Oocyte Size Research Articles

HOP-1 Presenilin Deficiency Causes a Late-Onset Notch Signaling Phenotype That Affects Adult Germline Function in Caenorhabditis elegans.

Functionally redundant genes present a puzzle as to their evolutionary preservation, and offer an interesting opportunity for molecular specialization. In Caenorhabditis elegans, either one of two presenilin genes (sel-12 or hop-1) facilitate Notch activation, providing the catalytic subunit for the γ secretase proteolytic enzyme complex. For all known Notch signaling events, sel-12 can mediate Notch activation, so the conservation of hop-1 remains a mystery. Here, we uncover a novel "late-onset" germline Notch phenotype in which HOP-1-deficient worms fail to maintain proliferating germline stem cells during adulthood. Either SEL-12 or HOP-1 presenilin can impart sufficient Notch signaling for the establishment and expansion of the germline, but maintenance of an adult stem cell pool relies exclusively on HOP-1-mediated Notch signaling. We also show that HOP-1 is necessary for maximum fecundity and reproductive span. The low-fecundity phenotype of hop-1 mutants can be phenocopied by switching off glp-1/Notch function during the last stage of larval development. We propose that at the end of larval development, dual presenilin usage switches exclusively to HOP-1, perhaps offering opportunities for differential regulation of the germline during adulthood. Additional defects in oocyte size and production rate in hop-1 and glp-1 mutants indicate that the process of oogenesis is compromised when germline Notch signaling is switched off. We calculate that in wild-type adults, as much as 86% of cells derived from the stem cell pool function to support oogenesis. This work suggests that an important role for Notch signaling in the adult germline is to furnish a large and continuous supply of nurse cells to support the efficiency of oogenesis.

Oocyte size distribution reveals ovary development strategy, number and relative size of egg batches in lumpfish (Cyclopterus lumpus)

The reproductive biology of fishes impact many other components of their life history, and can influence their vulnerability to fisheries, therefore for more informed management, a good understanding is essential. For lumpfish (Cyclopterus lumpus), a semi-pelagic species found across the north-Atlantic and targeted by fishers for their roe, comprehensive knowledge of this aspect of their life history is still lacking. Through a combination of regular sampling from scientific surveys and fisheries and modern methodology in fish reproductive biology, we investigated the ovary development of lumpfish throughout vitellogenesis. The results showed that ovaries of lumpfish had a wide range of oocyte sizes and that lumpfish are a determinate, batch spawner with ovary development taking at least 8 months. They spawn a maximum of two batches per season with a similar number of eggs in each batch. Unusually for a determinate batch spawner, the two batches were easily distinguished within the ovary prior to ovulation. Average egg size ranged from between 2050 and 2500 µm, with larger fish having larger eggs, and the egg diameter of the second batch being on average 1.6% smaller than the first. Lumpfish were documented as spawning over a 4-month period, but it is likely that spawning occurs over a greater period. A macroscopic and oocyte size frequency distribution (OSFD) scale for lumpfish is presented which can be used for future studies of lumpfish.

Quantitative mRNA Imaging with Dual Channel qFIT Probes to Monitor Distribution and Degree of Hybridization.

Fluorogenic oligonucleotide probes facilitate the detection and localization of RNA targets within cells. However, quantitative measurements of mRNA abundance are difficult when fluorescence signaling is based on intensity changes because a high concentration of unbound probes cannot be distinguished from a low concentration of target-bound probes. Here, we introduce qFIT (quantitative forced intercalation) probes that allow the detection both of probe-target complexes and of unbound probes on separate, independent channels. A surrogate nucleobase based on thiazole orange (TO) probes the hybridization status. The second channel involves a nonresponsive near-IR dye, which serves as a reporter of concentration. We show that the undesirable perturbation of the hybridization reporter TO is avoided when the near-IR dye Cy7 is connected by means of short triazole linkages in an ≥18 nucleotides distance. We used the qFIT probes to localize and quantify oskar mRNA in fixed egg chambers of wild-type and mutant Drosophila melanogaster by wash-free fluorescence in situ hybridization. The measurements revealed a relative 400-fold enrichment of oskar within a 3000 μm3 large volume at the posterior pole of stage 8-9 oocytes, which peaked at a remarkably high 1.8 μM local concentration inside 0.075 μm3 volume units. We discuss detection limits and show that the number of oskar mRNA molecules per oocyte is independent of the oocyte size, which suggests that the final levels are attained already during the onset of oskar localization at stage 8.

