Male Sexual Development Research Articles

In vitro obtainment of stem-like cells from gubernaculum testis biopsies of cryptorchid pediatric patients

Testicular descent is a crucial event in male sexual development, and alterations in this process during gestation can lead to reduced fertility in adulthood. Cryptorchidism, i.e., failure of one or both testicles to descend into the scrotum, is one of the most common birth defects and represents a principal cause of infertility in adulthood. Therefore, identifying effective approaches for preserving fertility in childhood is of primary importance. In this context, the key role played by the gubernaculum testis during the placement of the testes in the scrotal bursa emerges. Given its close affinity to testicular tissue and its richness in mesenchymal cells, our prime aim is to characterize this para-testicular tissue to explore its potential ability to differentiate into testicular cells for fertility preservation. The first step of our task is represented by the present study that aimed to obtain in vitro stem-like cells starting from gubernaculum testis biopsies of four pediatric patients affected by cryptorchidism, aiming to differentiate them into testicular functioning cells. Our results show that the obtainment of aggregates with stem features is not dependent on the age of the patients and, therefore, not even on the damage suffered by the testis during its stay in the abdomen. This study opens the possibility of extending this approach to older patients, offering a new potential approach to support their fertility potential.

The impact of mosaic loss of the Y chromosome (mLOY) in men of advanced age.

The Y chromosome has long been considered to be a "genetic wasteland" harboring only few genes essentially involved in male sex development and spermatogenesis. However, the discovery of mosaic loss of the Y chromosome (mLOY) in older men has led to revisiting of the potential impact of the Y chromosome on health and the pathophysiological processes of multiple diseases such as cancer, Alzheimer's disease and cardiovascular disease. Hence, developing more sensitive techniques for the detection of mLOY has become an emergent concern. In this article, we present a comprehensive review of the literature regarding mLOY. Additionally, we discuss the emerging discoveries concerning mLOY as well as the underlying mechanisms promoting disease in men of advanced age.

Epigenetic control of SOX9 gene by the histone acetyltransferase P300 in human Sertoli cells

BackgroundThe transcription factor SOX9 is a key regulator of male sexual development and Sertoli cell differentiation. Altered SOX9 expression has been implicated in the pathogenesis of disorders of sexual development (DSD) in mammals. However, limited information exists regarding the epigenetic mechanisms governing its transcriptional control during sexual development. MethodsThis study employed real-time PCR (qPCR), immunofluorescence (IIF), and chromatin immunoprecipitation (ChIP) assays to investigate the epigenetic mechanisms associated with SOX9 gene transcriptional control in human and mouse Sertoli cell lines. To identify the specific epigenetic enzymes involved in SOX9 epigenetic control, functional assays using siRNAs for P300, GCN5, and WDR5 were performed. ResultsThe transcriptional activation of SOX9 was associated with selective deposition of active histone modifications, such as H3K4me3 and H3K27ac, at its enhancer and promoter regions. Importantly, the histone acetyltransferase P300 was found to be significantly enriched at the SOX9 enhancers, co-localizing with the H3K27ac and the SOX9 transcription factor. Silencing of P300 led to decreased SOX9 expression and reduced H3K27ac levels at the eSR-A and e-ALDI enhancers, demonstrating the crucial role of P300-mediated histone acetylation in SOX9 transcriptional activation. Interestingly, another histone lysine acetyltransferases like GNC5 and methyltransferases as the Trithorax/COMPASS-like may also have a relevant role in male sexual differentiation. ConclusionsHistone acetylation by P300 at SOX9 enhancers, is a key mechanism governing the transcriptional control of this essential regulator of male sexual development. These findings provide important insights into the epigenetic basis of sexual differentiation and the potential pathogenesis of DSDs.

