Sperm Fusion Research Articles

Role of zinc in female reproduction.

Zinc is a critical component in a number of conserved processes that regulate female germ cell growth, fertility, and pregnancy. During follicle development, a sufficient intracellular concentration of zinc in the oocyte maintains meiotic arrest at prophase I until the germ cell is ready to undergo maturation. An adequate supply of zinc is necessary for the oocyte to form a fertilization-competent egg as dietary zinc deficiency or chelation of zinc disrupts maturation and reduces the oocyte quality. Following sperm fusion to the egg to initiate the acrosomal reaction, a quick release of zinc, known as the zinc spark, induces egg activation in addition to facilitating zona pellucida hardening and reducing sperm motility to prevent polyspermy. Symmetric division, proliferation, and differentiation of the preimplantation embryo rely on zinc availability, both during the oocyte development and post-fertilization. Further, the fetal contribution to the placenta, fetal limb growth, and neural tube development are hindered in females challenged with zinc deficiency during pregnancy. In this review, we discuss the role of zinc in germ cell development, fertilization, and pregnancy with a focus on recent studies in mammalian females. We further detail the fundamental zinc-mediated reproductive processes that have only been explored in non-mammalian species and speculate on the role of zinc in similar mechanisms of female mammals. The evidence collected over the last decade highlights the necessity of zinc for normal fertility and healthy pregnancy outcomes, which suggests zinc supplementation should be considered for reproductive age women at risk of zinc deficiency.

Perspective Al-Qur’an and Biology in Human Creation

Abstract. Allah has ordered humans to pay attention to human creation, but the lack of human knowledge and understanding of the correlation of the Qur'an and Biology related to the process of human creation is one of the factors causing a person's lack of faith. This study aims to determine the creation of humans based on the Qur'an that has links with Biological Sciences. This research is library research whose information is obtained through the Qur'an, scientific articles, and biology books. The results of this study are the creation of humans based on the perspective of biology is that God created humans from the fusion of sperm cells and ovum cells. Sperm and ovum cells are part of the human body which are the same components of the human body as components of the soil that have been processed by plants through photosynthesis. Humans were created by God from the fusion of sperm cells from the testes and ovum cells from the ovaries, where the development of the two reproductive organs grows between the sulbi bone (backbone) and breast bone. And humans will be resurrected by God through human bones that are still left in the soil, namely the tailbone.

COMPARISON OF EMBRYO MORPHOKINETICS FOLLOWING ASSISTED SPERM FUSION INSEMINATION AND RESCUE INTRACYTOPLASMIC SPERM INJECTION

In a previous study, assisted sperm fusion insemination (ASFI), which is a fertilization method using a single motile sperm without breaking the oocyte membrane, yielded fertilization rates and blastocyst formation rates similar to those obtained by rescue intracytoplasmic sperm injection (ICSI) (1). In this study, we assessed embryo development following ASFI and rescue ICSI via a time-lapse system.

Cortical Granule Distribution and Expression Pattern of Genes Regulating Cellular Component Size, Morphogenesis, and Potential to Differentiation are Related to Oocyte Developmental Competence and Maturational Capacity In Vivo and In Vitro.

Polyspermia is an adverse phenomenon during mammalian fertilization when more than one sperm fuses with a single oocyte. The egg cell is prepared to prevent polyspermia by, among other ways, producing cortical granules (CGs), which are specialized intracellular structures containing enzymes that aim to harden the zona pellucida and block the fusion of subsequent sperm. This work focused on exploring the expression profile of genes that may be associated with cortical reactions, and evaluated the distribution of CGs in immature oocytes and the peripheral density of CGs in mature oocytes. Oocytes were isolated and then processed for in vitro maturation (IVM). Transcriptomic analysis of genes belonging to five ontological groups has been conducted. Six genes showed increased expression after IVM (ARHGEF2, MAP1B, CXCL12, FN1, DAB2, and SOX9), while the majority of genes decreased expression after IVM. Using CG distribution analysis in immature oocytes, movement towards the cortical zone of the oocyte during meiotic competence acquisition was observed. CGs peripheral density decreased with the rise in meiotic competence during the IVM process. The current results reveal important new insights into the in vitro maturation of oocytes. Our results may serve as a basis for further studies to investigate the cortical reaction of oocytes.