Oocyte recruitment and fecundity type in fishes: Refining terms to reflect underlying processes and drivers

AbstractOnly with new technology and the ability to simulate systems are we beginning to understand fish reproductive processes at the individual scale over time. Traditionally, oocyte recruitment and fecundity type in fishes have been based on a “snapshot” approach, that is a histological micrograph and/or oocyte size frequency distribution of a spawning capable female has been assumed to indicate oocyte recruitment processes over time. This has important implications as the asynchronous and group‐synchronous oocyte recruitment patterns are associated with indeterminate and determinate fecundity types, respectively, and thus affect estimates of reproductive potential and spawning biomass (if using egg production methods). However, a historic review of these terms revealed that they are not applied consistently, with the same oocyte recruitment pattern often assigned to both of these by different scientists. The intent of this study is to develop a conceptual model of oocyte recruitment processes over time, key drivers, and resulting “snapshots” to refine oocyte recruitment terms, definitions, and the methods needed to identify them.

Can mesophotic reefs replenish shallow reefs? Reduced coral reproductive performance casts a doubt.

Mesophotic coral ecosystems (i.e., deep coral reefs at 30-120m depth) appear to be thriving while many shallow reefs in the world are declining. Amid efforts to understand and manage their decline, it was suggested that mesophotic reefs might serve as natural refuges and a possible source of propagules for the shallow reefs. However, our knowledge of how reproductive performance of corals alters with depth is sparse. Here, we present a comprehensive study of the reproductive phenology, fecundity, and abundance of seven reef-building conspecific corals in shallow and mesophotic habitats. Significant differences were found in the synchrony and timing of gametogenesis and spawning between shallow and mesophotic coral populations. Thus, mesophotic populations exhibited delayed or protracted spawning events, which led to spawning of the mesophotic colonies in large proportions at times where the shallow ones had long been depleted of reproductive material. All species investigated demonstrated a substantial reduction in fecundity and/or oocyte sizes at mesophotic depths (40-60m). Two species (Seriatopora hystrix and Galaxea fascicularis) displayed a reduction in both fecundity and oocyte size at mesophotic depths. Turbinaria reniformis had only reduced fecundity and Acropora squarrosa and Acropora valida only reduced oocyte size. In Montipora verrucosa, reduced fecundity was found during one annual reproductive season while, in the following year, only reduced oocyte size was found. In contrast, reduced oocyte size in mesophotic populations of Acropora squarrosa was consistent along three studied years. One species, Acropora pharaonis, was found to be infertile at mesophotic depths along two studied years. This indicates that reproductive performance decreases with depth; and that although some species are capable of reproducing at mesophotic depths, their contribution to the replenishment of shallow reefs may be inconsequential. Reduced reproductive performance with depth, combined with the possible narrower tolerance to environmental factors, further suggests that mesophotic corals may in fact be more vulnerable than previously conceived. Furthermore, we posit that the observed temporal segregation in reproduction could lead to assortative mating, and this, in turn, may facilitate adaptive divergence across depth.

Hermaphroditism induction of Sub-Lethal Dose of Atrazine and Atrazine-Nitrate on the Egyptian Toad, <i>Sclerophrys Regularis</i>

In this study, atrazine has been shown to act as a potent endocrine disruptor in amphibians either alone or in combination with nitrate under stable laboratory conditions, causing altered gonadal development at the sub-lethal concentrations. A control group and three treatments were tested; atrazine, nitrate, atrazine- nitrate treatments with doses of 300 μg/L, 200 mg/L and their combination, respectively. Atrazine exhibited increase the presence of testicular oocytes TOs in most treated specimens; furthermore, the sizes of seminiferous tubules were decreased compared to control group. Conversely no intersex individuals were detected in the nitrate treatment alone. The combined effects of nitrate and atrazine were not additive or synergistic but were similar to the effects of atrazine in raising the percentage of intersex but with increasing the size of testicular oocyte compared to atrazine alone. On the other hand, the control group in the present study did not contain the rudimentary testicular oocytes and showed normal structures. These gonadal abnormalities may reduce reproductive success and could be a major factor behind amphibian declines. Also, these negative impacts may be a bioindicator alarming the ecosystem disrupting caused by the uncontrolled apply of these chemicals in agriculture.

