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The p-arms of the five human acrocentric chromosomes bear nucleolar organizer regions (NORs) comprising ribosomal gene (rDNA) repeats that are organized in a homogeneous tandem array and transcribed in a telomere-to-centromere direction. Precursor ribosomal RNA transcripts are processed and assembled into ribosomal subunits, the nucleolus being the physical manifestation of this process. I review current understanding of nucleolar chromosome biology and describe current exploration into a role for the NOR chromosomal context. Full DNA sequences for acrocentric p-arms are now emerging, aided by the current revolution in long-read sequencing and genome assembly. Acrocentric p-arms vary from 10.1 to 16.7 Mb, accounting for ∼2.2% of the genome. Bordering rDNA arrays, distal junctions, and proximal junctions are shared among the p-arms, with distal junctions showing evidence of functionality. The remaining p-arm sequences comprise multiple satellite DNA classes and segmental duplications that facilitate recombination between heterologous chromosomes, which is likely also involved in Robertsonian translocations. [ABSTRACT FROM AUTHOR]

In this study, we investigated the chromosomes of three species of Sicarius spiders from the Brazilian Caatinga, using classical and molecular cytogenetic techniques. Based on the phylogenetic approach, we also discussed about the variation of diploid number, types of sex chromosome system and changes in the localization of ribosomal genes of Scytodoidea. Sicarius are Synspermiata spiders that together with the genera Loxosceles and Hexophthalma constitute the family Sicariidae. In this group, the available cytogenetic data showed a low diploid number range (2n♂=18 to 2n♂=23) and the presence of only multiple sex chromosome systems (X1X2Y and X1X20). Mitotic metaphase cells exhibited 2n♂=16+X1X2Y for Sicarius cariri and S. ornatus, and 2n♂=18+XY for S. tropicus. In these species, silver impregnation revealed nucleolar organizer region (Ag-NOR) on the terminal region of pair 1. In S. ornatus and S. tropicus, the results obtained with fluorescent in situ hybridization (FISH) using 18S rDNA probe were similar to Ag-NOR, however in S. cariri, the ribosomal sites were localized in the terminal region of the X1 sex chromosome. In this work, we presented the first description of a simple sex chromosome system for Sicariidae, helping to understand how the XY sex chromosome system evolved from the X1X2Y system. Additionally, FISH data incongruous with Ag-NOR indicate that the cytogenetic studies in Sicariidae allow investigating the relation between the karyotype evolution and the distribution and the activity of rDNA genes.

Simple Summary: Dragonflies and damselflies (Odonata) are very interesting insects from a cytogenetic point of view. Their chromosomes do not have a typical centromere and their meiosis process differs in some respects from the canonical meiosis process. Sex in Odonata is usually determined by two X chromosomes in females and only one X chromosome in males (a Y chromosome is not present). In this work, we studied sex chromosome evolution in two dragonfly species of the genus Rhionaeschna that have a derived sex chromosome system: neo-XX in females and neo-XY in males. This variation is the result of chromosome rearrangements. In R. planaltica, meiotic analysis and fluorescence in situ hybridization with a ribosomal DNA probe revealed that the original X chromosome was inserted into the smallest autosome, giving rise to the neo-X chromosome, while the homologous autosome became a neo-Y chromosome. In contrast, the neo-X chromosome in R. bonariensis evolved by a terminal fusion of the original X chromosome with the largest autosome, whose homolog became the neo-Y chromosome. Our results suggest an independent origin of neo-sex chromosomes in these dragonfly species and contribute to our understanding of the distinct mechanisms of sex chromosome evolution. Odonata have holokinetic chromosomes. About 95% of species have an XX/X0 sex chromosome system, with heterogametic males. There are species with neo-XX/neo-XY sex chromosomes resulting from an X chromosome/autosome fusion. The genus Rhionaeschna includes 42 species found in the Americas. We analyzed the distribution of the nucleolar organizer region (NOR) using FISH with rDNA probes in Rhionaeschna bonariensis (n = 12 + neo-XY), R. planaltica (n = 7 + neo-XY), and Aeshna cyanea (n = 13 + X0). In R. bonariensis and A. cyanea, the NOR is located on a large pair of autosomes, which have a secondary constriction in the latter species. In R. planaltica, the NOR is located on the ancestral part of the neo-X chromosome. Meiotic analysis and FISH results in R. planaltica led to the conclusion that the neo-XY system arose by insertion of the ancestral X chromosome into an autosome. Genomic in situ hybridization, performed for the first time in Odonata, highlighted the entire neo-Y chromosome in meiosis of R. bonariensis, suggesting that it consists mainly of repetitive DNA. This feature and the terminal chiasma localization suggest an ancient origin of the neo-XY system. Our study provides new information on the origin and evolution of neo-sex chromosomes in Odonata, including new types of chromosomal rearrangements, NOR transposition, and heterochromatin accumulation. [ABSTRACT FROM AUTHOR]

