Fertility Regulation Research Articles

RNA sequencing and functional analysis uncover key long non-coding RNAs involved in regulating pollen fertility during the process of gametocidal action in wheat.

Gametocidal (Gc) chromosomes have been widely utilized in genetic breeding due to their ability to induce chromosomal breakage and eliminate gametes that lack them. Long noncoding RNAs (lncRNAs) have various functional mechanisms in regulating pollen and anther development; however, their regulatory contributions to Gc action are still unknown. Here, we identified 2824 differentially expressed lncRNAs (DE-lncRNAs) from the anther tissues of Triticum aestivum cv. Chinese Spring (CS) and Chinese Spring-Gc 3C chromosome monosomic addition line (CS-3C) through sequencing. In this study, we predicted 161 target mRNAs for 145 DE-lncRNAs, including 104 cis-regulatory, 60 trans-regulatory, and three both cis-regulatory and trans-regulatory manner. Combined with our previous miRNA sequencing data, 241 DE-lncRNAs functioned as potential endogenous target mimics (eTMs) for 84 differentially expressed microRNAs (DE-miRNAs, including 12 novel miRNAs). The results of transient transformation in tobacco leaves indicated that L006278 could bind to MTCONS_00006277, which encoded a calcineurin CBL-interacting protein kinase 19-like, and suppress its expression. Furthermore, L117735 could function as an eTM for tae-miR9657b-3p, and L056972 could function as an eTM for gc-m2240-5p. To explore the function of lncRNAs in the process of Gc action, we transformed L006278, an up-regulated lncRNA in CS-3C, into rice to analyze its effect on pollen fertility. Overexpression of L006278 led to a reduction in rice pollen fertility. Overall, our findings indicate that lncRNAs can contribute to the regulation of pollen fertility during the process of Gc action by regulating the expression levels of target mRNAs and acting as eTMs for certain key miRNAs.

Production of hydroxyapatite utilizing calcium and phosphorus in sewage sludge incineration ash

Sewage sludge incineration ash (SSIA), enriching the most of phosphorus (P), could be named as promising P source for P recycling and management. The novel strategy including extraction of P from SSIA by acid (HCl), purification and separation to obtain Ca liquid and Al-P liquid that could produce hydroxyapatite was investigated in this study. The P acid leaching rate was 97.9 % from SSIA at 0.2 mol/L HCl for 2 h. In P purification, P combined with Fe and Al to form precipitation (Al-P and Fe-P) with the increase of pH (<4.0). At a pH of around 4.0, the separation of Ca in the liquid phase was maximized, while Al-P and Fe-P precipitated out. Then, the Al-P precipitation could be dissolved in NaOH solution (0.2 mol/L) to separate Fe-P precipitation, that obtaining the Al-P liquid. What’s more, the high P content (P2O5, 25.5 %) in P product could be produced directly under 1.5 Ca/P molar ratio, which could be used in P fertilizer. The heavy metal content in the P product were far below the Chinese, American and EU fertilizer regulation, indicating that its environment safety. The Ca and P elements leached out during the acid leaching process were eventually recombined to form hydroxyapatite by wet-chemical purification. The cost of P recovery from SSIA was $3.06/kg P product demonstrating commercial potential of this strategy.

Genome-wide profiling the epigenetic landscape of histone variant TH2B in murine oocytes and pre-implantation embryos.

