Sex Determination Research Articles

Determination of Sex Using Frontal Sinus Morphometry with CBCT

Determination of Sex Using Frontal Sinus Morphometry with CBCT

Sex Determination by Using Discriminant Function Analyses from the Northeastern-Thai Occipital Bones

Sex Determination by Using Discriminant Function Analyses from the Northeastern-Thai Occipital Bones

Transcriptomic analysis of sexually dimorphic cypris larvae of the rhizocephalan barnacle Peltogasterella gracilis

Peltogasterella gracilis (Cirripedia: Rhizocephala), a crustacean parasite of hermit crabs, displays genotypic sex determination. Its larvae are planktonic, and female larvae settle on the host. Subsequently, the females control the host's behavior by spreading a root-like structure called “the interna” within the host's body, and form sacs containing eggs called “the externae” outside the host's body. On the other hand, male larvae settle on immature externae and become dwarf males. The cypris larvae of P. gracilis show sexual dimorphism in size and morphology. However, there is no understanding of the molecular mechanisms underlying the sexual dimorphism observed at the larval stage. Here, we conducted a transcriptome analysis and compared the expression of genes in male and female cyprids to better understand their sexual differentiation and settlement processes. A total of 2870 differentially expressed transcripts, comprising 456 female- and 2414 male-biased transcripts were identified. Among the male-biased ones, ionotropic glutamate receptor-, heat shock protein-, acetylcholine-, and homeobox-, cuticle-related transcripts were included. Additionally, 29 gene ontology terms were associated with the sex-specific traits. The present study improves our understanding of sex determination, sexual differentiation, and settlement processes of rhizocephalans.

Reference-based genome assembly and comparative genomics of Calamus Brandisii Becc. for unveiling sex-specific genes for early gender detection.

Calamus brandisii Becc. is an endangered rattan species indigenous to the Western Ghats of India and used in the furniture and handicraft industries. However, its dioecious nature and longer flowering time pose challenges for conservation efforts. Developing markers for early gender detection in seedlings is crucial for maintaining viable populations for in-situ and ex-situ conservation. Currently, no sex chromosomes or gender-specific genes have been reported in the species. We report the first comprehensive comparative genomics study between the male and female genomes of C. brandisii to identify polymorphisms and potential genes for gender determination. Reference-based assembly was conducted and the male and female genomes were predicted to contain 43,810 and 50,493 protein-coding genes respectively. The haploid genome size was ∼691Mb and ∼884Mb for male and female genomes respectively. Comparative analysis revealed significant genetic variation between the two genomes including 619,776 SNPs, 73,659 InDels, 212,123 Structural variants (SVs) and 305 copy number variations (CNVs). A total of 5 male-specific and 11 female-specific genes linked to the sex determining region was predicted. The genomic variants identified between the two genomes could be used in development of markers for early gender identification in C. brandisii for restoration programs. The gender-specific genes identified in this study also provide new insights into the mechanisms of sex determination and differentiation in rattans.

Performances of Different Classification Algorithms in Sex Determination from First Cervical Vertebra Measurements

Performances of Different Classification Algorithms in Sex Determination from First Cervical Vertebra Measurements

Identifying the sex chromosome and sex determination genes in the cattle tick, Rhipicephalus (Boophilus) microplus.

Rhipicephalus (Boophilus) microplus is globally one of the most economically important ectoparasites of cattle costing the agriculture industry billions of dollars annually. Resistance to chemical control measures has prompted the development of novel methods of control. Recent advancements in genetic control measures for human and other animal vectors have utilized sex determination research to manipulate sex ratios, which have shown promising results in mosquitoes namely Aedes aegypti and Anopheles stephensi. Here, we use R. (B.) microplus as a model to provide foundational research to allow similar avenues of investigation in ticks using R. (B.) microplus as a model. Karyotypes for R. (B.) microplus show an XX:XO sex determining system with the largest chromosome being the sex chromosome. Using flow cytometric methods, the size of the sex chromosome was estimated at 526.91 Mb. All measures to identify the sex chromosome within the cattle tick genome assembly associated sex chromosomal characteristics to two chromosomes. This discrepancy between the assembly and karyotypes of the tick led to generating a new genome assembly with a single adult male specimen. The two chromosomes in question aligned with a single scaffold within the new genome that had a length of 513.29 Mb and was the first time the sex chromosome was identified in an Ixodid genome assembly.