Odonata have holokinetic chromosomes. About 95% of species have an XX/X0 sex chromosome system, with heterogametic males. There are species with neo-XX/neo-XY sex chromosomes resulting from an X chromosome/autosome fusion. The genus Rhionaeschna includes 42 species found in the Americas. We analyzed the distribution of the nucleolar organizer region (NOR) using FISH with rDNA probes in Rhionaeschna bonariensis (n = 12 + neo-XY), R. planaltica (n = 7 + neo-XY), and Aeshna cyanea (n = 13 + X0). In R. bonariensis and A. cyanea, the NOR is located on a large pair of autosomes, which have a secondary constriction in the latter species. In R. planaltica, the NOR is located on the ancestral part of the neo-X chromosome. Meiotic analysis and FISH results in R. planaltica led to the conclusion that the neo-XY system arose by insertion of the ancestral X chromosome into an autosome. Genomic in situ hybridization, performed for the first time in Odonata, highlighted the entire neo-Y chromosome in meiosis of R. bonariensis, suggesting that it consists mainly of repetitive DNA. This feature and the terminal chiasma localization suggest an ancient origin of the neo-XY system. Our study provides new information on the origin and evolution of neo-sex chromosomes in Odonata, including new types of chromosomal rearrangements, NOR transposition, and heterochromatin accumulation.

Background Oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL) with dysplasia are closely linked conditions in the oral cavity, with the latter often indicating precancerous changes, underscoring the urgency of early detection and intervention. Histopathological confirmation is crucial for accurate diagnosis. The nucleolar organizer region (NOR), specifically analyzed through silver-staining (argyrophilic NORs), provides insights into nuclear changes associated with the lesion. Computer-assisted morphometric analysis enhances precision and objectivity in evaluating AgNOR-related parameters. Aim To conduct a computer-assisted morphometric comparison of AgNORs using various NOR-related parameters in cases of OSCC and leukoplakia with dysplasia and to evaluate their diagnostic significance. Materials and methods A computer-assisted morphometric analysis was conducted using various NOR-related parameters, such as nuclear profile area, single AgNOR profile area per nucleus, total AgNOR profile area per nucleus, and number of AgNOR profiles per nucleus on a total sample of 90 specimens, which includes leukoplakia with dysplasia (30), OSCC (30), and a control group, including 30 samples of normal oral mucosa. A comparison was conducted on the morphometric values between the groups under investigation. Tukey's multiple comparison tests and ANOVA were used to analyze the data and determine the differences between the groups. Results The present investigation revealed a significant difference in all four AgNOR-related parameters between leukoplakia and OSCC in comparison to the control group (normal oral mucosa). Comparing OL (41.78 ± 0.46) and OSCC (62.78 ± 0.47) to the control group (35.93 ± 0.99), the mean value of nuclear profile area (A Nuc) was significantly greater. In comparison to the control group (3.40 ± 0.09), the mean value of a single AgNOR profile area per nucleus (A NOR) was found to be relatively lower in both research groups, OL (2.00 ± 0.02) and OSCC (1.39 ± 0.01). The total AgNOR profile area per nucleus (TA NOR) had a mean value of 10.61 ± 0.69 in OL and 12.05 ± 0.28 in OSCC, respectively, compared to 7.82 ± 0.38 in the control group. The study found that there was more number of profiles of AgNORs per nucleus (n NOR) in the study groups of OL (5.30 ± 0.29) and OSCC (8.69 ± 0.19) than in the control group (2.32 ± 0.11). Conclusion The parameters linked to the NOR are biologically informative and easy to check regularly in a pathology lab. Additionally, AgNORs give us important information that enables us to study the range of nuclear changes in malignant and potentially malignant lesions., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Nararyanan et al.)