PCR amplification, sequencing, and in silico analysis of holandric genes TSPYand SRY in Nili Ravi buffalo

The present work has been designed to study the evolutionary perspectives of two important holandric genes, namely, testis-specific protein, Y-encoded (TSPY), and sex determining region, Y-encoded (SRY) in Nili Ravi buffalo. These Y-chromosomal genes are responsible for male sex development. The evolutionary aspects of these genes have not been studied in the Nili Ravi buffaloes from India. A study was carried out on the peripheral blood samples of three adult Nili Ravi bulls belonging to university farm. DNA was extracted using the phenol-chloroform isoamyl alcohol (PCI) technique and reported primer sequences were used to amplify the genes (partially) using a thermocycler. Three biological samples were analyzed using Sanger sequencing and were aligned using Clustal Omega. Separate phylogenetic trees for the genes were constructed using MEGA7 software. The selection pressure analysis and evolutionary divergence were individually studied for each of the genes. The current study reports the partial TSPY and SRY gene sequences of Indian Nili Ravi buffalo. Besides, the molecular evolution analysis indicates that the genes have experienced some purifying selection during the evolutionary process. Molecular characterization of these holandric genes (TSPY and SRY) in Nili Ravi will find applications in sex determination in Nili Ravi and other buffaloes.

Formation of Male Sex Tilapia Larvae (Oreochromis niloticus) Using Natural Methyltestosterone Hormone

Aims: Obtaining a productive tilapia, by changing the sex of the tilapia larvae into male monosex. This was achieved by determining the effect of the duration of administrating the natural methyltestosterone (MT) hormone on the male sex of tilapia using the Immersion method.
 Methodology: The study was conducted at the Northern Region Marine and Fisheries Service Branch, Wanayasa Freshwater Fish Seed Development Center, Purwakarta Regency, West Java Province, Indonesia. The experimental design used a completely randomized design (CRD) with 5 treatments and 3 replications. The study used the Immersion method where untreated tilapia and the four hormones tilapia. The treatments are as follows:
 A = untreated tilapia
 B = tilapia treated in 17a-MT (5h-100mg/L)
 C = tilapia treated in MT (5h-350mg/L)
 D = tilapia treated in MT (10h-350mg/L)
 E = tilapia treated in MT (15h-350mg/L)
 Results: Tilapia larvae used were aged 6 days after hatching and reared for 40 days, then the male and SR sex were observed. The results of this study showed that the duration of immersion of tilapia larvae using natural MT significantly affected the male tilapia which resulted in the highest male percentage at 15 h of immersion of 80.2%, then followed by 10-h immersion of 75.6% with the same SR value of about 90%.
 Conclusion: Administration of natural MT using cow testis meal by immersion method at different durations significantly affects male sexual development in 6-day-old tilapia larvae. Based on the obtained data analyses, the percentage of male tilapia in the 40-day experimental study shows the most effective duration in treatment E (15 h) with a measure of 350 mg/l, which is 80.2%. Further studies regarding the relationship between sex change and the growth of sex-changed fish as an effect of immersion duration.

Long-Term Outcome of Horizontal Root Fractures in Permanent Teeth: A Retrospective Cohort Study

IntroductionHorizontal root fracture (HRF) is a complex traumatic dental injury that affects the pulp, dentin, cementum, and periodontal ligament. This retrospective cohort study evaluated treatment outcomes in permanent teeth with HRF. MethodsWe analyzed clinical and radiographic data from a dental trauma center (2006–2022). Permanent teeth with HRF with a follow up of ≥12 weeks were considered for outcome assessment (defined as clinical normalcy and radiographic healing at the fracture line). Prognostic factors were identified through multivariable logistic regression analyses (P value ≤ .05). Results125 teeth from 103 patients were included. After a median follow-up of 79 weeks, the overall favorable outcome was 92%. This includes teeth that received emergency splinting/repositioning at baseline (62.2%) and those that received subsequent endodontic intervention for the coronal fragment (baseline: 85%; subsequent follow-ups: 91.8%). Being male and incomplete root development were both significantly associated with a better outcome of splinting/repositioning (OR = 2.58; 95% CI, 1.06-6.24 and OR = 4.37; 95% CI, 1.16-16.41, respectively) and a reduced likelihood of requiring endodontic treatment (OR = 0.44; 95% CI, 0.20-0.96 and OR = 0.24; 95% CI, 0.08-0.76, respectively). Treatment delays surpassing one week were significantly associated with an increased likelihood of requiring endodontic treatment compared to timely presentations within 24 hours (OR = 3.06; 95% CI, 1.07-8.77; P value < .05). ConclusionWith timely diagnosis and treatment, and close monitoring, HRF cases can achieve a 92% favorable outcome. Male sex and incomplete root development correlate with improved baseline outcomes and a reduced need for endodontic treatment. Conversely, delayed presentation increases the likelihood of requiring endodontic intervention.

