with 44.52 ± 4.61 teeth per mm (n=15). Statistical differences found between the two populations are not significant. The calling song is an uninterrupted trill that alternates two sets of notes distinct for its temporal features.

Hexamermis paranaense new species (Nematoda, Mermithidae): a parasite of Diloboderus abderus (Coleoptera, Scarabaeidae) in Argentina

A new species, Hexamermis paranaense n. sp. (Nematoda, Mermithidae), a parasite of larvae of Diloboderus abderus Sturm, 1826 (Coleoptera, Scarabaeidae) is described and illustrated. This new species is characterized by amphids small, amphidial opening pocket-shaped, the anterior portion of the vagina muscularized and slightly protruding with a descending branch forming a loop before joining the uterus, and three rows of genital papillae: the ventrolateral divided in two rows with eight papillae in the outer row and with six papillae in the inner one; the ventral row with four pairs and one single preanal papillae, and with two pairs, a triplete, one pair, a single and one pair postanal papillae.

Reproductive behavior of Leptodactylus mystacinus (Anura, Leptodactylidae) with notes on courtship call of other Leptodactylus species

Here we present data on the reproductive behavior of Leptodactylus mystacinus (Burmeister, 1861), including details on courtship behavior. We also describe and compared the courtship calls of L. mystacinus, L. furnarius Sazima & Bokermann, 1978 and Leptodactylus sp. (L. aff. andreae). Field works were conducted in Uberlândia (central Brazil). During courtship, a female approaches a calling male and is led to a previously excavated chamber; a female can approach a silent male that beat his hands and/or feet on the ground as well. The courtship call of L. mystacinus consists of one single arch-shaped note (duration = 0.04 s) repeated 258 times per minute; the courtship calls of L. furnarius (0.06 s, 84 times per minute) and Leptodactylus sp. (0.15 s, 5 times per minute) also are arch-shaped. The courtship behavior of L. mystacinus is similar to that of other species of the L. fuscus (Schneider, 1799) group; unique to it is that males can beat his hands and/or feet on the ground while courting. The male behavior of conducting the female to a previously excavates chamber and the arch-shaped courtship call may represent other shared derived features of members of the L. fuscus group, including the former Adenomera species.

Two new Neotropical species of Drosophila peruensis species group (Diptera, Drosophilidae)

The Drosophila peruensis species group was recently proposed and includes four taxa: D. atalaia Vilela & Sene, 1982, D. boraceia Vilela & Val, 2004, D. pauliceia Ratcov & Vilela, 2007, and D. peruensis Wheeler, 1959. All these species have most of setae or setulae of mesonotum arinsing from dark spots, wings with crossveins darker (except in D. atalaia) and hypandrium squared-shaped mostly fused to gonopods. Here, we describe two new species, Drosophila itacorubi sp. nov. and Drosophila paraitacorubi sp. nov., belonging to this species group. The male genitalia of these species are figured. An identification key to the D. peruensis species group is provided.

On Munduruku, a new Theraphosid genus from Oriental Amazonia (Araneae, Mygalomorphae)

Munduruku gen. nov. is proposed for the type species Munduruku bicoloratum sp. nov., from Juruti and Santarém, Pará, Brazil. The main diagnostic character of Munduruku gen. nov. is the presence of a subapical, lanceolate keel on the male palpal bulb, which is unique among the basal taxa of Theraphosinae with type III-IV urticating setae. The female spermathecae consist of two spheroid receptacles with funnel-shaped necks, each of which bears a sclerotized area. In both sexes, the abdomen is remarkably patterned, an uncommon feature in adults of New World theraphosids. Both the bulbus lanceolate keel and the abdominal color pattern are hypothesized as synapomorphies of the genus.

Ensaios de Mycologia

Morphologically the fungus reminds one of the Microsporum species of man. The microscopic elements are: the shape of the mycelia within the hairs, size and disposition of spores intercalar and terminal chlamydospores, simple clusters, hyphae, pectinate and denticulate elements. Of the 17 species of Microsporum described up to now, none can be identified with Microsporum circuluscentrum. The microsporum described is of the pure-by human type. (Culture fairly vivacious, almost total absence of spindles.) Microscopically at first sight the culture might be taken for Microsporum fulvum URIBURU 1917 or for M. felineum. These are however from animals. The bean shaped culture on potatoes leaves a place apart for it among Microsporum fungi. The anomalous elements seen in the cultures are also found in other species of Microsporum. Besides the pseudo-perithecia described and interpreted I must mention that the pseudo-nodular organs are not an exception among these fungi, for Microsporum fulvum also shows what writers have described as nodular organs. The spiral elements are also not exclusive features of the trichophytes. Microsporum tardum also shows quite characteristic spiral forms.

Polarization in the chilean party system: changes and continuities, 1990-1999

The focus of this thesis is the evolution of programmatic polarization in the post-authoritarian Chilean party system at the elite level. It shows the distance/proximity between parties located along the left-right ideological continuum on three sets of issues. The paper demonstrates that important changes have taken place in the meaning of the right and, especially, left poles. This implies convergence on socio-economic issues between parties, but persistence of differences on religious-value issues, and on issues related to the authoritarian/democratic cleavage. Distance between the poles has been reduced, and as a result the center has lost its own political space. In addition, the paper shows that the pattern followed by programmatic polarization at the elite level is explained by the authoritarian experience, the institutional framework, and socio-economic transformations. Together with this factors, the degree of negotiability of the issues and the cross-cutting nature of the cleavages have also shaped polarization.