Chromosomal characteristics of a Brazilian whip spider (Amblypygi) and evolutionary relationships with other arachnid orders.

We analyzed mitotic and meiotic cells of a Brazilian amblypygid, Heterophrynus longicornis, using conventional and molecular cytogenetic techniques (Giemsa staining, C-banding, Ag-NOR, and FISH with rDNA probe). This is the first study that focuses solely on amblypygid chromosomes; it was undertaken to add data on cytogenetic knowledge of this group and contribute to the understanding of chromosome evolution in the Arachnida. We found 2n = 66 for male and female individuals, monocentric chromosomes, and absence of morphologically differentiated sex chromosomes. C-banding showed heterochromatin in the pericentromeric region of most chromosomes. Mitotic and meiotic nuclei submitted to silver impregnation and FISH revealed, respectively, Ag-NORs and ribosomal genes in the terminal region of two chromosome pairs. Most chromosome features that we observed in H. longicornis are shared with species of other arachnid orders; however, the absence of morphologically differentiated sex chromosomes in amblypygid contrasts with the remarkable variety of sex chromosome systems recorded for the Araneae. Consequently, we conclude that analysis of species of the Tetrapulmonata clade is useful for understanding the trends of sex chromosome evolution in this arachnid group.

Chromosomal characteristics and karyotype evolution of Oxyopidae spiders (Araneae, Entelegynae).

We made a cytogenetic analysis of four species of Oxyopidae and compared it with the karyotype data of all species of this family. In Hamataliwa sp, the mitotic cells showed 2n♂ = 26+X(1)X(2) and telocentric chromosomes. The 2n♂ = 28, which has been described for only one oxyopid spider, is the highest diploid number reported for this family. Peucetia species exhibited distinct karyotype characteristics, i.e., 2n♂ = 20+X(1)X(2) in P. flava and 2n♂ = 20+X in P. rubrolineata, revealing interspecific chromosome variability within this genus. However, both Peucetia species exhibited telocentric chromosomes. The most unexpected karyotype was encountered in Oxyopes salticus, which presented 2n♂ = 10+X in most individuals and a predominance of biarmed chromosomes. Additionally, one male of the sample of O. salticus was heterozygous for a centric fusion that originated the first chromosomal pair and exhibited one supernumerary chromosome in some cells. Testicular nuclei of Hamataliwa sp and O. salticus revealed NORs on autosomal pairs, after silver impregnation. The majority of Oxyopidae spiders have their karyotype differentiated by both reduction in diploid number chromosome number and change of the sex chromosome system to X type; however, certain species retain the ancestral chromosome constitution 2n = 26+X1X2. The most remarkable karyotype differentiation occurred in O. salticus studied here, which showed the lowest diploid number ever observed in Oxyopidae and the second lowest registered for Entelegynae spiders.

Chromosomal characteristics and karyotype evolution of Oxyopidae spiders (Araneae, Entelegynae)

number chromosome number and change of the sex chromosomesystem to X type; however, certain species retain the ancestralchromosome constitution 2n = 26+X1X2. The most remarkablekaryotype differentiation occurred in O. salticus studied here, whichshowed the lowest diploid number ever observed in Oxyopidae andthe second lowest registered for Entelegynae spiders.

Intriguing evidence of translocations in Discus fish (Symphysodon, Cichlidae) and a report of the largest meiotic chromosomal chain observed in vertebrates.

As part of a program to understand the genetics of Amazonian ornamental fish, classical cytogenetics was used to analyze Symphysodon aequifasciatus, S. discus and S. haraldi, popular and expensive aquarium fishes that are endemic to the Amazon basin. Mitotic analyses in Symphysodon have shown some odd patterns compared with other Neotropical cichlids. We have confirmed that Symphysodon species are characterized by chromosomal diversity and meiotic complexity despite the fact that species share the same diploid number 2n=60. An intriguing meiotic chromosomal chain, with up to 20 elements during diplotene/diakinesis, was observed in S. aequifasciatus and S. haraldi, whereas S. discus only contains typical bivalent chromosomes. Such chromosomal chains with a high number of elements have not been observed in any other vertebrates. We showed that the meiotic chromosomal chain was not sex related. This observation is unusual and we propose that the origin of meiotic multiples in males and females is based on a series of translocations that involved heterochromatic regions after hybridization of ancestor wild Discus species.