Whip spiders (Amblypygi) represent an ancient order of tetrapulmonate arachnids with a low diversity. Their cytogenetic data are confined to only a few reports. Here, we analyzed the family Charinidae, a lineage almost at the base of the amblypygids, providing an insight into the ancestral traits and basic trajectories of amblypygid karyotype evolution. We performed Giemsa staining, selected banding techniques, and detected 18S ribosomal DNA and telomeric repeats by fluorescence in situ hybridization in four Charinus and five Sarax species. Both genera exhibit a wide range of diploid chromosome numbers (2n = 42–76 and 22–74 for Charinus and Sarax, respectively). The 2n reduction was accompanied by an increase of proportion of biarmed elements. We further revealed a single NOR site (probably an ancestral condition for charinids), the presence of a (TTAGG)n telomeric motif localized mostly at the chromosome ends, and an absence of heteromorphic sex chromosomes. Our data collectively suggest a high pace of karyotype repatterning in amblypygids, with probably a high ancestral 2n and its subsequent gradual reduction by fusions, and the action of pericentric inversions, similarly to what has been proposed for neoamblypygids. The possible contribution of fissions to charinid karyotype repatterning, however, cannot be fully ruled out.

In this study, we examined for the first time the distribution of the 28S ribosomal genes in beetles of the subfamily Cassidinae. More than 55% of the species in this subfamily have a similar karyotype, 2n = 16 + Xyp. For this work, we selected species belonging to the tribes Cassidini and Mesomphaliini, which have, respectively, the most conserved and diversified karyotype characteristics within the Cassidinae. An analysis of 11 species revealed that rDNA sites on one pair of autosomes is the most frequent pattern, occurring in 10 species. This condition occurs in the seven genera examined and in species of both of the tribes, Cassidini and Mesomphaliini. Nevertheless, the differences in the locations of 28S rDNA were more pronounced in the tribe Cassidini and among species with similar karyotype characteristics. On the other hand, in Mesomphaliini, the increase in the diploid number was not accompanied by an increase in the number of ribosomal sites. Moreover, the comparison of the number and localization of major rDNA sites with the distribution of constitutive heterochromatin indicates that there is no direct correlation between the dispersion of constitutive heterochromatin and 28S rDNA genes in Cassidinae.