The extraordinary Atlantic silverside and me

Abstract My career is inextricably linked to the remarkable characteristics and astonishing insights provided by a simple yet elegant species, the Atlantic silverside, Menidia menidia. My initial field and laboratory studies provided the first demonstration of temperature-dependent sex determination (TSD) and its adaptive significance in fish, as well as the first empirical confirmation of frequency-dependent sex ratio selection. This early work then led to the discovery of countergradient variation in growth, evolutionary trade-offs associated with growth, pioneered the study of fisheries-induced evolution, and furthered our understanding of the spatial scale of local adaptation in marine species. Most of these topics are now vigorous areas of research encompassing numerous species and systems. Here I describe how these unforeseen discoveries were made and led to other findings on bluefish (Pomatomus saltatrix) and striped bass (Morone saxatilis). An interest in leadership during my early years was later rekindled as I took on administrative roles. It all began with an intriguing observation stemming from my MS thesis that I was compelled to follow. My career illustrates the value of exploring deeply the natural history of lesser-known yet easy-to-study species and to follow unexpected observations even if peripheral to a preconceived plan of research.

Fem-1 Gene of Chinese White Pine Beetle (Dendroctonus armandi): Function and Response to Environmental Treatments.

Dendroctonus armandi (Tsai and Li) (Coleoptera: Curculionidae: Scolytinae) is regarded as the most destructive forest pest in the Qinling and Bashan Mountains of China. The sex determination of Dendroctonus armandi plays a significant role in the reproduction of its population. In recent years, the role of the fem-1 gene in sex determination in other insects has been reported. However, the function and expression of the fem-1 gene in Dendroctonus armandi remain uncertain. In this study, three fem-1 genes were cloned and characterized. These were named Dafem-1A, Dafem-1B, and Dafem-1C, respectively. The expression levels of these three Dafem-1 genes vary at different stages of development and between the sexes. In response to different environmental treatments, including temperature, nutrients, terpenoids, and feeding duration, significant differences were observed between the three Dafem-1 genes at different developmental stages and between males and females. Furthermore, injection of double-stranded RNA (dsRNA) targeting the expressions of the Dafem-1A, Dafem-1B, and Dafem-1C genes resulted in increased mortality, deformity, and decreased emergence rates, as well as an imbalance in the sex ratio. Following the interference with Dafem-1A and Dafem-1C, no notable difference was observed in the expression of the Dafem-1B gene. Similarly, after the interference with the Dafem-1B gene, no significant difference was evident in the expression levels of the Dafem-1A and Dafem-1C genes. However, the interference of either the Dafem-1A or Dafem-1C gene results in the downregulation of the other gene. The aforementioned results demonstrate that the Dafem-1A, Dafem-1B, and Dafem-1C genes play a pivotal role in the regulation of life development and sex determination. Furthermore, it can be concluded that external factors such as temperature, nutrition, terpenoids, and feeding have a significant impact on the expression levels of the Dafem-1A, Dafem-1B, and Dafem-1C genes. This provides a crucial theoretical foundation for further elucidating the sex determination mechanism of Dendroctonus armandi.

The expressions of nr0b1/2 genes in the Chinese giant salamander and their response to estradiol benzoate/17α-methyltestosterone treatment

