Studies on the chromosomes and sex chromatin in the horse

This study provides accumulated data to assist the definition of karyotypes from normal and infertile horses. The normal karyotype of the horse (2n = 64) was characterized following Giemsa staining and C- banding, and 23% aneuploidy was found among chromosome counts of cells prepared from 44 clinically normal horses and 24 equine embryos. These expected variations in chromosome counts are especially important in the evaluation of potential mosaicism. Centromere staining was shown to be a valuable aid for the identification of specific chromosomes, in particular the sex chromosomes. Sex chromatin studies were applied to nerve tissue and polymorphonuclear neutrophils obtained from three horses. Distinctive sex chromatin bodies were detected in 70% of neurones from a normal mare. The sex chromatin was most frequently located adjacent to the nucleolus. Nuclear appendages ("drumsticks") were present in 4% of polymorphonuclear neutrophils from a normal mare. Small numbers of similar structures were noted in the neutrophils from each animal examined.     

Seasonal sex ratio and unbalanced investment sex ratio in the Banksia bee Hylaeus alcyoneus

Abstract 1. Hylaeus alcyoneus is an endemic solitary bee common on coastal heaths of Western Australia. The bee is unusual in that males are larger than females. This size dimorphism presents an opportunity to test the theory of resource-dependent sex allocation, in which theory predicts that when resources are low the sex ratio should be biased towards the smaller sex. In most bees, females are larger than males and, in line with theoretical prediction, sex ratios are male biased when resources are scarce.
2. The emerging sex ratio and brood mass from a natural population of H. alcyoneus using trap nests was studied over two seasons (1999, 2000). A switch from a male- to a female-biased sex ratio through the season was found, which was related to a reduced floral resource.
3. Fisherian sex ratio theory predicts that total investment in each sex throughout a season should be equal and that the sex ratio should be biased towards the smaller sex. By measuring the mass of the emerging progeny, the total investment was found to favour males. Possible explanations for this bias in investment are discussed.     

A triple-primer PCR approach for the sex identification of endangered Phasianidae birds

To manage and conserve endangered Phasianidae species and to study their ecology, behavior, and population structure, sex identification is necessary. A search for the pheasant sex-linked marker (PSL) was carried out by comparing the sequences of chicken sex chromosomes. A triple-primer PCR approach was established in the present study based on a novel PSL for the sex identification of Phasianidae birds. The triple-primer system amplified a 202-bp PSL-W-specific fragment in female Phasianidae birds, whereas there is a 107-bp PSL-Z-specific fragment in the both sexes, thus making the female distinct. We also found a 102-bp AT-rich repeat sequence inserted into the PSL-W-specific region in Tetraophasis szechenyii generating a ∼300-bp PSL-W fragment in females. The sex of 11 Phasianidae species was distinguished clearly by the triple-primer PCR approach indicating that this method is both quick and reliable and can be easily applied to a wider range of Phasianidae species. Due to the relatively small size of the amplification products, the triple-primer PCR approach can also be applied to noninvasive samples such as feathers.     

Morphological and genetic sex identification of white-tailed eagle Haliaeetus albicilla nestlings

Identifying the sex of bird nestlings is relevant to studies of behaviour and ecology and is often a central issue in the management of endangered or captive populations. The white-tailed eagle Haliaeetus albicilla is a formerly threatened Eurasian raptor which is closely monitored in many countries due to its high exposure to environmental pollutants in the food chain. The aim of this study was to evaluate the reliability of sex identification methods for white-tailed eagle nestlings based on morphological measurements that can be recorded at the nest by a single person and with minimum disturbance. The sex of each bird was independently determined using molecular (genetic) methods. One measure of tarsus width allowed the correct identification of sex for 96% of the nestlings from southern and central Sweden. However, we found that the criteria for sex identification were not directly applicable to the population in Swedish Lapland, where nestlings are typically thinner, probably due to a limited food supply. These results show that sexing in the field of white-tailed eagle nestlings can be feasible with high accuracy based on a limited number of measurements. However, the criteria employed to separate sexes may have to be adjusted for each population.     

AFLP-derived STS markers for the identification of sex in Asparagus officinalis L.