A wide range of diploid number of chromosomes and the body size of Channa congeners are useful combination of characters for studying the factors controlling the body size. In this study, the karyological information was superimposed on the evolutionary tree generated by 16S rRNA mitochondrial gene sequences. Here, the metaphase chromosome complements stained with Giemsa, AgNO3 and CMA3 were prepared from six snakehead murrel fish species collected from northeast India. The diploid chromosome numbers and the fundamental arms of C. aurantimaculata (2n = 52, NF = 98), C. gachua (2n = 56, NF = 84), C. marulius (2n = 44, NF = 58), C. orientalis (2n = 52, NF = 74), C. punctata (2n = 32, NF = 60) and C. striata (2n = 40, NF = 48) were calculated by the analysis of metaphase chromosome complements. Both methods of nucleolar organizer region (NOR) localization, silver nitrate and chromomycin A3, revealed NOR pairs of 1, 2, 3, 1, 4 and 3 in C. aurantimaculata, C. gachua, C. marulius, C. orientalis, C. punctata and C. striata, respectively. The subject species showed primitive type of asymmetrical chromosomes, except the C. punctata. The variation in 2n for C. orientalis (2n = 52, 78) and C. gachua (2n = 52, 78, 104) of a complete haploid set indicates the possibility of either ploidy change in C. orientalis and C. gachua, if we consider 2n = 52 or the Robertsonian rearrangements in different populations of these two species. The chromosome evolution tree was constructed on 16S rRNA ML-phylogenetic tree using ChromEvol 1.3. The analysis of chromosome evolution explained the loss or gain of chromosome, duplications or semiduplications mechanism. For time scaling the chromosome evolution, the node age of available 16S rRNA gene of Channa species were estimated, which was also used for estimating the time when chromosomal changes occurred in context of geological time-scale. [ABSTRACT FROM AUTHOR]

Changing of structural and quantitative parameters of fibrillar centers and nucleolar organizer regions in nucleolus are a biological marker of cancer. These mentioned areas are identified by using specific argyrophilic proteins (RNA-polymerase I, UBF, Nucleolin C23, nucleophosmin I or B23), silver staining (AgNOR) and immunocytochemistry methods. The argyrophilic proteins of the nucleolus are actively used in scientific and clinical practice to assess the different functional activity of the cell, especially cell cycle, transcription and malignant transformation processes. Increasing the number and size of nucleolar organizer regions confirms the increased biosynthetic activity of the nucleolus in the normal cell or the activation of the malignant transformation processes during the growth of the cancer. Increasing argyrophilic proteins expression caused by morphological changes presents the marker for determination of malignant cancer cells and proliferative activity. [ABSTRACT FROM AUTHOR]

Summary : The reference genome sequence of wheat 'Chinese Spring' (CS) is now available (IWGSC RefSeq v1.0), but the core sequences defining the nucleolar organizer regions (NORs) have not been characterized. We estimated that the total copy number of the rDNA units in the wheat genome is 11 160, of which 30.5%, 60.9% and 8.6% are located on Nor‐B1 (1B), Nor‐B2 (6B) and other NORs, respectively. The total length of the NORs is estimated to be 100 Mb, corresponding to approximately 10% of the unassembled portion of the genome not represented in RefSeq v1.0. Four subtypes (S1–S4) of the rDNA units were identified based on differences within the 3′ external transcribed spacer regions in Nor‐B1 and Nor‐B2, and quantitative PCR indicated locus‐specific variation in rDNA subtype contents. Expression analyses of rDNA subtypes revealed that S1 was predominantly expressed and S2 weakly expressed, in contrast to the relative abundance of rDNA subtypes in the wheat genome. These results suggest a regulation mechanism of differential rDNA expression based on sequence differences. S3 expression increased in the ditelosomic lines Dt1BL and Dt6BL, suggesting that S3 is subjected to chromosome‐mediated silencing. Structural differences were detected in the regions surrounding the NOR among homoeologous chromosomes of groups 1 and 6. The adjacent regions distal to the major NORs were expanded compared with their homoeologous counterparts, and the gene density of these expanded regions was relatively low. We provide evidence that these regions are likely to be important for autoregulation of the associated major NORs as well as silencing of minor NORs. Significance Statement: Based on the wheat reference genome sequence (IWGSC RefSeq v1.0), we characterized the structure of two major nucleolus organizer regions, loci encoding rRNAs, and their surrounding regions in the wheat genome and identified the composition of rDNA units that display chromosome‐specific expression patterns. These results lead to an understanding of the nucleolar dominance associated with allopolyploidization during the evolution of wheat. [ABSTRACT FROM AUTHOR]