First birth following assisted sperm fusion insemination using sperm bound to zona pellucida.

To report a live birth after transfer of a vitrified-warmed blastocyst produced by assisted sperm fusion insemination (ASFI). Oocyte retrieval and in vitro fertilization (IVF) were performed on a 37-year-old woman. Six hours after IVF, an oocyte exhibited a single polar body and so was defined as an unfertilized oocyte. A motile sperm was collected from the zona pellucida of the unfertilized oocyte by an injection needle. The motile sperm was pressed onto the membrane of the unfertilized oocyte. Two oocytes were matured and subjected to IVF. One of the 2 oocytes exhibited only one polar body and was defined as an unfertilized oocyte at 6h after IVF; this oocyte then was subjected to ASFI. Two pronuclei were observed on the next day and cultured to the blastocyst stage. This embryo achieved blastocyst status and was vitrified on day 5. The resulting vitrified-warmed blastocyst was transferred, resulting in pregnancy and subsequent delivery of a healthy boy. This report describes the first case of a successful birth following transfer of a vitrified-warmed blastocyst produced by ASFI.

Fertilization with human sperm bound to zona pellucida by pressing onto the oocyte membrane.

This study aimed to determine whether fertilization can be obtained by assisted fusion of oocyte and sperm without breaking the oocyte membrane. A total of 79 infertile couples, each with at least one unfertilized oocyte after in vitro fertilization (IVF), were recruited. Sperm collected from the zona pellucida (ZP) were pressed onto the membrane of unfertilized oocytes at either 6h or 24h after IVF, a procedure that we designated as assisted sperm fusion insemination (ASFI). The results of ASFI were compared with those obtained in a previous trial on oocytes in which rescue intracytoplasmic sperm injection (ICSI) was performed at 6h after IVF. Acrosome reaction (AR) rate of sperm bound to ZP, fertilization rate, degeneration rate, and blastocyst formation rate were evaluated. The AR rate of sperm collected from the ZP was significantly higher than that of the motile sperm recovered from around the oocytes but not bound to the ZP after IVF (98.0% vs. 28.6%). ASFI which was performed at 6h after IVF yielded a mean fertilization rate of 73.4% (58/79), a degeneration rate of 0% (0/79) and a blastocyst formation rate of 60.8% (31/51). Rescue ICSI which was performed at 6h after IVF yielded a mean fertilization rate of 70.0% (70/100), a degeneration rate of 4% (4/100) and a blastocyst formation rate of 42.4% (25/59). Binding of sperm to the ZP typically results in AR. ASFI with acrosome-reacted sperm collected from the ZP yielded the fertilization rates similar to those obtained with rescue ICSI.

Maternal control of gamete choice during fertilization.

Sexually reproducing organisms generate male and female haploid gametes, which meet and fuse at fertilization to produce a diploid zygote. The evolutionary process of speciation is achieved and maintained by ensuring that gametes undergo productive fusion only within a species. In animals, hybrids from cross-species fertilization events may develop normally, but are usually sterile (Fitzpatrick, 2004). Metazoan sperm and eggs have several features to ensure that the gametes, which have evolved independently and also in conflict with each other, are competent to undergo fertilization (Firman, 2018). Fertilization is a specific process that is ideally supposed to result in randomized fusion of compatible egg and sperm. Here, I will discuss key processes driven by maternal factors in the egg that dictate earliest stages of gamete recognition, gamete choice and fusion in metazoans.