Previously, the nr0b1 and nr0b2 genes have been identified in various animals and found to be closely associated with sex determination. In order to investigate the regulatory roles of nr0b1 and nr0b2 genes in the sex determination of Chinese giant salamander (CGS, Andrias davidianus), we obtained the full-length cDNA sequences of nr0b1 and nr0b2 through cloning, determined the characteristics of their tissue expressions and the responses of nr0b1 and nr0b2 to estradiol benzoate (EB) and 17α-methyltestosterone (MT) by using quantitative real-time PCR (qRT-PCR), and evaluated gonad development through histological sections. The full-lengths of cDNA of nr0b1/2 in C GS are separately 1278bp and 789bp. They have complete open reading frames (ORF) of 864bp and 774bp and encode 287 and 257 amino acids respectively. The 3' untranslated regions (UTR) are 276bp and 15bp, and the 5' UTR of nr0b1 is 138bp. In the 3' UTR of nr0b1, a putative polyadenylation signal (AATAAA) is identified. Tissue distribution analysis indicates that nr0b1 is mainly expressed in the pituitary, kidney, and testes, with significant differences between females (ZW-F) and males (ZZ-M) expression in the testes. On the other hand, nr0b2 is mainly expressed in the liver, followed by gonadal tissues. After immersion and feeding with EB and MT, EB's promotion on nr0b1 expression in CGS ovaries and testes is shown, by contrast MT only has a positive effect on nr0b2 in CGS testes. Histological sections show that after EB treatment, both genetic females (ZW-F) and sex-reversed females (ZZ-F) of CGS contain oogonia, indicating that EB can cause sex reversal in CGS; however, after MT treatment, no cellular morphological changes are observed either in genetic females (ZW-F) or in genetic males (ZW-M), indicating that this concentration cannot induce a shift in the sex ratio of CGS. These findings lay the foundation for further investigating the molecular mechanisms by which nr0b1/2 influence the differentiation and maintenance of CGS gonads.

From fibers, to flowering, to metabolites: unlocking hemp (Cannabis sativa L.) potential with the guidance of novel discoveries and tools.

Cannabis sativa L. is an ancient crop whose agricultural adoption has been interrupted to prevent the use of marijuana as psychoactive drug. Nevertheless, hemp - the Cannabis sativa type with low concentrations of intoxicating Δ9-tetrahydrocannabinoid - is experiencing resurged interest thanks to loosened cultivation restrictions and its potential as multipurpose bio-based crop. In fact, hemp has valuable applications, including production of medicines from its non-intoxicating cannabinoids, food, medical, and industrial uses of its seed oil rich in poly-unsaturated fatty acids, and production of fibers for textiles and industry from its stems. Recently, several hemp genomic and genetic resources have been developed, allowing for a significant expansion of the genetic knowledge on major hemp traits, as cannabinoids, oil, and fibers synthesis, and regulation of flowering and sex determination. Still, hemp is an under-improved crop, whose advancement will depend on the ability to expand and collectively use the novel resources available in light of the fast advancements in bioinformatics and plant phenotyping technologies. This review discusses on the current genetic and genomic knowledge on the most important hemp traits, and provides a perspective on how to further expand such knowledge and tackle hemp improvement with the most up-to-date tools for plant and hemp research.

Automated High-Throughput Biological Sex Identification from Archeological Human Dental Enamel Using Targeted Proteomics.

Biological sex is key information for archeological and forensic studies, which can be determined by proteomics. However, the lack of a standardized approach for fast and accurate sex identification currently limits the reach of proteomics applications. Here, we introduce a streamlined mass spectrometry (MS)-based workflow for the determination of biological sex using human dental enamel. Our approach builds on a minimally invasive sampling strategy by acid etching, a rapid online liquid chromatography (LC) gradient coupled to a high-resolution parallel reaction monitoring (PRM) assay allowing for a throughput of 200 samples per day (SPD) with high quantitative performance enabling confident identification of both males and females. Additionally, we developed a streamlined data analysis pipeline and integrated it into a Shiny interface for ease of use. The method was first developed and optimized using modern teeth and then validated in an independent set of deciduous teeth of known sex. Finally, the assay was successfully applied to archeological material, enabling the analysis of over 300 individuals. We demonstrate unprecedented performance and scalability, speeding up MS analysis by 10-fold compared to conventional proteomics-based sex identification methods. This work paves the way for large-scale archeological or forensic studies enabling the investigation of entire populations rather than focusing on individual high-profile specimens. Data are available via ProteomeXchange with the identifier PXD049326.

Single-nucleus multiomics reveals the gene-regulatory networks underlying sex determination of murine primordial germ cells.