For a simple, rapid and PCR-based screening of sex in the cultivated asparagus (Asparagus officinalis L.), we developed five STS markers from previously mapped, low-copy, sex-linked AFLP markers. A male/female PCR assay was feasible with these STS markers either by direct amplification or by digestion with restriction enzymes. Similar to the AFLP markers from which they were derived, STS4150.1, STS4150.2, STS4150.3 and STS3156 did not give recombinants in five different populations. STS3660 could be scored codominantly, enabling the differentiation of XY from YY males in the screened F₂ mapping population. The use of the sex-linked STS markers should allow early identification of sex, thus accelerating the breeding process for new asparagus varieties. Further, 10 additional AFLP markers obtained with PstI/MseI primer combinations have been mapped on the L5 chromosome, bringing the total number of known AFLP and STS markers flanking the sex locus to 24. These markers can be utilized for fine mapping of the sex gene in asparagus, which will pave the way for a map-based cloning approach. Received: 31 May 1999 / Accepted: 22 June 1999     

No evidence of mutations in four candidate genes for male sex determination/differentiation in sex-reversed XY females with campomelic dysplasia.

Campomelic dysplasia (Cd) occurs combined with sex reversal resulting in XY females. The recent identification of candidate genes for sex determination/differentiation and of a sex determining region on the human Y chromosome prompted the authors to study these genes for mutations in patients with Cd and sex reversal. In a total of five cases, no evidence for a mutation in the genes SRY, ZFY, ZFX, MEA and some anonymous Y-linked sequences was found. In addition to Southern analysis, gene expression of ZFY, ZFX and MEA was found to be normal as well. It is concluded that sex reversal in this condition is due to mutation in a so far unidentified gene which may act secondary to the testis-determining factor (TDF).     

Sex chromosome evolution in fish: the formation of the neo-Y chromosome in Eigenmannia (Gymnotiformes)

Chromosomes of a species of Eigenmannia presenting a X₁X₁X₂X₂:X₁X₂Y sex chromosome system, resulting from a Y-autosome Robertsonian translocation, were analyzed using the C-banding technique, chromomycin A₃ (CMA₃) and mithramycin (MM) staining and in situ digestion by the restriction endonuclease AluI. A comparison of the metacentric Y chromosome of males with the corresponding acrocentrics in females indicated that a C-band-positive, CMA₃/MM-fluorescent and AluI digestion-resistant region had been lost during the process of translocation, resulting in a diminution of heterochromatin in the males. It is hypothesized that the presence of a smaller amount of G+C-rich heterochromatin in the sex chromosomes of the heteromorphic sex when compared with the homomorphic sex may be associated with the sex determination mechanism in this species and may be a more widely occurring phenomenon in fish with differentiated sex chromosomes than was initially thought. Received: 1 April 1999; in revised form: 16 October 1999 / Accepted: 4 December 1999     

Searching for sex‐reversals to explain population demography and the evolution of sex chromosomes

Sex determination can be purely genetic (as in mammals and birds), purely environmental (as in many reptiles), or genetic but reversible by environmental factors during a sensitive period in life, as in many fish and amphibians ( Wallace et al. 1999 ; Baroiller et al. 2009a ; Stelkens & Wedekind 2010 ). Such environmental sex reversal (ESR) can be induced, for example, by temperature changes or by exposure to hormone‐active substances. ESR has long been recognized as a means to produce more profitable single‐sex cultures in fish farms ( Cnaani & Levavi‐Sivan 2009 ), but we know very little about its prevalence in the wild. Obviously, induced feminization or masculinization may immediately distort population sex ratios, and distorted sex ratios are indeed reported from some amphibian and fish populations ( Olsen et al. 2006 ; Alho et al. 2008 ; Brykov et al. 2008 ). However, sex ratios can also be skewed by, for example, segregation distorters or sex‐specific mortality. Demonstrating ESR in the wild therefore requires the identification of sex‐linked genetic markers (in the absence of heteromorphic sex chromosomes) followed by comparison of genotypes and phenotypes, or experimental crosses with individuals who seem sex reversed, followed by sexing of offspring after rearing under non‐ESR conditions and at low mortality. In this issue, Alho et al. (2010) investigate the role of ESR in the common frog (Rana temporaria) and a population that has a distorted adult sex ratio. They developed new sex‐linked microsatellite markers and tested wild‐caught male and female adults for potential mismatches between phenotype and genotype. They found a significant proportion of phenotypic males with a female genotype. This suggests environmental masculinization, here with a prevalence of 9%. The authors then tested whether XX males naturally reproduce with XX females. They collected egg clutches and found that some had indeed a primary sex ratio of 100% daughters. Other clutches seemed to result from multi‐male fertilizations of which at least one male had the female genotype. These results suggest that sex‐reversed individuals affect the sex ratio in the following generation. But how relevant is ESR if its prevalence is rather low, and what are the implications of successful reproduction of sex‐reversed individuals in the wild?     