Embryology of Iris sanguinea Donn ex Horn. and its systematic relationship

Iris sangumea Dorm ex Horn. is a perennial cold-tolerant herbs of the Iris genus of Iridaceae, which has a well-formed, bright color and adaptability. Through understanding the flower bud differentiation, Sporogenesis, gametogenesis, fertilization and embryogenesis of I. sangumea, we developed flowering regulation and control breeding. I. sangumea flower bud differentiation was observed in April for 21 days; in this stage, the early differentiation process was slow, temperature was more than 5 °C, if the temperature is shorter than this, most flower bud will have no differentiation, and turn to vegetative growth; later differentiation is faster and required temperature is from 5 to 8 °C and gradually rose to 10‒12 °C. The temperature in April plays the definitely role in the number of bud differentiation. Three stamina contain tetrasporangiate anthers and a glandular tapetum. The anther wall is composed of four cell layers inclduding the epidermis, the endothecium, one middle layer and the secretory tapetum. The type of anther wall development the dicotyledonous type. After pollen maturation, the anther wall forms a double-layer containing only epidermis and endothecium. Simultaneous cytokinesis is of the continuous type during meiosis of microspore mother cell results in a zygomorphous or tetragonal tetrad. Mature pollen grains are two-celled. I. sangumea’s stylar canal is hollow, open type style, and the gynoecium has three carpels. The ovaries have axial placentas. The ovule is anatropous, bitegmic and crassinucellate. The primary sporogenous cell differentiates directly as the megaspore. The megaspore tetrads are linear, T-shaped or juxtaposed. Generally, the first of the tetrad megaspores at the chalaza becomes functional while the other three degenerate. A mature embryo sac with seven cells and eight nuclei embryo sac is of the polygonum type. For dichogamy, the male gametophyte matures about 2 days earlier than the female gametophyte. The pollen tube entered into the embryo sac for 22–24 h after pollination. The dormancy stage of fertilized ovules is 5‒6 days, and the fusion of sperm and egg nucleus is of the premitotic type. The primary endosperm cell has shorter dormancy of 4‒6 days, and endosperm formation is of the nuclear type.

New advances and future directions in plant polyspermy.

Plants have evolved a battery of mechanisms that potentially act as polyspermy barriers. Supernumerary sperm fusion to one egg cell has consequently long remained a hypothetical concept. The recent discovery that polyspermy in flowering plants is not lethal but generates viable triploid plants is a game changer affecting the field of developmental biology, evolution, and plant breeding. The establishment of protocols to artificially induce polyspermy together with the development of a high-throughput assay to identify and trace polyspermic events in planta now provide powerful tools to unravel mechanisms of polyspermy regulation. These achievements are likely to open new avenues for animal polyspermy research as well, where forward genetic approaches are hampered by the fatal outcome of supernumerary sperm fusion.

FGF10 enhances yak oocyte fertilization competence and subsequent blastocyst quality and regulates the levels of CD9, CD81, DNMT1, and DNMT3B.

The fusion of sperm and oocytes determines the fertilization competence and subsequent development of embryos, which, in turn, can be affected by various proteins and DNA methylation. However, several factors in this whole regulation process remain unknown, especially in yaks. Here, we report that fibroblast growth factor 10 (FGF10) is an important growth factor that can enhance the maturation rate of yak oocytes and the motility of frozen spermatozoa. Subsequent blastocyst quality was also improved by increasing the total cell number and level of pregnancy-associated protein in blastocysts. These effects were significantly high in the group that received the 5 ng/ml FGF10 treatment, during both in vitro maturation (IVM) and capacitation. Our data show that the effects of FGF10 were dose-dependent at vital steps of embryogenesis in vitro. Furthermore, quantitative polymerase chain reaction, western blot analysis, and immunofluorescence demonstrated that the levels of CD9, CD81, DNMT1, and DNMT3B in both mature cumulus-oocyte complexes and capacitated sperms were regulated by FGF10, which was also highly expressed in the group treated with 5 ng/ml FGF10 during both IVM and capacitation. From our present study, we concluded that FGF10 promotes yak oocyte fertilization competence and subsequent blastocyst quality, and could also regulate CD9, CD81, DNMT1, and DNMT3B to optimize sperm-oocyte interactions and DNA methylation during fertilization.

Effects of sperm proteins on fertilization in the female reproductive tract.