Accurate specification of female and male germ cells during embryonic development is critical for sexual reproduction. Primordial germ cells (PGCs) are the bipotential precursors of mature gametes that commit to an oogenic or spermatogenic fate in response to sex-determining cues from the fetal gonad. The critical processes required for PGCs to integrate and respond to signals from the somatic environment in gonads are not understood. In this study, we developed the first single-nucleus multiomics map of chromatin accessibility and gene expression during murine PGC development in both XX and XY embryos. Profiling of cell-type specific transcriptomes and regions of open chromatin from the same cell captured the molecular signatures and gene networks underlying PGC sex determination. Joint RNA and ATAC data for single PGCs resolved previously unreported PGC subpopulations and cataloged a multimodal reference atlas of differentiating PGC clusters. We discovered that regulatory element accessibility precedes gene expression during PGC development, suggesting that changes in chromatin accessibility may prime PGC lineage commitment prior to differentiation. Similarly, we found that sexual dimorphism in chromatin accessibility and gene expression increased temporally in PGCs. Combining single-nucleus sequencing data, we computationally mapped the cohort of transcription factors that regulate the expression of sexually dimorphic genes in PGCs. For example, the gene regulatory networks of XX PGCs are enriched for the transcription factors, TFAP2c, TCFL5, GATA2, MGA, NR6A1, TBX4, and ZFX. Sex-specific enrichment of the forkhead-box and POU6 families of transcription factors was also observed in XY PGCs. Finally, we determined the temporal expression patterns of WNT, BMP, and RA signaling during PGC sex determination, and our discovery analyses identified potentially new cell communication pathways between supporting cells and PGCs. Our results illustrate the diversity of factors involved in programming PGCs towards a sex-specific fate.

An explanation for the prevalence of XY over ZW sex determination in species derived from hermaphroditism

The many independent transitions from hermaphroditism to separate sexes (dioecy) in flowering plants and some animal clades must often have involved the emergence of a heterogametic sex-determining locus, the basis of XY and ZW sex determination (i.e., male and female heterogamety). Current estimates indicate that XY sex determination is much more frequent than ZW, but the reasons for this asymmetry are unclear. One proposition is that separate sexes evolve through the invasion of sterility mutations at closely linked loci, in which case XY sex determination evolves if the initial male sterility mutation is fully recessive. Alternatively, dioecy may evolve via the gradual divergence of male and female phenotypes, but the genetic basis of such divergence and its connection to XY and ZW systems remain poorly understood. Using mathematical modeling, we show how dioecy with XY or ZW sex determination can emerge from the joint evolution of resource allocation to male and female function with its genetic architecture. Our model reveals that whether XY or ZW sex determination evolves depends on the trade-off between allocation to male and female function, and on the mating system of the ancestral hermaphrodites, with selection for female specialization or inbreeding avoidance both favoring XY sex determination. Together, our results cast light on an important but poorly understood path from hermaphroditism to dioecy, and provide an adaptive hypothesis for the preponderance of XY systems. Beyond sex and sex determination, our model shows how ecology can influence the way selection shapes the genetic architecture of polymorphic traits.

Selective loss of Y chromosomes in lung adenocarcinoma modulates the tumor immune environment through cancer/testis antigens.

There is increasing recognition that the sex chromosomes, X and Y, play an important role in health and disease that goes beyond the determination of biological sex. Loss of the Y chromosome (LOY) in blood, which occurs naturally in aging men, has been found to be a driver of cardiac fibrosis and heart failure mortality. LOY also occurs in most solid tumors in males and is often associated with worse survival, suggesting that LOY may give tumor cells a growth or survival advantage. We analyzed LOY in lung adenocarcinoma (LUAD) using both bulk and single-cell expression data and found evidence suggesting that LOY affects the tumor immune environment by altering cancer/testis antigen expression and consequently facilitating tumor immune evasion. Analyzing immunotherapy data, we show that LOY and changes in expression of particular cancer/testis antigens are associated with response to pembrolizumab treatment and outcome, providing a new and powerful biomarker for predicting immunotherapy response in LUAD tumors in males.