The use of amplified fragment length polymorphism (AFLP) in the isolation of sex-specific markers

Sex identification is a problem in research and conservation. It can often be solved using a DNA test but this is only an option if a sex-specific marker is available. Such markers can be identified using the amplified fragment length polymorphism (AFLP) technique. This is usually a taxonomic method, as it produces a DNA fingerprint of 50-100 PCR bands. However, if male and female AFLP products are compared, sex-specific markers are confined to the heterogametic sex and can rapidly be identified. Once a marker is found, AFLP can be used to sex organisms directly or the marker can be sequenced and a standard PCR test designed.     

Heteromorphic Z and W sex chromosomes in Physalaemus ephippifer (Steindachner, 1864) (Anura, Leiuperidae)

Heteromorphisms between sex chromosomes are rarely found in anurans and sex chromosome differentiation is considered to be a set of recent recurrent events in the evolutionary history of this group. This paper describes for the first time heteromorphic sex chromosomes Z and W in the leiuperid genus Physalaemus. They were found in P. ephippifer, a species of the P. cuvieri group, and corresponded to the eighth pair of its karyotype. The W chromosome differed from the Z chromosome by the presence of an additional segment in the short arm, composed of a distal NOR and an adjacent terminal DAPI-positive C-band. The identification of this sex chromosome pair may help in future investigations into the sex determining genes in the genus Physalaemus.     

Quantifying and analyzing the network basis of genetic complexity

Genotype-to-phenotype maps exhibit complexity. This genetic complexity is mentioned frequently in the literature, but a consistent and quantitative definition is lacking. Here, we derive such a definition and investigate its consequences for model genetic systems. The definition equates genetic complexity with a surplus of genotypic diversity over phenotypic diversity. Applying this definition to ensembles of Boolean network models, we found that the in-degree distribution and the number of periodic attractors produced determine the relative complexity of different topology classes. We found evidence that networks that are difficult to control, or that exhibit a hierarchical structure, are genetically complex. We analyzed the complexity of the cell cycle network of Sacchoromyces cerevisiae and pinpointed genes and interactions that are most important for its high genetic complexity. The rigorous definition of genetic complexity is a tool for unraveling the structure and properties of genotype-to-phenotype maps by enabling the quantitative comparison of the relative complexities of different genetic systems. The definition also allows the identification of specific network elements and subnetworks that have the greatest effects on genetic complexity. Moreover, it suggests ways to engineer biological systems with desired genetic properties.     

Distribution of Atr protein in primary spermatocytes of a mouse chromosomal mutant: a comparison of preparation techniques

In this study, we examined the suitability of a three dimensional preparation technique for studying chromosome behaviour in the first meiotic prophase in the mouse chromosomal mutant T(1;13)H/T(1;13)Wa. To preserve cellular shape, primary spermatocytes were encapsulated in a fibrin clot. Conventionally sedimented prophase nuclei served as controls. Axial elements and lateral synaptonemal complex components were subsequently stained by immunofluorescence and the presence of axial elements at the pachytene stage was highlighted with indirect immunofluorescence against the Atr protein. We compared the distribution of Atr signal in the fibrin-embedded spermatocytes with surface-spread preparations and immunohistochemically stained histological sections of seminiferous tubules. Furthermore, fluorescence in situ hybridisation of the mouse minor satellite DNA was done on fibrin-embedded spermatocytes. The Atr signal is most conspicuous in fibrin-embedded nuclei on unpaired axial elements during pachytene, both for sex chromosomal and for autosomal segments, and expanding from these elements into the surrounding chromatin. Both spread and encapsulated zygotene nuclei with extended axial element formation proved to be positive for Atr. Mid- to late zygotene nuclei were devoid of 3,3′-diaminodibenzene deposition in the histological sections. Highlighting the unpaired axial elements in the small heteromorphic 1¹³H;1¹³Wa bivalent with an Atr signal enabled meiotic analysis of this bivalent to be carried out in a three-dimensional context. Thus, proximity of this bivalent with the sex chromosomes is found more often in three-dimensional preparations than in spread preparations. Furthermore, the development of the Atr signal over the sex chromosomes as pachytene proceeds helps in substaging of this long and heterogeneous meiotic phase, in sedimented but especially in fibrin-encapsulated nuclei. Received: 22 September 1999; in revised form: 20 December 1999 / Accepted: 21 December 1999     