Identification and functional verification of Y-chromosome-specific gene typo-gyf in Bactrocera dorsalis.

Genes on the Y chromosome play important roles in male sex determination and development. The identification of Y-chromosome-specific genes not only provides a theoretical basis for the study of male reproductive development, but also offers genetic control targets for agricultural pests. However, Y-chromosome genes are rarely characterized due to their high repeatability and high heterochromatinization, especially in the oriental fruit fly. In this study, 1011 Y-chromosome-specific candidate sequences were screened from 2 to 4h Bactrocera dorsalis embryo datasets with the chromosome quotient method, 6 of which were identified as Y-chromosome-specific sequences by polymerase chain reaction, including typo-gyf, a 19126-bp DNA sequence containing a 575-amino acid open reading frame. Testicular deformation and a significant reduction in sperm number were observed after typo-gyf knockdown with RNA interference in embryos. After typo-gyf knockout with clustered regularly interspaced palindromic repeats (CRISPR) / CRISPR-associated protein 9 in the embryonic stage, the sex ratio of the emergent adults was unbalanced, with far more females than males. A genotype analysis of these females with the Y-chromosome gene MoY revealed no sex reversal. Typo-gyf knockout led to the death of XY individuals in the embryonic stage. We conclude that typo-gyf is an essential gene for male survival, and is also involved in testicular development and spermatogenesis. The identification of typo-gyf and its functional verification provide insight into the roles of Y-chromosome genes in male development.

The testis, eunuchs, and testosterone: a historical review over the ages and around the world.

Testosterone therapy for men with testosterone deficiency is widely used, yet remains controversial. The rich and fascinating history of the testicles, including human castration, provides a valuable perspective on this important topic. This study reviewed the history of testosterone from antiquity to the modern day. Primary sources consisted of books and relevant articles, augmented by a MEDLINE search using the key words "testis," "testicles," "castration," "eunuchs," "testosterone," and "testicular function." An early scientific observation was that castration reduced sexual development and activity, originating with domestication of animals approximately 10 000years ago. Human castration appears in ancient Egyptian mythology more than 4000years ago, in Greek mythology from 8th century BCE, and in the Bible. The history of eunuchs in China spanned 2000years, beginning with the Hsia dynasty (2205-1766BCE). The concept that the testicles produced some factor responsible for male sexual development and behavior was thus known throughout the world since the beginning of recorded history. Testosterone was isolated and synthesized in 1935 and was soon available as a treatment. Multiple benefits of testosterone therapy were apparent by 1940. Recent large, controlled testosterone studies have conclusively demonstrated sexual and general health benefits, with a strong safety profile. Testosterone has been a known substance for <1% of the historical timeline, yet knowledge that the testes were responsible for male sexual development and behavior has been known since the beginning of recorded history. Today, modern evidence has demonstrated the importance of normal levels of testosterone for general health as well as sexual function and desire. Yet, testosterone therapy remains controversial. We believe this historical review provides a helpful perspective on this age-old issue.

Plant reproduction: Ancient origins of male germline differentiation

Despite the wide diversity in male sexual development across land plants, new work reveals the conservation of a heterodimer of transcription factors as master regulators of the male germline.

Data-driven networking of global transcriptomics and male sexual development in the main malaria vector, Anopheles funestus

Deaths from malaria remain staggering despite global support that drives research into new territories. One major gap is our understanding of the sexual biological aspects of the male mosquito, which maintain the vector population solidity. Although Anopheles funestus s.s. is an extremely efficient African vector, little is known about the network between its sexual physiology and gene expression. The Culicidae male’s sexual maturity involves a suite of physiological changes, such as genitalia rotation that is necessary for successful mating to occur. We show that mating success is guided by genes and physiological plasticity. Transcriptome analysis between newly emerged males (immature) versus males with rotating genitalia (maturing) provides insight into possible molecular mechanisms regulating male sexual behaviour. Putative transcripts that were associated with male sexual maturation were identified and validated. The discovery of the functions of these transcripts could lead to identifying potential targets for innovative vector control interventions, and mosquito population suppression.