Two phases of gonadal sex differentiation in zebrafish with ZZ/ZW sex determination system

Zebrafish sex chromosomes have been identified in the wild Nadia (NA) strain, and its sex determination belongs to the female-heterogametic ZZ/ZW system. Here, we investigate the correlation between ZZ/ZW sex chromosomes in the NA strain with sex-related factors, and sort out the complicated process of sex determination in zebrafish. Two phases exist during zebrafish sex differentiation. In the first phase, ZW gonads differentiate into juvenile ovary while ZZ gonads remain indifferent. In the second phase, ZW gonads either continue ovary development or undergo female-to-male transition, while ZZ gonads undergo direct male development. The W chromosome may contribute to the first phase while the abundance of germ cells and other factors may be involved in the second phase of sex differentiation in zebrafish.

Sex, gender, and stroke recovery: Functional limitations and inpatient care needs in Canadian and European survivors.

Stroke is a leading cause of long-term disability among survivors. Past literature already investigated the biological sex differences in stroke outcome; still limited work on gender differences is published. Therefore, the study aimed at investigating whether biological sex and sociocultural gender of survivors play a role as determinants of disability and quality of life among stroke survivors across Europe and Canada. Data were gathered from the European Health Information Survey (EHIS, n = 316,333) and Canadian Community Health Survey (CCHS, n = 127,462) data sets. Main outcomes of interest were disability, assessed through evaluating the impairment of Activities of Daily Living (ADL) and Instrumental Activities of Daily Living (iADL), and inpatient care needs, such as hospitalization or institutionalization. Multivariate logistic regression models were utilized to identify factors independently associated with outcomes. Federated analysis was conducted for cross-country comparisons. Data were adjusted for the country-specific Gender Inequality Index (GII), with higher score corresponding to more gender inequality toward females. Female survivors showed greater impairments in iADL (odds ratio [OR] = 1.73, 95% confidence interval [CI]:1.53-1.96) and ADL (OR = 1.25, 95% CI: 1.09-1.44), without a corresponding increase in inpatient care needs. Socioeconomic factors such as marital status and income level were significant predictors of disability, with low income and being single/divorced associated with higher risks. The impact of sex was more pronounced in countries with higher GII, indicating the influence of gender inequality on stroke outcomes. The findings highlight the significant impact of biological sex and gender-related social determinants on post stroke disability, with female sex and unfavorable socioeconomic conditions being associated with worse outcomes.

Spatio-temporal human snow leopard (Panthera uncia) conflicts and mitigation measures in Baltistan - free-livestock grazing pastoral areas

Apex predators are essential for maintaining ecosystem balance by controlling biotic communities. The spatio-temporal grazing of livestock in pastoral areas can pose threats to endangered carnivores and their prey resulting in resource competition, conflicts and increased disease transmission. Our research aimed to assess the spatio-temporal distribution of snow leopards and livestock determine factors causing snow leopard-livestock conflicts identify seasonal conflict zones through public claims and interviews and evaluate conservation benefits and challenges. In this study, camera traps were used in 20 villages of Baltistan from November to April resulting in 720 snow leopard sightings and 8277 livestock events. Snow leopards preyed on 195 livestock in the 2022–2023 period resulting in an estimated loss of $225,004.51 USD including sheeps, goats, cows and yaks.We utilized the Generalized Linear Regression Model to examine snow leopard attacks finding that the timing of attacks was not significant. Proximity to predator-proofed corrals, traditional corrals, bite marks and distance to the nearest village or shepherd huts were positively correlated with attacks, while small stock body size was not significant. Through 110 public interviews we discovered that awareness campaigns and avoidance strategies successfully mitigated human-snow leopard conflicts. In conclusion, our results found that livestock attacks occur in pastures during both day and night with the size and sex of the livestock affecting the likelihood of attacks. Snow leopard attacks were linked to increased grazing and poorly constructed corrals. Snow leopards targeted vulnerable areas of their prey including the neck, mouth, ears and anal region. Compensation claims and field visits were used to identify conflict areas and public perception. In the future research should prioritize the use of camera traps to study prey and snow leopard density, grazing effects, food habits using genetic tools and sex determination.