Conceptual problems and scale limitations of defining ecological communities: a critique of the CI concept (Community of Individuals)

Recently, Looijen & van Andel (1999) proposed a new definition of an ecological community by using two criteria: (1) restricting membership by taxonomic relatedness, and (2) defining boundaries by the intersection of the area of population range boundaries. I analyze the implications of their definition and explore the limitations of the approach. Overall, I show this definition to be highly scale-limited, to not encompass many ecological concepts developed for the community level, and to have hidden assumptions that are not met in natural systems. An alternative model of the ecological community is proposed as a contrast, a model based on the community of an individual, in which individuals and interactions are used to develop the larger entity of an ecological community. This alternative model illustrates that the principal problems Looijen & van Andel (1999) discussed about previous community concepts with respect to application to vegetation classification are not ‘problems’ but are characteristics of ecological communities. Any definition of an ecological community must be able to incorporate these characteristics as well as current ecological concepts used at the community level.     

Meiotic sister chromatid cohesion in holocentric sex chromosomes of three heteropteran species is maintained in absence of axial elements

It has been suggested that in species with monocentric chromosomes axial element (AE) components may be responsible for sister chromatid cohesion during meiosis. To test this hypothesis in species with holocentric chromosomes we selected three heteropteran species with different sex-determining mechanisms. We observed in surface-spreads and sections using transmission electron microscopy that the univalent sex chromosomes form neither AEs nor synaptonemal complexes (SCs) during pachytene. We also found that a polyclonal antibody recognizing SCP3/Cor1, a protein present at AEs and SC lateral elements of rodents, labels the autosomal SCs but not AEs or SC stretches corresponding to the sex chromosomes. Cytological analysis of the segregational behaviour of the sex univalents demonstrates that although these chromosomes segregate equationally during anaphase I they never show precocious separation of sister chromatids during late prophase I or metaphase I. These results suggest that AEs are not responsible for sister cohesion in sex chromosomes. The segregational behaviour of these chromosomes during both meiotic divisions also indicates that different achiasmate modes of chromosome association exist in heteropteran species. Received: 22 September 1999; in revised form: 20 December 1999 / Accepted: 21 December 1999     

鸟类性别决定机制及性别鉴定的研究进展

鸟类的性别决定是一个多基因参与的级联调控过程。这一过程受Z染色体连锁的DMRT1基因, W染色体连锁的PKC1W和其它多种因子共同调控。本文综述了性别决定基因及其功能、性别鉴定方法等方面的研究进展。Abstract： Avian sex determination is a multiple gene regulation cascade. Genes such as the Z chromosome-linked DMRT1 gene, W chromosome-linked PKCIW gene and other factors have been demonstrated to be involved in this process. In this paper, we review the recent progress in this field. The investigation of functions of sex determinate genes and methods of sexing birds are discussed here.     

Evolution and Religion: Attitudes of Scottish Bioscience Students to the Teaching of Evolutionary Biology

In a questionnaire-based survey, the proportion of Glasgow University first year biology students who rejected evolution in 2009–2011 was about 7%, similar to the previously reported average figure for 1987–1999. However, by final year, evolution rejection was absent in students who studied evolution beyond first year and 4% among those who did not. Evolution rejection was closely related to accepting a religion-based alternative, whereas acceptance was related to finding the evidence convincing. Although many religious students accepted evolution, 50% of Islamic students were rejecters, compared to 25% of Christians. A question testing acceptance of several scientific propositions showed no evidence that evolution rejecters were generally more skeptical of science than accepters. Rejecters were overall less secure than accepters in their identification of the correct definition for terms related to evolution and creationism, but, surprisingly, more than 10% of final year students chose a Lamarckian definition for Darwinian evolution. Accepters and rejecters responded equally poorly to a question on Darwin’s history, but level 4 was much better. A breakdown of evolution into three components (human origins, macroevolution, and microevolution) found that some evolution rejecters accepted some components, with microevolution having the highest acceptance and human origins the lowest. These findings are discussed in terms of strategies for evolution education and the phenomenon of evolution rejection worldwide.     