Mesabolivar brasiliensis (Moenkhaus 1898) and Mesabolivar cyaneotaeniatus (Keyserling 1891) (Araneomorphae, Pholcidae) Close relationship reinforced by cytogenetic analyses

Pholcidae is the most diverse family among haplogyne spiders but only 15 species have been analyzed cytogenetically. These studies revealed that the diploid number varies from 2n = 15 to 2n = 32, that there are three types of sex chromosome systems in males (X, X1X2 and X 1X2Y), and that the chromosomes are predominantly biarmed. Within the genus Mesabolivar, only Mesabolivar luteus (Keyserling 1891) has been karyotyped, and it showed 2n = 15 = 14 + X, with all chromosomes being metacentric. In the present work, we characterize the mitotic and meiotic chromosomes of Mesabolivar brasiliensis (Moenkhaus 1898) and Mesabolivar cyaneotaeniatus (Keyserling 1891). Male mitotic metaphases of the two species showed the diploid number 2n = 17= 16 + X; oogonial cells of M. brasiliensis showed 2n = 18= 16 + XX. In both species, the chromosomes were exclusively biarmed, and the X chromosome was the largest element of the karyotype. Diplotene spermatocytes of the two species exhibited 8II + X and the occurrence of only one terminal or interstitial chiasma per bivalent. In M. cyaneotaeniaus, metaphases II with n = 9 = 8 + X and n = 8 were found, indicating the regular segregation of all chromosomes during meiosis I. Mitotic metaphases of M. brasiliensis stained with CMA3/DA/DAPI revealed GC-rich chromatin in the terminal region of almost all autosomes, especially in pair 2. An earlier revision of the New World pholcids grouped M. brasiliensis and M. cyaneotaeniatus in a "southern group" and placed M. luteus in a "miscellaneous group." A molecular study showed a closer relationship between M. brasiliensis and M. cyaneotaeniatus than between M. luteus and either of these two species. The 2n = 17 found in M. brasiliensis and M. cyaneotaeniatus corroborates this hypothesis, given that M. luteus has a diploid number of 2n = 15.

Cytogenetic analysis of the neotropical spider Nephilengys cruentata (Araneomorphae, tetragnathidae): standard staining, NORs, C-bands and base-specific fluorochromes

O objetivo deste trabalho é caracterizar Nephilengys cruentata em relação ao número diplóide, à morfologia cromossômica, ao tipo de sistema cromossômico de determinação sexual, aos cromossomos portadores de Regiões Organizadoras de Nucléolo (RONs), padrão de bandas C e seqüências AT ou GC repetitivas. As preparações cromossômicas foram submetidas à coloração convencional (Giemsa), à impregnação pelo nitrato de prata, técnica de obtenção de bandas C e à coloração com fluorocromos base-específicos. A análise das células mostrou 2n = 24 e 2n = 26 cromossomos nos embriões e 2n = 26 nas células ovarianas, sendo todos cromossomos acrocêntricos. O braço longo dos pares 1, 2 e 3 apresentou extensa região heteropicnótica negativa quando as metáfases mitóticas foram coradas com Giemsa. Os cromossomos sexuais não mostraram características diferenciais que permitissem distingui-los dos outros cromossomos do complemento. Considerando os números diplóides encontrados em N. cruentata e a predominância do sistema cromossômico de determinação sexual do tipo X1X2 em Tetragnathidae, N. cruentata parece contar com 2n = 24 = 22 + X1X2 nos machos e com 2n = 26 = 22 + X1X1X2X2 nas fêmeas. Os pares 1, 2 e 3 mostraram RONs coincidentes com as regiões heteropicnóticas negativas. Utilizando a técnica de obtenção de bandas C, a região pericentromérica dos cromossomos revelou pequena quantidade ou até mesmo ausência de heterocromatina constitutiva, diferindo do padrão de bandas C descrito em outras espécies de aranhas. Em N. cruentata, os fluorocromos DAPI/DA, DAPI/MM e CMA3/DA revelaram que a heterocromatina constitutiva é rica em bases AT e as RONs apresentam seqüências repetidas de bases GC.