The histone variant TH2B, enriched in oocytes, sperm, and early embryos, decreases as embryos differentiate into pre-gastrula stages. Despite its presence, the role of TH2B in epigenetic reprogramming during early embryonic development remains largely under-researched. Our study employed ultra-low-input ChIP-seq (ULI-ChIP) to analyze the genome-wide distribution of TH2B in MII oocytes and early embryos. We found that TH2B is enriched in the chromatin of oocytes and 2-cell stage embryos but becomes less prevalent after the 2-cell stage. Correlation analysis revealed that the TH2B chromatin patterns in sperm and preimplantation embryos are more similar to each other than to those in MII oocytes. Gene ontology (GO) analysis of TH2B-occupied loci linked them to various developmental processes, including oogenesis, fertilization, chromatin modification, and transcription regulation. The study also identified a strong association of TH2B with specific transposable elements (TEs), particularly long terminal repeats (LTRs), which are known to regulate preimplantation development. Additionally, early embryos showed H3K9me3 marks at TH2B-bound loci. TH2B exhibited strong correlations with H2A.Z and H3.3 in the 2-cell and 8-cell stages, a positive association with H3K27Ac and H3K4me3, and a negative correlation with H3K27me3. Allelic reprogramming analysis of TH2B in embryos from C57BL/6J and DBA/2J crosses revealed differential dynamics between maternal and paternal alleles, with a notable paternal bias at the promoter in 2-cell embryos. Thus, TH2B's enrichment in early embryonic stages and its association with key regulatory regions and histone modifications underscore its importance in ZGA and subsequent developmental processes.

Study of the Billings ovulation method for fertility regulation in Lombardy

Study of the Billings ovulation method for fertility regulation in Lombardy

Effects of Microplastics and Organic Fertilizer Regulation on Soil Dissolved Organic Matter Evolution.

Microplastics are pollutants of global concern nowadays. However, the effects of microplastics addition to soil as a carbon source and the combined effects of microplastics and organic fertilizer on soil-dissolved organic matter (DOM) evolution are still unclear. This study focused on the evolution of DOM in soil with the addition of microplastics and investigated the variations in the content and composition of DOM in unfertilized and fertilized soil with different particle sizes of microplastics. It was observed that the TOC concentration of the soil DOM in the treatment with organic fertilizer and microplastics increased more (129.97-161.43 mg kg-1) than that in the treatment with microplastics alone (117.17-131.87 mg kg-1) and was higher than that in the original soil (95.65 mg kg-1). According to the humic acid relative abundance in DOM after 40 days of incubation, the humic acid relative abundance in DOM of the soil samples with microplastics and organic fertilizers addition was found to be higher than that in those with microplastic addition alone, reaching more than 80% in a short time. In conclusion, the TOC concentration of the soil DOM increased with the addition of microplastics, and the increase was more pronounced when organic fertilizers and microplastics were added together. Moreover, the soil humification increased to a higher level in the short term with the combined addition of microplastics and organic fertilizers, which was maintained during the long-term incubation process.

Effect of slow-release mineral fertilizer on the intensity of phosphorus and potassium mobilization in soil

The search for new technologies for effective regulation of yield and soil fertility is an urgent task. The use of slow-release fertilizers can simultaneously solve both problems – reducing fertilizer rates and increasing the mobilization of soil nutrients. In this paper, the mechanism of slow-release fertilizer effect on the mobilization rate of soil nutrients was investigated using mathematical modeling on the data of field trials with winter wheat. It is shown that in the plots with the slowest dissolving fertilizer (azofoska with 20% content of polyvinyl alcohol), the highest efficiency of azofoska was achieved (an increase in yield of 1.3 t/ha and an increase in mobile forms of phosphorus in the soil of 2.5 mg/100 g soil, potassium – 8.8 mg/100 g soil and nitrate nitrogen – 4.7 mg/100 g soil). The results of phosphorus modeling quantitatively confirmed the assumption that the low rate of dissolution of fertilizer enhances the mobilization of nutrients from soil due to the absence of an excess of plant available nutrients. We provide estimates of the polymer added efficiency for a series of polymer-modified fertilizer. It is shown that this form of fertilizer can be applied to effectively reduce the amount of fertilizer required for the planned harvest, simultaneously with the possibility of bringing the soil to a higher level of effective fertility in major nutrient elements.