Targeted Metabolites and Transcriptome Analysis Uncover the Putative Role of Auxin in Floral Sex Determination in Litchi chinensis Sonn.

Litchi exhibits a large number of flowers, many flowering batches, and an inconsistent ratio of male and female flowers, frequently leading to a low fruit-setting rate. Floral sexual differentiation is a crucial phase in perennial trees to ensure optimal fruit production. However, the mechanism behind floral differentiation remains unclear. The objective of the study was to identify the role of auxin in floral differentiation at the transcriptional level. The results showed that the ratio of female flowers treated with naphthalene acetic acid (NAA) was significantly lower than that of the control stage (M0/F0). The levels of endogenous auxin and auxin metabolites were measured in male and female flowers at different stages of development. It was found that the levels of IAA, IAA-Glu, IAA-Asp, and IAA-Ala were significantly higher in male flowers compared to female flowers. Next-generation sequencing and modeling were employed to perform an in-depth transcriptome analysis on all flower buds in litchi 'Feizixiao' cultivars (Litchi chinensis Sonn.). Plant hormones were found to exert a significant impact on the litchi flowering process and flower proliferation. Specifically, a majority of differentially expressed genes (DEGs) related to the auxin pathway were noticeably increased during male flower bud differentiation. The current findings will enhance our comprehension of the process and control mechanism of litchi floral sexual differentiation. It also offers a theoretical foundation for implementing strategies to regulate flowering and enhance fruit production in litchi cultivation.

BIOLOGICAL ASPECTS OF Ameivula nigrigula (SQUAMATA: TEIIDAE): FEEDING HABIT, SEXUAL DIMORPHISM, REPRODUCTION AND MATURATION OF FEMALES

Life history information plays a crucial role in comprehending the evolutionary processes that shape ecological patterns within species, but there is limited information available on the reproductive and other aspects of tropical Squamata. In this study, we present a comprehensive analysis of dietetic and morphometric data, specifically related to the size and sex determination of Ameivula nigrigula, aiming to investigate if there is sexual dimorphism and identify the onset of sexual maturity in female individuals. By thoroughly examining the stomach contents, we observed that the majority of specimens retain food remnants, with a prevalence of small prey items such as Hymenoptera and Isoptera. Our findings reveal a lack of sexual dimorphism in terms of body size, indicating that males and females possess similar dimensions and proportions. Moreover, our investigations suggest that different color patterns do not correspond to distinct stages of female development, as chromatic variations are observed in both juvenile and ovigerous females. Consequently, dorsal coloration may not serve as a reliable indicator of sexual maturity. The observed chromatic variation is likely a form of polymorphism within the population, lacking the potential to determine life stages. Furthermore, our examination of female fertility reveals the presence of one to two eggs per female, consistent with findings in other Teiidae species.

Early sex identification by leaflet distance in plantlets of Cycas revoluta

In dioecious plants, the process of sex determination usually occurs during the reproductive stage. However, it can be challenging to determine the sex of long-lived plants with long generation times. This is true for Cycas revoluta, which produces toxic seeds in female plants, leading to a preference for male plants in public green spaces. In this study we aim to identify a morphological trait that can be used to distinguish between the sexes in C. revoluta at a very early stage of plant growth. We sampled C. revoluta seedlings/plantlets at three different early growth stages and identified the sex of individuals by PCR amplification of a male-specific molecular marker. On the same plants, we measured morphological traits, including the distance between leaflets on the leaf rachis, perimeter, area, number of indents, and leaf complexity, and evaluated their correlation with the sex of the individuals. Among all measured traits, the medium distance between leaflets on the leaf rachis was found to be a sexually dimorphic morphological trait in Cycas plantlets in all three growth stages. In particular, the distance between leaflets on the leaf rachis in the upper part of the leaf in the first stage revealed no overlap between the sexes. Whenever available, morphological traits for sex differentiation in greenhouses and plant nurseries can represent a rapid and economic tool when coupled with automatized image recognition systems.