Variables influencing the origins of diverse abnormal behaviors in a large sample of captive chimpanzees (Pan troglodytes)

The developmental origin of abnormal behaviors is generally associated with early rearing environments that lack sufficient physical and sensory stimulation. However, other factors should also be considered. A large sample of captive chimpanzees (128 males and 140 females) was surveyed for the presence or absence of 18 abnormal behaviors. Origin variables included the subject's source (zoo, pet, performer, or laboratory), rearing (mother‐ or hand‐reared), and sex. Animals were assessed while held at the Primate Foundation of Arizona, University of Texas M. D. Anderson Cancer Center, or White Sands Research Center. There was a confound among origin variables; more hand‐reared animals than expected were from laboratories. Logistic regression tested the relationship of rearing and source, with sex as a secondary predictor variable, to each of the abnormal behaviors. There was no clear association between any abnormal behavior and source. However, for coprophagy, relative to animals from the laboratory, zoo animals tended to show a higher prevalence, while performers tended to show a lower prevalence (when rearing and sex were controlled). Rocking and self‐sucking were significantly more likely in hand‐reared animals. Coprophagy and depilation of self were significantly more likely in mother‐reared animals. When rearing and source were statistically controlled, the only significant sex difference was a higher prevalence of coprophagy in females and a higher prevalence of rocking in males. In a second, smaller sample of 25 males and 33 females from Southwest Foundation for Biomedical Research, no significant sex association was found for coprophagy, urophagy, rocking, or self‐depilation. In this second sample, coprophagy was also significantly more likely in mother‐reared than hand‐reared subjects. The association of some abnormal behaviors with mother‐rearing suggests that some form of social learning may be involved in the origin of some of these behavior patterns. This indicates that some abnormal behaviors may not be always be indicative of reduced psychological well‐being in captive chimpanzees. Am. J. Primatol. 48:15–29, 1999. © 1999 Wiley‐Liss, Inc.     

Molecular sex identification of stillborn and neonate individuals ("Traufkinder") from the burial site Aegerten

The study reconstructs the sex ratio of 121 stillborn and neonate individuals from the early modern burial site Aegerten, Switzerland. The immature individuals, who were not baptised before death, were buried along the walls of the church. To perform a molecular sex identification, bone samples from the infants were collected from different skeletal elements. Ancient DNA (aDNA) was isolated by a combination of automated phenol/chloroform extraction and precipitation with silica powder. A combination of manuell Chelex extraction and purification kit was also used to perform an extraction. Finally, the aDNA extracts were amplified with a primer system that amplifies a part of the amelogenin gene located on the human sex chromosomes. The morphometrical sex determination of the children suggests a large disproportion of female individuals (about 60%). This finding was compared to PCR-based amplification results. In contrast, the results of the molecular sex identification were a high proportion of male individuals. Looking at these results, it should be noted that the high mortality of male individuals during the last months of pregnancy and during the first month after birth is in accordance with the natural sequence of death also found in recent populations.     

Sex determination in fish: Lessons from the sex-determining gene of the teleost medaka, Oryzias latipes

Although sex determination systems in animals are diverse, sex-determining genes have been identified only in mammals and some invertebrates. Recently, DMY (DM domain gene on the Y chromosome) has been found in the sex-determining region on the Y chromosome of the teleost medaka fish, Oryzias latipes. Functional and expression analyses of DMY show it to be the leading candidate for the male-determining master gene of the medaka. Although some work is required to define DMY as the master sex-determining gene, medaka is expected to be a good experimental animal for investigating the precise mechanisms involved in primary sex determination in non-mammalian vertebrates. In this article, the process of identification of DMY and is summarized and the origins of DMY and sexual development of the medaka's gonads are reviewed. In addition, putative functions of DMY are discussed.     

Reliability test of the visual assessment of cranial traits for sex determination

Interobserver variation in visual evaluation was analyzed for 10 cranial traits in a homogeneous archaeological series. Two observers independently scored cranial traits commonly used for determination of sex. Though determination of sex did not differ significantly for the two observers, individual traits had different levels of interobserver reliability. In addition, indices of relative "maleness" and "femaleness" derived by the two observers differed at statistically significant levels. Because such indices are used in cross-population comparisons of relative gracility and robusticity of diverse samples, these comparisons should be interpreted with caution when more than one investigator has performed an assessment. Most of our instances of interobserver discordance derived from character traits described in subjective terms without accompanying diagrams. Clarity of definition, rather than number of character traits, was found to be critical for effective determination of sex by the visual assessment method. Use of fewer, more precisely defined character traits can improve interobserver reliability.