Iraqi experience in the assessment of anti-Müllerian hormone utilization in clinical practice: An overview

Abstract Anti-Müllerian hormone (AMH) is a dimeric glycoprotein, first recognized due to its action in male sex development. In women, it is synthesized by granulosa cells, pre-antral, and antral follicles; its main function in the ovary appears to be the inhibition of the early steps of folliculogenesis. Measurement of serum AMH made the assessment of ovarian reserve possible for more than 30 years. Its use has been expanded for many clinical implications, including assisted reproductive techniques, menopause, reproductive dysfunctions, the degree of ovarian damage/toxicity, and others. Numerous studies executed in this field concluded that AMH declines with aging; it has a positive correlation with oocyte yield in assisted reproduction; it is upregulated in polycystic ovarian syndrome and decreases in ovarian surgery and cancer therapy. This review aims to highlight the Iraqi experience in the implication of AMH assessment in clinical practice. We reviewed the PubMed database regarding the most recent literature in the assessment of AMH in Iraqi Clinical Practice; studies in polycystic ovarian syndrome (PCOS) patients were excluded. The review results show the importance of AMH assessment in many clinical fields especially for the evaluation of ovarian reserve of an infertile woman either to begin treatment strategists or follow-up patients and oncofertility counseling; yet studies must be encouraged in Iraq about it.

Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice.

Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense-point-mutation mouse model (LhcgrW495X ) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild-type SLCs are transplanted into LhcgrW495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein-split PEmax system via lentivirus successfully corrects the mutation in SLCs from LhcgrW495X mice ex vivo. Gene-corrected SLCs from LhcgrW495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene-corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in LhcgrW495X mice. Altogether, these results suggest that PE-based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system.

An overview on androgen-mediated actions in skeletal muscle and adipose tissue

Androgens are a class of steroid hormones primarily associated with male sexual development and physiology, but exert pleiotropic effects in either sex. They have a crucial role in various physiological processes, including the regulation of skeletal muscle and adipose tissue homeostasis. The effects of androgens are mainly mediated through the androgen receptor (AR), a ligand-activated nuclear receptor expressed in both tissues. In skeletal muscle, androgens via AR exert a multitude of effects, ranging from increased muscle mass and strength, to the regulation of muscle fiber type composition, contraction and metabolic functions. In adipose tissue, androgens influence several processes including proliferation, fat distribution, and metabolism but they display depot-specific and organism-specific effects which differ in certain context. This review further explores the potential mechanisms underlying androgen-AR signaling in skeletal muscle and adipose tissue. Understanding the roles of androgens and their receptor in skeletal muscle and adipose tissue is essential for elucidating their contributions to physiological processes, disease conditions, and potential therapeutic interventions.

Identification of Male Sex-Related Genes Regulated by SDHB in Macrobrachium nipponense Based on Transcriptome Analysis after an RNAi Knockdown.

The oriental river prawn (Macrobrachium nipponense) is a commercially important species in Asia. A previous study showed that the succinate dehydrogenase complex iron sulfur subunit B (SDHB) gene participates in testes development in this species through its effect on the expression of the insulin-like androgenic gland hormone gene. This study knocked-down the Mn-SDHB genes in M. nipponense using RNAi. A transcriptome analysis of the androgenic gland and testes was then performed to discover the male sex-related genes regulated by SDHB and investigate the mechanism of male sexual development in this species. More than 16,623 unigenes were discovered in each sample generated. In the androgenic gland, most of the differentially expressed genes were enriched in the hypertrophic cardiomyopathy pathway, while in the testes, they were enriched in the citrate cycle pathway. In addition, after Mn-SDHB knockdown, five genes were found to be downregulated in the androgenic gland in a series of biological processes associated with phosphorylated carbohydrate synthesis and transformations in the glycolysis/gluconeogenesis pathway. Moreover, a total of nine male sex-related genes were identified including Pro-resilin, insulin-like androgenic gland hormone, Protein mono-ADP-ribosyltransferase PAPR11, DNAJC2, C-type Lectin-1, Tyrosine-protein kinase Yes, Vigilin, and Sperm motility kinase Y-like, demonstrating the regulatory effects of Mn-SDHB, and providing a reference for the further study of the mechanisms of male development in M. nipponense.