Extragonadal function of follicle-stimulating hormone: Evidence for a role in endothelial physiology and dysfunction

AimsFollicle-stimulating hormone (FSH) plays a fundamental role in reproduction stimulating ovarian folliculogenesis, Sertoli cells function and spermatogenesis. However, the recent identification of FSH receptor (FSHR) also in extra-gonadal tissues has suggested that FSH activity may not be limited only to fertility regulation, with conflicting results on the possible role of FSH in endothelial cells. The aim of this study was to investigate FSH role on endothelial function in Human Umbilical Vein Endothelial Cells (HUVECs). ResultsEndothelial Nitric oxide synthase (eNOS) expression, eNOS phosphorylation and Nitric Oxide (NO) production resulted increased after the stimulation of HUVEC with recombinant human FSH (rhFSH) at 3.6x103 ng/ml, with increasing Calcium release from intracellular stores. Furthermore, IP3 production increased after rhFSH stimulation despite PTX treatment and NFAT1 was observed prevalently in nucleus.We observed a statistical difference between untreated cells and cells stimulated with 0.36x103 ng/ml and between cells stimulated with 0.36x103 ng/ml and cells stimulated with 1.8x103 ng/ml at 4 and 8 h by Wound healing assay, respectively. Furthermore, a higher cellular permeability was observed in stimulated cells, with atypical VE-cadherin distribution, as well as filamentous actin. ConclusionsOur findings suggest that FSH at high concentrations elicits a signalling that could compromise the endothelial membrane. Indeed, VE-cadherin anomalies may severely affect the endothelial barrier, resulting in an increased membrane permeability. Although NO is an important vasodilatation factor, probably an excessive production could impact on endothelial functionality, partially explaining the increased risk of cardiovascular diseases in menopausal women and men with hypogonadism.

Transgenic Drosophila Expressing Active Human LH Receptor in the Gonads Exhibit a Decreased Fecundity: Towards a Platform to Identify New Orally Active Modulators of Gonadotropin Receptor Activity.

The gonadotropins luteinizing hormone (LH) and follicle-stimulating hormone (FSH) and their receptors are major regulators of reproduction in mammals and are absent in insects. We previously established transgenic Drosophila lines expressing a constitutively active human LH receptor variant (LHRD578Y) and the wild-type receptor (LHRwt; inactive in the absence of an agonist). That study showed that ubiquitously expression of LHRD578Y-but not of LHRwt-resulted in pupal lethality, and targeted expression in midline cells resulted in thorax/bristles defects. To further study the Drosophila model for an in vivo drug screening platform, we investigated here whether expressing LHRD578Y in the fly gonads alters reproduction, as shown in a transgenic mice model. The receptor was expressed in somatic cells of the gonads using the tissue-specific traffic jam-Gal4 driver. Western blot analysis confirmed receptor expression in the ovaries. A fecundity assay indicated that the ectopic expression of LHRD578Y resulted in a decrease in egg laying compared to control flies carrying, but not expressing the transgene (~40% decrease in two independent fly lines, p < 0.001). No significant reduction in the number of laid eggs was seen in flies expressing the LHRWT (<10% decrease compared to non-driven flies, p > 0.05). The decreased egg laying demonstrates a phenotype of the active receptor in the fly gonads, the prime target organs of the gonadotropins in mammals. We suggest that this versatile Drosophila model can be used for the pharmacological search for gonadotropin modulators. This is expected to provide: (a) new mimetic drug candidates (receptor-agonists/signaling-activators) for assisted reproduction treatment, (b) blockers for potential fertility regulation, and (c) leads relevant for the purpose of managing extra gonadotropic reported activities.

Chemogenetic Activation of RFRP Neurons Reduces LH Pulse Frequency in Female but not Male Mice.