Mammalian fertilization that culminates by fusion of the male and female gametes is intricately regulated within the female reproductive tract. To become competent to fertilize an egg, the mammalian spermatozoa that enter the female reproductive tract must undergo a series of physiological changes, including hyperactivation, and capacitation. For reaching full competency, the acrosome, a specialized membrane-bound organelle that covers the anterior part of the sperm head, must undergo an acrosome reaction. For becoming competent to bind an ovum, and to penetrate the zona pellucida and cumulus, many sperm proteins are released in the course of the acrosome reaction. Ultimately, the acrosome binds to the oolemma and fusion of sperm and egg occurs. In this review, we outline current understanding of the roles and effects of some essential sperm proteins and their functions during fertilization in the female reproductive tract.

Chemotactic behavior of egg mitochondria in response to sperm fusion in mice

Mitochondria are the powerhouses of eukaryotic cells and their positioning contributes to fertilization and early developmental processes. We report that sperm fusion triggers Ca2+ oscillations and mitochondrial movement toward fused sperm (mitochondrial chemotaxis) in mouse eggs. Mitochondria functioned in Ca2+ storage and were colocalized with endoplasmic reticulum (ER) during Ca2+ oscillations. Mitochondria then moved toward the fused sperm. Sperm extracts lacking nuclei induced Ca2+ oscillations, but did not promote mitochondrial chemotaxis. Our results suggest that sperm fusion motivates Ca2+ oscillation-independent mitochondrial chemotaxis. This phenomenon indicates that egg mitochondria interact with sperm materials, presumably nuclear substances, and their network tethers egg and sperm nuclei at the early stage of zygote formation.

Sex mosaics in the honeybee: how haplodiploidy makes possible the evolution of novel forms of reproduction in social Hymenoptera.

Hymenoptera are haplodiploid: females arise from fertilized, diploid eggs, while males arise from unfertilized, haploid eggs. The cytogenetic mechanisms underlying haplodiploidy enable remarkable phenomena including female cloning, male cloning and gynandromorphy (sex mosaics). We collected 11 newly emerged putative gynandromorph honeybees from a single colony, assessed the sex of various tissues morphologically and determined the genetic origin (maternal or paternal) of each tissue by genotyping. Ten bees were gynandromorphs with one to three distinct paternal origins. Remarkably, one bee carried no maternal alleles. This bee had female organs throughout, and arose from the fusion of two sperm nuclei. This is the first reported case in the Hymenoptera of sperm fusion resulting in a female, emphasizing the flexibility for social insect reproduction and potentially novel colony-level social structures.

The dualistic origin of human tumors.

Life starts with a zygote, which is formed by the fusion of a haploid sperm and egg. The formation of a blastomere by cleavage division (nuclear division without an increase in cell size) is the first step in embryogenesis, after the formation of the zygote. Blastomeres are responsible for reprogramming the parental genome as a new embryonic genome for generation of the pluripotent stem cells which then differentiate by Waddington’s epigenetic landscape to create a new life. Multiple authors over the past 150 years have proposed that tumors arises from development gone awry at a point within Waddington’s landscape. Recent discoveries showing that differentiated somatic cells can be reprogrammed into induced pluripotent stem cells, and that somatic cell nuclear transfer can be used to successfully clone animals, have fundamentally reshaped our understanding of tumor development and origin. Differentiated somatic cells are plastic and can be induced to dedifferentiate into pluripotent stem cells. Here, I review the evidence that suggests somatic cells may have a previously overlooked endogenous embryonic program that can be activated to dedifferentiate somatic cells into stem cells of various potencies for tumor initiation. Polyploid giant cancer cells (PGCCs) have long been observed in cancer and were thought originally to be nondividing. Contrary to this belief, recent findings show that stress-induced PGCCs divide by endoreplication, which may recapitulate the pattern of cleavage-like division in blastomeres and lead to dedifferentiation of somatic cells by a programmed process known as “the giant cell cycle”, which comprise four distinct but overlapping phases: initiation, self-renewal, termination and stability. Depending on the intensity and type of stress, different levels of dedifferentiation result in the formation of tumors of different grades of malignancy. Based on these results, I propose a unified dualistic model to demonstrate the origin of human tumors. The tenet of this model includes four points, as follows. 1. Tumors originate from a stem cell at a specific developmental hierarchy, which can be achieved by dualistic origin: dedifferentiation of the zygote formed by two haploid gametes (sexual reproduction) via the blastomere during normal development, or transformation from damaged or aged mature somatic cells via a blastomere-like embryonic program (asexual reproduction). 2. Initiation of the tumor begins with a stem cell that has uncoupled the differentiation from the proliferation program which results in stem cell maturation arrest. 3. The developmental hierarchy at which stem cells arrest determines the degree of malignancy: the more primitive the level at which stem cells arrest, the greater the likelihood of the tumor being malignant. 4. Environmental factors and intrinsic genetic or epigenetic alterations represent the risk factors or stressors that facilitate stem cell arrest and somatic cell dedifferentiation. However, they, per se, are not the driving force of tumorigenesis. Thus, the birth of a tumor can be viewed as a triad that originates from a stem cell via dedifferentiation through a blastomere or blastomere-like program, which then differentiates along Waddington’s landscape, and arrests at a developmental hierarchy. Blocking the PGCC-mediated dedifferentiation process and inducing their differentiation may represent a novel alternative approach to eliminate the tumor occurrence and therapeutic resistance.