A Candidate Bacterial-Type Amino Acid Decarboxylase Is Essential for Male Gamete Exflagellation and Mosquito Transmission of the Malaria Parasite.

A frequent side effect of chemotherapy against malaria parasite blood infections is a dramatic induction of the sexual blood stages, thereby enhancing the risk of future malaria transmissions. The polyamine biosynthesis pathway has been suggested as a candidate target for transmission-blocking anti-malarial drug development. Herein, we describe the role of a bacterial-type amino acid decarboxylase (AAD) in the life cycle of the malaria model parasite Plasmodium yoelii. Hallmarks of AAD include a conserved catalytic lysine residue and high-level homology to arginine/lysine/ornithine decarboxylases of pathogenic bacteria. By targeted gene deletion, we show that AAD plays an essential role in the exflagellation of microgametes, resulting in complete absence of sporozoites in the mosquito vector. These data highlight the central role of the biosysthesis of polyamines in the final steps of male gamete sexual development of the malaria parasite and, hence, onward transmission to mosquitoes.

A selective androgen receptor modulator, S4, displays robust anti-cancer activity on hepatocellular cancer cells by negatively regulating PI3K/AKT/mTOR signalling pathway

Hepatocellular carcinoma (HCC) is a major global health problem that often correlates with poor prognosis. Due to the insufficient therapy options with limited benefits, it is crucial to identify new therapeutic approaches to overcome HCC. One of the vital signaling pathways in organ homeostasis and male sexual development is Androgen Receptor (AR) signaling. Its activity affects several genes that contribute to cancer characteristics and have essential roles in cell cycle progression, proliferation, angiogenesis, and metastasis. AR signaling has been shown to be misregulated in many cancers, including HCC, suggesting that it might contribute to hepatocarcinogenesis. Targeting AR signaling using anti-androgens, AR inhibitors, or AR-degrading molecules is a powerful and promising strategy to defeat HCC. In this study, AR signaling was targeted by a novel Selective Androgen Receptor Modulator (SARM), S4, in HCC cells to evaluate its potential anti-cancer effect. To date, S4 activity in cancer has not been demonstrated, and our data unrevealed that S4 significantly impaired HCC growth, migration, proliferation, and induced apoptosis through inhibiting PI3K/AKT/mTOR signaling. Since PI3K/AKT/mTOR signaling is frequently activated in HCC and contributes to its aggressiveness and poor prognosis, its negative regulation by the downregulation of critical components via S4 was a prominent finding. Further studies are necessary to investigate the S4 action mechanism and anti-tumorigenic capacity in in-vivo.

Low serum anti-Müllerian hormone is associated with semen quality in infertile men and not influenced by vitamin D supplementation

BackgroundAnti-Müllerian hormone (AMH) is released by testicular Sertoli cells and of great importance during fetal male sexual development, but less is known about the role of circulating AMH during adulthood. In vitro studies have shown that vitamin D may induce AMH transcription, but a controlled trial investigating the possible effect of vitamin D on serum AMH has not been conducted in men.MethodsA single-center, double-blinded, randomized placebo-controlled clinical trial (NCT01304927) conducted in Copenhagen, Denmark. A total of 307 infertile men were included and randomly assigned (1:1) to a single dose of 300,000 IU cholecalciferol followed by 1400 IU cholecalciferol + 500 mg of calcium daily (n = 151) or placebo (n = 156) for 150 days. Difference in serum AMH was a predefined secondary endpoint. Explorative outcomes were associations between serum AMH and gonadal function in infertile men. The primary endpoint was difference in semen quality and has previously been published.ResultsInfertile men in the lowest AMH tertile had significantly lower sperm concentration (∆T3-1 16 mill/mL (228%); P < 0.001), sperm count (∆T3-1 55 million (262%); P < 0.001), motile sperm count (∆T3-1 28 million (255%); P < 0.001), progressive motile sperm count (∆T3-1 18 million (300%); P < 0.001), testis size (∆T3-1 2.7 mL (16%); P < 0.001), serum inhibin B (∆T3-1 72 pg/mL (59%); P < 0.001), inhibin B/FSH ratio (∆T3-1 48 (145%); P < 0.001), and higher FSH (∆T3-1 2.6 (38%); P < 0.001) than the tertile of infertile men with highest serum AMH. Vitamin D supplementation had no effect on serum AMH compared with placebo treatment.ConclusionsIn infertile men, low serum AMH is associated with severely impaired gonadal function illustrated by poor semen quality and lower testosterone/LH ratio. Serum AMH in infertile men was not influenced by vitamin D supplementation.