The neuropeptide RFRP-3 (RFamide-related peptide-3) is thought to play a role in the negative regulation of fertility. However, the exogenous administration of RFRP-3 yields varying results depending on the dose and route of administration, sex of the subject, and many other variables. Manipulation of in vivo neuronal activity using DREADDs (designer receptor exclusively activated by designer drugs) technology enables investigation of cell type-specific neuronal activation in a manner that better reflects endogenous neuronal activity. To test the effects of RFRP neuronal activation on pulsatile luteinizing hormone (LH) secretion. We generated mice expressing the stimulatory hM3Dq designer receptor exclusively in RFRP cells using 2 different Cre-loxP-mediated approaches: (1) we bred mice to express hM3Dq in all Rfrp-Cre-expressing cells, including some that transiently expressed Rfrp-Cre neonatally (RFRP × hM3Dq mice), and (2) we stereotaxically injected Cre-dependent hM3Dq into the dorsomedial nucleus of RFRP-Cre mice to drive hM3Dq expression exclusively in a subpopulation of adult Rfrp-Cre neurons (RFRP-AAV-hM3Dq mice). We then investigated the effects of acute hM3Dq activation on LH pulse frequency in RFRP × hM3Dq mice, RFRP-AAV-hM3Dq mice, and their respective controls. In both female RFRP × hM3Dq and RFRP-AAV-hM3Dq mice, chemogenetic activation of Cre-driven hM3Dq led to a significant 35% to 50% reduction in LH pulse frequency compared with controls, while no differences in pulse amplitude or mean LH concentration were observed. In marked contrast, RFRP activation did not cause any changes to LH pulse dynamics in male mice. These data show for the first time that activation of neurons that have expressed Rfrp, or of a subset of adult RFRP neurons, can independently suppress LH pulsatility in female, but not male mice.

Elemental composition of post-wildfire biomass ashes and partly burned woody species in Bohemian Switzerland National Park, Czech Republic.

The study explored the post-wildfire elemental composition of parts (wood, bark, branch, cone, trunk, litter, twig, needle, sward, fallow, sapling, etc.) and by-products (biomass ashes, partly burnt parts, and char) of different woody species in the Bohemian Switzerland National Park, Czech Republic, and considered their effects on soils. Multi-elemental analysis of the fire by-products of the woody species was determined with inductively coupled plasma-optical emission spectrometry and mass spectrometry and compared with control biomass samples unaffected by wildfire. Most fire by-products were slightly alkaline, with acidic ashes obtained from blueberry wood. The by-products of the wildfire were characterized by varied total contents of macro (P, Ca, K, Mg, and S), micro (Na, Mn, Fe, Cu, and Zn), and other elements (B, Co, Mo, and V) vital to soil fertility and plant growth. The mean content of macro elements in the biomass ashes was up to 4.16 P, 23.5 Ca, 2.48Mg, 63K, and 5.57 S g kg-1. These values were comparatively lower than published data for ashes obtained under optimized conditions, e.g., those combusted in power generation facilities. Conversely, partly burnt parts-an indication of incomplete combustion-had higher 9.22 P, 79 Ca, and 5.99Mggkg-1 contents in spruce needles than in biomass ashes and the control. Variations in woody species and anthropogenic activities in areas of wildfires produced varied As, Cd, Cr, Ni, and Pb contents above EU fertilizer regulation. Caution in applying biomass ashes from wildfires on fields is required due to risk/toxic elements input from anthropogenic activities. Wildfire effects on the elemental composition of woody species can provide information on plant parts most suitable for biomass ashes for soil and ecosystem safety.

Transcriptional profiling of mucus production in rhesus macaque endocervical cells under hormonal regulation.

Endocervical mucus production is a key regulator of fertility throughout the menstrual cycle. With cycle-dependent variability in mucus quality and quantity, cervical mucus can either facilitate or block sperm ascension into the upper female reproductive tract. This study seeks to identify genes involved in the hormonal regulation of mucus production, modification, and regulation through profiling the transcriptome of endocervical cells from the non-human primate, the rhesus macaque (Macaca mulatta). We treated differentiated primary endocervical cultures with estradiol (E2) and progesterone (P4) to mimic peri-ovulatory and luteal-phase hormonal changes. Using RNA-sequencing, we identified differential expression of gene pathways and mucus producing and modifying genes in cells treated with E2 compared to hormone-free conditions and E2 compared to E2-primed cells treated with P4. We pursued differential gene expression analysis on RNA-sequenced cells. Sequence validation was done using qPCR. Our study identified 158 genes that show significant differential expression in E2-only conditions compared to hormone-free control, and 250 genes that show significant differential expression in P4-treated conditions compared to E2-only conditions. From this list, we found hormone-induced changes in transcriptional profiles for genes across several classes of mucus production, including ion channels and enzymes involved in post-translational mucin modification that have not previously been described as hormonally regulated. Our study is the first to use an in vitro culture system to create an epithelial-cell specific transcriptome of the endocervix. As a result, our study identifies new genes and pathways altered by sex-steroids in cervical mucus production.