Background Silver stainable nucleolar organizer regions (AgNORs) have proven to exhibit utmost importance due to their higher occurrence in the nucleus especially in malignant cells than in normal. Thus, they assist in the examination of nucleolar structures and variations in nucleolar activity. Aim Quantitative and qualitative analysis in relation to the number and area of AgNOR in tissue sections of the normal oral mucosa (NOM), oral leukoplakia (OL), and oral squamous cell carcinoma (OSCC) was the main aim of the study. Materials & method A total of 50 cases comprising 20 OL with dysplasia, 20 OSCC cases, and 10 samples of normal oral mucosa were taken. Silver nitrate (Sol A) & gelatin (Sol B) solutions were freshly prepared for staining the lesional slides. Results The mean value of nuclear profile area (A Nuc) was comparatively higher in oral leukoplakia i.e. 41.97 and in oral squamous cell carcinoma i.e. 62.36 in comparison to the control group where it was 36.19. The mean value of a single AgNOR profile area per nucleus (A NOR) was found to be comparatively lower in both study groups i.e. oral leukoplakia (2.76) and oral squamous cell carcinoma (1.61) in comparison to the control group (3.45) . The mean value of total AgNOR profile area per nucleus (TA NOR) and the number of profiles of AgNORs per nucleus (n NOR) were found higher in both study groups (oral leukoplakia and oral squamous cell carcinoma) as compared to normal oral mucosa of the control group. However, the findings of all four parameters of morphometric analysis were found to be significantly associated with disorder of oral mucosa i.e. cases of oral leukoplakia and oral squamous cell carcinoma (P value <0.01). Conclusion It can thus be suggested that the mean AgNOR count displayed a higher value in OSCC. Hence, the number of AgNORs in nuclei increases as epithelial cells undergo malignant transformation which is designated that mean AgNOR count may contribute to establishing the prognosis of a lesion., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Singh et al.)

In this study, we examined for the first time the distribution of the 28S ribosomal genes in beetles of the subfamily Cassidinae. More than 55% of the species in this subfamily have a similar karyotype, 2n = 16 + Xyp. For this work, we selected species belonging to the tribes Cassidini and Mesomphaliini, which have, respectively, the most conserved and diversified karyotype characteristics within the Cassidinae. An analysis of 11 species revealed that rDNA sites on one pair of autosomes is the most frequent pattern, occurring in 10 species. This condition occurs in the seven genera examined and in species of both of the tribes, Cassidini and Mesomphaliini. Nevertheless, the differences in the locations of 28S rDNA were more pronounced in the tribe Cassidini and among species with similar karyotype characteristics. On the other hand, in Mesomphaliini, the increase in the diploid number was not accompanied by an increase in the number of ribosomal sites. Moreover, the comparison of the number and localization of major rDNA sites with the distribution of constitutive heterochromatin indicates that there is no direct correlation between the dispersion of constitutive heterochromatin and 28S rDNA genes in Cassidinae. [ABSTRACT FROM AUTHOR]

Nucleoli are the sites of ribosome biogenesis and the largest membraneless subnuclear structures. They are intimately linked with growth and proliferation control and function as sensors of cellular stress. Nucleoli form around arrays of ribosomal gene ( rDNA) repeats also called nucleolar organizer regions ( NORs). In humans, NORs are located on the short arms of all five human acrocentric chromosomes. Multiple NORs contribute to the formation of large heterochromatin-surrounded nucleoli observed in most human cells. Here we will review recent findings about their genomic architecture. The dynamic nature of nucleoli began to be appreciated with the advent of photodynamic experiments using fluorescent protein fusions. We review more recent data on nucleoli in Xenopus germinal vesicles ( GVs) which has revealed a liquid droplet-like behavior that facilitates nucleolar fusion. Further analysis in both Xenopus GVs and Drosophila embryos indicates that the internal organization of nucleoli is generated by a combination of liquid-liquid phase separation and active processes involving rDNA. We will attempt to integrate these recent findings with the genomic architecture of human NORs to advance our understanding of how nucleoli form and respond to stress in human cells. [ABSTRACT FROM AUTHOR]