IAG Regulates the Expression of Cytoskeletal Protein-Encoding Genes in Shrimp Testis.

Insulin-like androgenic gland hormone (IAG) is the master regulator of sexual differentiation and testis development in male crustaceans. However, the molecular mechanism on how IAG functions during testis development is still largely unknown. Here, the transcriptional changes were analyzed in the testes of shrimp after LvIAG knockdown in Litopenaeus vannamei. Differential expression analysis identified 111 differentially expressed genes (DEGs), including 48 upregulated DEGs and 63 downregulated DEGs, in testes of shrimp after LvIAG knockdown. Gene ontology (GO) analysis showed that these DEGs were apparently enriched in cytoskeleton-related GO items. Gene function analysis showed that genes enriched in these GO items mainly encoded actin, myosin, and heat shock protein. Interestingly, these genes were all downregulated in testis after LvIAG knockdown, which was confirmed by qRT-PCR detection. Furthermore, injection of LvIAG protein that was recombinantly expressed in insect cells upregulated the expression levels of these genes. The present study revealed that shrimp IAG might function in testis development through regulating the expression of cytoskeletal protein-encoding genes, which would provide new insights into understanding the functional mechanisms of IAG on male sexual development of crustaceans.

Germline Mutations in Steroid Metabolizing Enzymes: A Focus on Steroid Transforming Aldo-Keto Reductases.

Steroid hormones synchronize a variety of functions throughout all stages of life. Importantly, steroid hormone-transforming enzymes are ultimately responsible for the regulation of these potent signaling molecules. Germline mutations that cause dysfunction in these enzymes cause a variety of endocrine disorders. Mutations in SRD5A2, HSD17B3, and HSD3B2 genes that lead to disordered sexual development, salt wasting, and other severe disorders provide a glimpse of the impacts of mutations in steroid hormone transforming enzymes. In a departure from these established examples, this review examines disease-associated germline coding mutations in steroid-transforming members of the human aldo-keto reductase (AKR) superfamily. We consider two main categories of missense mutations: those resulting from nonsynonymous single nucleotide polymorphisms (nsSNPs) and cases resulting from familial inherited base pair substitutions. We found mutations in human AKR1C genes that disrupt androgen metabolism, which can affect male sexual development and exacerbate prostate cancer and polycystic ovary syndrome (PCOS). Others may be disease causal in the AKR1D1 gene that is responsible for bile acid deficiency. However, given the extensive roles of AKRs in steroid metabolism, we predict that with expanding publicly available data and analysis tools, there is still much to be uncovered regarding germline AKR mutations in disease.

Regulators of male and female sexual development are critical for the transmission of a malaria parasite

Malaria transmission to mosquitoes requires a developmental switch in asexually dividing blood-stage parasites to sexual reproduction. In Plasmodium berghei, the transcription factor AP2-G is required and sufficient for this switch, but how a particular sex is determined in a haploid parasite remains unknown. Using a global screen of barcoded mutants, we here identify genes essential for the formation of either male or female sexual forms and validate their importance for transmission. High-resolution single-cell transcriptomics of ten mutant parasites portrays the developmental bifurcation and reveals a regulatory cascade of putative gene functions in the determination and subsequent differentiation of each sex. A male-determining gene with a LOTUS/OST-HTH domain as well as the protein interactors of a female-determining zinc-finger protein indicate that germ-granule-like ribonucleoprotein complexes complement transcriptional processes in the regulation of both male and female development of a malaria parasite.