Unveiling the abundance and potential impacts of microplastic contamination in commercial organic fertilizers/compost produced from different solid waste.

This study comprehensively investigated the abundance, morphologies, and polymer types of plastics, larger (1-5mm) and smaller (< 1mm) microplastics (MPs), in organic fertilizers using spectroscopic and microscopic methods. MPs abundance varied depending on the type of waste employed. MPs were detected in 80% of the investigated compost samples, while macro/meso plastics were found in only four samples. Compost from mixed municipal solid waste exhibited the highest MPs contamination (23100 ± 3615 items/kg dry weight), whereas compost produced from canteen waste had the lowest contamination (100 ± 65 items/kg dry weight). Smaller MPs were dominant in all samples. The estimated loads of MPs introduced into agricultural soil exceeded the previous studies. Common morphologies observed were sheet, film, fragment, and fiber, while dominant polymer types were polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), and polystyrene (PS). Heavy metals, including Cr, Cu, Ni, and Pb, were identified in association with MPs. Results indicate that the utilization of appropriate waste for composting and upgrading fertilizer regulations is crucial to protect the environment and human health from smaller MPs.

Kisspeptins centrally modulate food intake and locomotor activity in mice independently of gonadal steroids in a sexually dimorphic manner.

Kisspeptins are essential regulators of the reproductive axis, with capacity to potently activate gonadotropin-releasing hormoneneurons, acting also as central conduits for the metabolic regulation of fertility. Recent evidence suggests that kisspeptins per se may also modulate several metabolic parameters, including body weight, food intake or energy expenditure, but their actual roles and site(s) of action remain unclear. We present herein a series of studies addressing the metabolic effects of central and peripheral administration of kisspeptin-10 (Kp-10; 1 nmol and 3 nmol daily, respectively) for 11 days in mice of both sexes. To assess direct metabolic actions of Kp-10 versus those derived indirectly from its capacity to modulate gonadal hormone secretion, kisspeptin effects were tested in adult male and female mice gonadectomized and supplemented with fixed, physiological doses of testosterone or 17β-estradiol, respectively. Central administration of Kp-10 decreased food intake in male mice, especially during the dark phase (~50%), which was accompanied by a reduction in total and nocturnal energy expenditure (~16%) and locomotor activity (~70%). In contrast, opposite patterns were detected in female mice, with an increase in total and nocturnal locomotor activity (>65%), despite no changes in food intake or energy expenditure. These changes were independent of body weight, as no differences were detected in mice of both sexes at the end of Kp-10 treatments. Peripheral administration of Kp-10 failed to alter any of the metabolic parameters analyzed, except for a decrease in locomotor activity in male mice and a subtle increase in 24 h food intake in female mice, denoting a predominant central role of kisspeptins in the control of energy metabolism. Finally, glucose tolerance and insulin sensitivity were not significantly affected by central or peripheral treatment with Kp-10. In conclusion, our data reveal a potential role of kisspeptins in the control of key metabolic parameters, including food intake, energy expenditure and locomotor activity, with a preferential action at central level, which is sex steroid-independent but sexually dimorphic.

Effects of water and fertilizer regulation on soil microbial community, fruit nutrients, and saponin content of Panax notoginseng: A three years field experiment