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize karyotypes of central European spiders of the genera Arctosa, Tricca, and Xerolycosa (Lycosidae) with respect to the diploid chromosome number, chromosome morphology, and sex chromosomes. Karyotype data are reported for eleven species, six of them for the first time. For selected species the pattern in the distributions of the constitutive heterochromatin and the nucleolar organizer regions (NORs) was determined. The silver staining technique for detecting NORs of lycosid spiders was standardized. The male karyotype consisted of 2n = 28 (Arctosa and Tricca) or 2n = 22 (Xerolycosa) acrocentric chromosomes. The sex chromosome system was X1X20 in all species. The sex chromosomes of T. lutetiana and X. nemoralis showed unusual behaviour during late diplotene, namely temporary extension due to decondensation. C-banding technique revealed a small amount of constitutive heterochromatin at the centromeric region of the chromosomes. Two pairs of autosomes bore terminal NORs. Differences in karyotypes among Arctosa species indicate that the evolution of the karyotype in this genus involved autosome translocations and size changes in the sex chromosomes. Based on published results and those recorded in this study it is suggested that the ancestral male karyotype of the superfamily Lycosoidea consisted of 28 acrocentric chromosomes.

Theridiidae is a derived family within the Araneoidea clade. In contrast to closely related groups, the 2n(male) = 20+X1X2 with acro/telocentric chromosomes is the most widespread karyotype among the theridiid spiders. In this work, the cytogenetic analysis of Argyrodes elevatus revealed original chromosome features different from those previously registered for Theridiidae, including the presence of 2n(male) = 20+X with meta/submetacentric chromosomes. Most individuals of Nesticodes rufipes showed family conserved karyotype characteristics. However, one individual had a 2n(male) = 24 due to the presence of an extra chromosome pair, which exhibited regular behavior and reductional segregation during meiosis. After silver staining, mitotic cells exhibited NORs localized on the terminal regions of the short arms of pairs 2, 3, and 4 of A. elevatus and on the terminal regions of long arms of pair 4 of N. rufipes. The comparative analysis with data from phylogenetically related species allowed the clarification of the origin of the interspecific and intraspecific chromosome variability observed in Argyrodes and in N. rufipes, respectively.

Nephilid systematics has been subject to several changes in the last years, and the use of non-classical characters could be useful for evolutionary considerations. In this study, we analyzed the mitotic chromosomes of two nephilid spiders,Nephila clavipesandNephila sexpunctata, using standard staining, silver nitrate impregnation and C-banding techniques, aiming to discuss the chromosomal similarities of Nephilidae and Tetragnathidae, and chromosome evolution withinNephilaandNephilingis. The basic karyotype characteristics observed in these two species (2n♂ = 22 + X1X20 and monoarmed chromosomes) were similar to those registered for most araneoid families, i.e., Araneidae, Linyphiidae, Nephilidae, Nesticidae and Tetragnathidae. However, the occurrence of both prominent secondary constrictions and nucleolar organizer regions (NORs) is a shared characteristic between Nephilidae and Tetragnathidae, considering that these regions were not observed in any other Araneoidea species cytogenetically examined. Furthermore, in the present study we showed that withinNephilaandNephilingisspecies, change in the number and location of NORs as well as in the quantity and distribution of constitutive heterochromatin were the main events responsible for chromosome evolution, and that these differences can be useful in the cytotaxonomy of this group. [ABSTRACT FROM AUTHOR]

Fluorescence in situ hybridization of Anopheles darlingi and A. nuneztovari demonstrated nucleolar organizer region activity at the end of the fourth larval instar, when the nucleolar organizer regions underwent gradual condensation. The heteromorphic sex chromosomes showed intraindividual size variation in the rDNA blocks located in the pericentromeric region and this coincided with the location of constitutive heterochromatin (C-banding).