The Caenorhabditis elegans spe-49 gene is required for fertilization and encodes a sperm-specific transmembrane protein homologous to SPE-42.

Fertilization, the fusion of sperm and oocyte to form a zygote, is the first and arguably the most important cell-cell interaction event in an organism's life. Forward and reverse genetic approaches in the nematode Caenorhabditis elegans have identified many genes that are required for gametogenesis and fertilization and thus are beginning to elucidate the molecular pathways that underlie these processes. We identified an allele of the spe-49 gene in a second filial generation (F2 ) mutagenesis screen for spermatogenesis-defective (spe) mutants. Mutant worms for spe-49 produce sperm that have normal morphology, activate to form ameboid spermatozoa, and can migrate to and maintain their position in the hermaphrodite reproductive tract but fail to fertilize oocytes. This phenotype puts spe-49 in the spe-9 class of late-acting genes that function in sperm at the time of fertilization. We cloned the spe-49 gene through a combination of deficiency mapping, transgenic rescue, and genomic sequencing. spe-49 messenger RNA (mRNA) is enriched in male germ cells, and the complementary DNA (cDNA) encodes a predicted 772-amino-acid six-pass transmembrane protein that is homologous to SPE-42. Indeed, SPE-49 and SPE-42 have identical predicted membrane topology and domain structure, including a large extracellular domain with six conserved cysteine residues, a DC-STAMP domain, and a C-terminal cytoplasmic domain containing a C4-C4 RING finger motif. The presence of two SPE-42 homologs in animal genomes from worms to humans suggests that these proteins are highly conserved components of the molecular apparatus required for the sperm-oocyte recognition, binding, and fusion.

Hetero-fertilization together with failed egg–sperm cell fusion supports single fertilization involved in in vivo haploid induction in maize

In vivo doubled-haploid technology is widely applied in commercial maize breeding programs because of its time-saving and cost-reducing features. The production of maize haploids primarily depends on the use of Stock6-derived haploid inducer lines. Although the gene underlying haploid induction, MTL/ZmPLA1/NLD, was cloned recently, the mechanism of haploid induction is still unknown. Hetero-fertilization can occur via a single fertilization, which provides a means to investigate single-fertilization events by studying the hetero-fertilization phenomenon. In this study, we found that the hetero-fertilization rate increased significantly when female maize lines were first individually crossed with pollen from the inducer CAU5 in dual-pollination experiments 4 h before a second pollination with common lines. We also examined embryogenesis during haploid induction by confocal laser-scanning microscopy and observed single-fertilized ovules, indicating that single fertilization occurred during haploid induction. We therefore postulate that both single fertilization and chromosome elimination contribute to haploid induction in maize. We also propose a scheme for the formation of hetero-fertilized and haploid kernels. Our results provide an efficient approach to identify hetero-fertilized kernels for research on interactions between embryo and endosperm.