In pursuit of maximum yield and economic benefits of Panax notoginseng, farmers usually apply excessive fertilizer and irrigation. Which easily induce the decline of Panax notoginseng quality, fertilizer and water use efficiencies and soil health. From 2021–2023, a field experiment was conducted in Dali Village, Luxi County, Honghe Prefecture, Yunnan Province, to study the response of soil microbial community, root nutrient content, and ginsenoside content of Panax notoginseng to different water and fertilizer regulation. The experiment including three irrigation levels (F1: 120 mm, F2: 240 mm, and F3: 360 mm) four fertilization rates(F1: 48 kg ha−1, F2: 72 kg ha−1, F2: 96 kg ha−1, and 120 kg ha−1),and a CK treatment (no fertilizer and no irrigation) The results showed that the average N, P, and K contents across all treatments were greater in 2023 than those in 2021 and 2022. W3F4 treatment significantly increased N absorption. W2F4 treatment significantly increased P and K absorption. W2F4 treatment significantly increased the Panax notoginseng yield, and W3F2 treatment significantly increased the saponin content. The Panax notoginseng yield ranged from 1165 to 3006 kg∙ha−1, with a maximum average yield in 2023. The Re content averaged 81.5 % in 2023 over 2021, and the highest Re content in W3F2 treatment in 2023 was 108.54 mg∙g−1. In 2023, W2F3 treatment had the highest R1 content of 15.34 mg∙g−1, W2F3 treatment had the highest content of Rg1 of 44.65 mg∙g−1, and W3F2 treatment had the highest Rb1 content of 52.52 mg∙g−1. From 2021–2023, the total number of unique OTUs in W1F1 was the largest, out followed by W3F2, W3F4, and W3F1. The treatment that significantly increased the bacterial Chao1 index was W3F1 and W2F2 in 2021, W3F3, W2F1, W2F1 and W2F3 in 2022, and W2F3 and W2F4 in 2023. The Shannon nor Simpson indices were not varied significantly from CK. At the phylum level, W1F3 in 2021 and W3F1 in 2022 and 2023 improved the relative abundance of Proteobacteria, Acidobacteriota At the genus level, W1F1 in 2021 and 2023 improved the relative abundance of Vicinamibacteraceae_norank and Vicinamibacterales_norank. LEfSe analysis of soil microbial communities showed the most divergent species in 2021 with W3F2, W1F1 in 2022, and W2F1 in 2023. Different water and fertilizer treatments affected the total OTU number of bacterial communities and changed the bacterial community structure. W3F1 and W2F3 treatments significantly increased soil bacterial diversity, and soil microbial diversity regulated plants N as well as R1 and Rg1 content, and the quality of Panax notoginseng is related to microbial diversity. N, P, K, and R1 had a significant positive correlation with microbial community diversity (P<0.05). The W1F1 treatment communities were relatively abundant, and the community species showed high similarity in W3F1 and W2F3.

AXDND1 is required to balance spermatogonial commitment and for sperm tail formation in mice and humans

Dynein complexes are large, multi-unit assemblies involved in many biological processes via their critical roles in protein transport and axoneme motility. Using next-generation sequencing of infertile men presenting with low or no sperm in their ejaculates, we identified damaging variants in the dynein-related gene AXDND1. We thus hypothesised that AXDND1 is a critical regulator of male fertility. To test this hypothesis, we produced a knockout mouse model. Axdnd1−/− males were sterile at all ages but presented with an evolving testis phenotype wherein they could undergo one round of histologically replete spermatogenesis followed by a rapid depletion of the seminiferous epithelium. Marker experiments identified a role for AXDND1 in maintaining the balance between differentiation-committed and self-renewing spermatogonial populations, resulting in disproportionate production of differentiating cells in the absence of AXDND1 and increased sperm production during initial spermatogenic waves. Moreover, long-term spermatogonial maintenance in the Axdnd1 knockout was compromised, ultimately leading to catastrophic germ cell loss, destruction of blood–testis barrier integrity and immune cell infiltration. In addition, sperm produced during the first wave of spermatogenesis were immotile due to abnormal axoneme structure, including the presence of ectopic vesicles and abnormalities in outer dense fibres and microtubule doublet structures. Sperm output was additionally compromised by a severe spermiation defect and abnormal sperm individualisation. Collectively these data identify AXDND1 as an atypical dynein complex-related protein with a role in protein/vesicle transport of relevance to spermatogonial function and sperm tail formation in mice and humans. This study underscores the importance of studying the consequences of gene loss-of-function on both the establishment and maintenance of male fertility.