Chromosomal data of two pholcids (Araneae, Haplogynae) A new diploid number and the first cytogenetical record for the New World clade

Mesabolivar luteus (Keyserling 1891) and Micropholcus fauroti (Simon 1887) specimens were collected in Ubatuba and Rio Claro, both in the state of São Paulo, Brazil. Mesabolivar luteus showed 2n (♂) = 15 = 14 + X and 2n (♀) = 16 = 14 + XX in mitotic metaphases and 7II + X in diplotenic cells. During late prophase I, all bivalents presented a ring shape, evidencing two chiasmata per bivalent. In this species, some diplotenic cells appear in pairs, maybe due to specific characteristics of the intercellular bridges. The metaphases II showed n = 7 or n = 8 = 7 + X chromosomes. Micropholcus fauroti evidenced 2n (♂) = 17 = 16 + X in spermatogonial metaphases and 8II+X in diplotenic cells, with only one chiasma per bivalent, contrasting with M. luteus. In both species, all chromosomes were metacentrics. The sexual chromosome X was the largest element and appeared as a univalent during meiosis I. These are the first cytogenetical data for the genera Mesabolivar and Micropholcus. Additionally, M. luteus is the first chromosomally analyzed species of the New World clade and the observed diploid number for M. fauroti had not yet been recorded in Pholcidae.

Recombination nodules in coleopteran species: Palembus dermestoides (Tenebrionidae) and Epicauta atomaria (Meloidae).

This work describes the first report about the occurrence of recombination nodules (RNs) in spread pachytene cells of two species of Coleoptera: Palembus dermestoides (Tenebrionidae) and Epicauta atomaria (Meloidae). The RNs were observed in preparations contrasted with phosphotungstic acid. Considering RN morphology and its occurrence in pachytene bivalents (one per autosome bivalent) these structures were interpreted to be late RNs. P. dermestoides and E. atomaria have 2n = 20 chromosomes including an Xy(p) sex determination system. In spite of most frequently subtelocentric morphology observed in the autosomes of both species, the occurrence of RNs is limited only to the synaptonemal complex (SC) structure of the long arms. These findings are in agreement with those obtained using light microscopy analysis in which only one chiasma or terminalization event is observed per autosomal bivalent in early or late metaphase I cells. The RNs have the same average width of the SC of each analyzed species, a circular shape, strong electron density, and are observed mainly between the lateral elements of the SC. The RNs of P. dermestoides and E. atomaria have approximately the same average size (width), 180 +/- 20 nm and 160 +/- 80 nm, respectively. The absence of RNs in the short arms and its occurrence in the long arms are discussed considering the short arm pericentromeric and pro-centric heterochromatin.

Cytogenetic analysis of Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) with Xyp sex determination system using standard staining, C-bands, NOR and synaptonemal complex microspreading techniques.

The mitotic and meiotic chromosomes of the beetles Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) were analysed using standard staining, C-banding and silver impregnation techniques. We determine the diploid and haploid chromosome numbers, the sex determination system and describe the chromosomal morphology, the C-banding pattern and the chromosome(s) bearing NORs (nucleolar organizer regions). Both species shown 2n = 20 chromosomes, the chromosomal meioformula 9 + Xyp, and regular chromosome segregation during anaphases I and II. The chromosomes of E. atomaria are basically metacentric or submetacentric and P. dermestoides chromosomes are submetacentric or subtelocentric. In both beetles the constitutive heterochromatin is located in the pericentromeric region in all autosomes and in the Xp chromosome; additional C-bands were observed in telomeric region of the short arm in some autosomes in P. dermestoides. The yp chromosome did not show typical C-bands in these species. As for the synaptonemal complex, the nucleolar material is associated to the 7th bivalent in E. atomaria and 3rd and 7th bivalents in P. dermestoides. Strong silver impregnated material was observed in association with Xyp in light and electron microscopy preparations in these species and this material was interpreted to be related to nucleolar material.