Effects of Kapok Seed Extract(Ceiba pentandra Gaertn) as Contraceptive Agent to the Quality of the Spermatozoa in Domestic Cat (Felis domestica)

Contraception is a way to prevent the fusion of mature egg cell and sperm during the intercourse, so the fertilization will not occur. Some contraceptive methods in small animals have been evolved. This research aimed to determine the effects of kapok seed (Ceiba pentandra Gaertn) extract as contraception agent to the quality of spermatozoain domestic cats. The animals were divided into four group with each group consisted of six male cats. Group I, II, III was designated as the treatment groups and IV as the control group. The cats were given the emulsion of kapok seeds extract orally with concentration of 0,1% b/v, 0,71 mg/kg BB, 0,36 mg/kg BB and 0,21 mg/kg BB doses. This study used Randomized Complete Random Design (RAL) design from sperm quality data and used ANOVA statistic test. The result showed that the concentration of sperm decreased in the treatment groups compared with the control group. Statistically, the sperm concentration decreased significantly (P < 0,05) between the control group and treatment groups. The average motility in low and medium doses was above 40% while it was below than 40% in the high doses group. Based on the results, the granting of kapok seed extract inhibited the viability of spermatozoa of cats with significant results (P < 0,05). Sperm abnormalities in this study was obtained from the control group with percentage average of 1 % , while it was significantly increased in the treatment groups marked by abnormalities in the shape of the head and tail of the spermatozoa. These results suggested that kapok seed extract can be used as one of the contraceptive agents in domestic cat (Felis domestica) because it can decrease the sperm quality significantly.

What Can Stem Cell Models Tell Us About Human Germ Cell Biology?

Fusion of sperm and egg generates a totipotent zygote that develops into a whole organism. Accordingly, the "immortal" germline transmits genetic and epigenetic information to subsequent generations with consequences for human health and disease. In mammals, primordial germ cells (PGCs) originate from peri-gastrulation embryos. While early human embryos are inaccessible for research, in vitro model systems using pluripotent stem cells have provided critical insights into human PGC specification, which differs from that in mice. This might stem from significant differences in early embryogenesis at the morphological and molecular levels, including pluripotency networks. Here, we discuss recent advances and experimental systems used to study mammalian germ cell development. We also highlight key aspects of germ cell disorders, as well as mitochondrial and potentially epigenetic inheritance in humans.

Fertilisation of polar nuclei and formation of early endosperms in Dendrobium catenatum: evidence for the second fertilisation in Orchidaceae

Whether the second fertilisation, i.e. fertilisation of polar nuclei, or fusion of the second sperm with polar nuclei occurs in Orchidaceae has long been controversial because of lack of evidence. In the present study, we observed fusion and fertilisation of polar nuclei and formation of early endosperms in the orchid Dendrobium catenatum Lindl., by using a resin-embedded section technique. As the product of the second fertilisation, the primary endosperm nucleus (fertilised polar nuclei) can last until the global embryo stage, indicating that initiation of endosperm development and that of embryo development were fully asynchronous. The present study demonstrated the occurrence of the second fertilisation in D. catenatum by providing lines of new evidence.

Triparental plants provide direct evidence for polyspermy induced polyploidy

It is considered an inviolable principle that sexually reproducing organisms have no more than two parents and fertilization of an egg by multiple sperm (polyspermy) is lethal in many eukaryotes. In flowering plants polyspermy has remained a hypothetical concept, due to the lack of tools to unambiguously identify and trace this event. We established a high-throughput polyspermy detection assay, which uncovered that supernumerary sperm fusion does occur in planta and can generate viable polyploid offspring. Moreover, polyspermy can give rise to seedlings with one mother and two fathers, challenging the bi-organismal concept of parentage. The polyspermy derived triploids are taller and produce bigger organs than plants resulting from a regular monospermic fertilization. In addition, we demonstrate the hybridization potential of polyspermy by instantly combining three different Arabidopsis accessions in one zygote. Our results provide direct evidence for polyspermy as a route towards polyploidy, which is considered a major plant speciation mechanism.