Factors influencing male fertility in Uganda

Male fertility is an important component of population research. Against this backdrop, this paper aims to examine the factors explaining male fertility changes in Uganda. The authors applied the own-children method, as described by Schoumaker, to understand the prevailing male total fertility rate trends and to select datasets for inclusion during the multivariate analysis, using the Uganda demographic health surveys conducted from 2006 to 2016. A multivariable Poisson decomposition regression model was used to examine factors associated with male fertility changes. This model analyzed data from the 2006 and 2016 surveys only and included a total sample size of 7839 male respondents. The outcome variable for the regression model was children ever born (CEB). The regression model showed that differences related to changes in the proportional composition of characteristics contributed 113% to the change in CEB, compared with the differences due to reproductive behavior, with other factors being kept constant. The most important proportional changes that led to meaningful variability in CEB were observed among the following characteristics: occupation, number of current wives, total number of lifetime partners, type of residence, education, and age at first childbirth. Thus, interventions to delay the formation of cohabitation and marriage unions and the onset of childbirth or fatherhood among adolescents and young men below the age of 25 should be prioritized as potent fertility regulation measures. In addition, the focus on eliminating the lack of education and primary education as the highest levels attained among boys and men should be prioritized to boost a rapid male fertility transition in Uganda.

A Quadruple Mutant of OsPRP1 Controls Pollen Fertility by Regulating the Expression of Anther Development-Related Genes in Oryza sativa L.

Plant proline-rich proteins (PRPs) are cell wall proteins that are widely distributed in plants. Previous studies have shown that these proteins play a crucial role in adversity stress processes, but their function in the regulation of pollen fertility in rice remains unknown. In this study, we identified that OsPRP1 contains a Pollen_Ole-e_I allergenic structural domain, obtained the OsPRP1 quadruple mutant named osprp1.1/1.2/1.3/1.4, and observed significant reductions in pollen fertility, seed-setting rates, and the deformation and collapse of microspores during the late stages of pollen development. RNA-Seq analysis indicated the down-regulation of genes involved in anther development in osprp1.1/1.2/1.3/1.4, suggesting that OsPRP1 plays a role in regulating pollen fertility. In conclusion, a loss of function in OsPRP1.1/1.2/1.3/1.4 leads to decreased pollen fertility and seeding rates, which not only expands the functional spectrum of plant PRP genes but also provides new theoretical insight into the mechanism of fertility regulation in rice.

POMC neurons control fertility through differential signaling of MC4R in Kisspeptin neurons.

Inactivating mutations in the melanocortin 4 receptor ( MC4R ) gene cause monogenic obesity. Interestingly, female patients also display various degrees of reproductive disorders, in line with the subfertile phenotype of MC4RKO female mice. However, the cellular mechanisms by which MC4R regulates reproduction are unknown. Kiss1 neurons directly stimulate gonadotropin-releasing hormone (GnRH) release through two distinct populations; the Kiss1 ARH neurons, controlling GnRH pulses, and the sexually dimorphic Kiss1 AVPV/PeN neurons controlling the preovulatory LH surge. Here, we show that Mc4r expressed in Kiss1 neurons is required for fertility in females. In vivo , deletion of Mc4r from Kiss1 neurons in female mice replicates the reproductive impairments of MC4RKO mice without inducing obesity. Conversely, reinsertion of Mc4r in Kiss1 neurons of MC4R null mice restores estrous cyclicity and LH pulsatility without reducing their obese phenotype. In vitro , we dissect the specific action of MC4R on Kiss1 ARH vs Kiss1 AVPV/PeN neurons and show that MC4R activation excites Kiss1 ARH neurons through direct synaptic actions. In contrast, Kiss1 AVPV/PeN neurons are normally inhibited by MC4R activation except under elevated estradiol levels, thus facilitating the activation of Kiss1 AVPV/PeN neurons to induce the LH surge driving ovulation in females. Our findings demonstrate that POMC ARH neurons acting through MC4R, directly regulate reproductive function in females by stimulating the "pulse generator" activity of Kiss1 ARH neurons and restricting the activation of Kiss1 AVPV/PeN neurons to the time of the estradiol-dependent LH surge, and thus unveil a novel pathway of the metabolic regulation of fertility by the melanocortin system.

Cytological Observation and RNA-Seq Analyses Reveal miR9564 and Its Target Associated with Pollen Sterility in Autotetraploid Rice.

Understanding the regulation of autotetraploid sterility is essential for harnessing the strong advantages in genomic buffer capacity, biodiversity, and heterosis of autotetraploid rice. miRNAs play crucial roles in fertility regulation, yet information about their reproductive roles and target genes in tetraploid rice remains limited. Here, we used three tetraploid lines, H1 (fertile), HF (fertile), and LF (sterile), to investigate cytological features and identify factors associated with autotetraploid sterility. LF showed abnormal meiosis, resulting in low pollen fertility and viability, ultimately leading to scarce fertilization and a low-seed setting compared to H1 and HF. RNA-seq revealed 30 miRNA-candidate target pairs related to autotetraploid pollen sterility. These pairs showed opposite expression patterns, with differential expression between fertile lines (H1 and HF) and the sterile line (LF). qRT-PCR confirmed that miR9564, miR528, and miR27874 were highly expressed in the anthers of H1 and HF but not in LF, while opposite results were obtained in their targets (ARPS, M2T, and OsRPC53). Haplotype and expression pattern analyses revealed that ARPS was specifically expressed in lines with the same haplotype of MIR9564 (the precursor of miR9564) as LF. Furthermore, the Dual-GFP assay verified that miR9564 inhibited the fluorescence signal of ARPS-GFP. The over-expression of ARPS significantly decreased the seed setting rate (59.10%) and pollen fertility (50.44%) of neo-tetraploid rice, suggesting that ARPS plays important roles in autotetraploid pollen sterility. This study provides insights into the cytological characteristic and miRNA expression profiles of tetraploid lines with different fertility, shedding light on the role of miRNAs in polyploid rice.

Cellular mechanism underlying leptin-induced anion secretion of rat epididymal epithelial cells.

A large number of studies have shown that leptin plays an important role in the regulation of fertility via the hypothalamus-pituitary-gonad axis. However, its peripheral function in epididymis was still elusive. The purpose of this study was to determine the pro-secretion effect of leptin on the rat epididymal epithelium. In the present study, real-time quantitative polymerase chain reaction, western blot, and immunohistochemical analysis were employed to detect the expression pattern of leptin receptors in rat epididymis. The pro-secretion effect of leptin on epididymal epithelial cells was measured by short-circuit current, and the prostaglandin E2 and cyclic adenosine monophosphate level was evaluated by enzyme-linked immunosorbent assay. We verified that the leptin receptor was located on the epididymal epithelium, with a relatively high expression level in corpus and cauda epididymis. Ussing chamber experiments showed that leptin stimulated a significant rise of the short-circuit current in rat epididymal epithelial cells, which could be abolished by the specific leptin receptor antagonist peptide Allo-aca, or by removing the ambient Cl- and HCO3 -. Furthermore, the leptin-stimulated short-circuit current response could be abrogated by blocking the apical cystic fibrosis transmembrane regulator or the basolateral Na+-K+-2Cl- cotransporter. Our pharmacological experiments manifested that interfering with the prostaglandin H synthase-2-prostaglandin E2-EP2/EP4-adenylate cyclase pathways could significantly blunt the cystic fibrosis transmembrane regulator-mediated anion secretion induced by leptin. The enzyme-linked immunosorbent assay demonstrated that leptin could induce a substantial increase in prostaglandin E2 release and cyclic adenosine monophosphate synthesis of primary cultured rat cauda epididymal epithelial cells. Our data also suggested that JAK2, ERK, and PI3K-dependent phosphorylation may be involved in the activation of prostaglandin H synthase-2 and the subsequent prostaglandin E2 production. The present study demonstrated the pro-secretion function of leptin in rat epididymal epithelium via the activation of cystic fibrosis transmembrane regulator and Na+-K+-2Cl- cotransporter, which was dependent on the paracrine/autocrine prostaglandin E2 stimulated EP2/EP4-adenylate cyclase pathways, and thus contributed to the formation of an appropriate microenvironment essential for sperm maturation.