Preovulatory follicle growth dynamics indicates daily spawning synchronicity in wild-caught Atlantic horse mackerel off Portugal

The Atlantic horse mackerel, Trachurus trachurus, is a commercially important species whose daily spawning dynamics are not yet fully understood. Existence of spawning synchronicity would facilitate application of the daily egg production method for estimating the spawning biomass of the southern stock of the species in the Northeast Atlantic. The intent of this work was to study preovulatory follicle (PREF) growth dynamics in wild Atlantic horse mackerel to determine the existence of daily spawning synchronicity in the population. The size and translucency of hydrated oocytes increased significantly between early morning and late afternoon; this pattern was the same for two study years. This morpho-developmental shift led to the appearance of previously indistinguishable oil drops in ovarian whole mounts which is an easily observed microscopic descriptor of PREF growth. The results clearly suggested that PREF growth was well synchronized between Atlantic horse mackerel that were capable of spawning and provided evidence for the existence of daily spawning synchronicity in the population. Apart from sampling time of day, the size of hydrated oocytes was shown to be affected by body size: a 10 cm difference in maternal length corresponded to a 0.030 mm difference in the diameter of hydrated oocytes. In contrast, the size of early germinal vesicle migration oocytes was correlated neither to sampling time nor to maternal size which suggests that the abrupt increase in PREF size only commences after the onset of the actively spawning sub-phase.

Reproductive characteristics of three small-bodied cuttlefish in subtropical waters

Life history information for many commercially important cuttlefish is lacking, and thus the effects of long-term harvest remain unknown. The present study investigated the reproductive mode and seasonality of three small-bodied cuttlefish (Sepia opipara, S. plangon and S. rozella) in subtropical eastern Australia. Monthly samples were collected over 2 years and examined for gonadal development, size at sexual maturity, potential fecundity, size of mature oocytes and spermatophores, and temporal variation in maturity stages and gonadal weight relative to seawater temperature. All species showed reproductive characteristics consistent with a terminal multiple spawning mode, with S. opipara and S. rozella likely to be intermittent batch spawners, whereas S. plangon spawned continuously. Spawning occurred year round for all species, but S. opipara and S. rozella showed greater seasonality than S. plangon, peaking in autumn–spring. The biggest species, S. opipara, matured at a larger size than the others, had the greatest potential fecundity (the highest documented for any cuttlefish) and the largest spermatophore sizes. In contrast, S. rozella and S. plangon had much larger mature oocytes than S. opipara. These results further highlight the diversity of reproductive characteristics in cuttlefish, and the need for species-specific information for fishery and conservation management plans.

Reproductive biology and energetics of the brooding sea star Anasterias antarctica (Echinodermata: Asteroidea) in the Beagle Channel, Tierra del Fuego, Argentina

The brooding sea star Anasterias antarctica is distributed from the coast of Patagonia to the northern Peninsula of Antarctica. In the Beagle Channel, the females of A. antarctica brood their eggs for seven months and do not feed during this period. The endoparasite Dendrogaster argentinensis (Crustacea: Ascothoracica) causes castration in several species of Anasterias. We randomly collected four samplings of adults in May, August and October (brooding period) and January (non-brooding period). The gonad (GI) and pyloric caeca index (PCI) were calculated as organ wet weight (g) x 100/total wet weight (g). Each individual was sexed by microscopic examination of the gonads. Sex ratio, brooding females/non-brooding females and mature females/non-mature females ratios was 1:1. The male GI reached maximum values in January, when most individuals were sexually mature. The GI of non-brooding females reached its maximum during October when it was significantly higher than those from brooding females. The PCI was minimum in October, being lower in brooding females (August and October). During the non-brooding period, mature females had a significantly higher GI than non-mature females. The PCI did not vary neither between males, nor between mature and non mature females. By the end of the brooding period, non-brooding females showed a higher GI than the brooding females. This is explained by proliferation and increase of the oocytes size of non-brooding females. Mature females showed an incremented GI with presence of mature oocytes, while non-mature females exhibited more abundance of previtelogenic oocytes. Males showed synchronicity in reproductive condition. The females that have not brooded presented a process of active gametogenesis, reaching the summer with a high GI, therefore becoming mature females. Females that had brooded were probably lacking energy for new gonadal maturation. The pyloric caeca would be performing the role of a reserve organ in the brooding females, decreasing its size during the brooding period. Prevalence of D. argentinensis in A. antarctica was 11.06%. As this parasite was recorded in sea stars lacking gonads, these infected hosts could have been castrated. Rev. Biol. Trop. 65(Suppl. 1): S221-S232. Epub 2017 November 01.

Mesophotic coral-reef environments depress the reproduction of the coral Paramontastraea peresi in the Red Sea

With more than 450 studied species, coral reproduction is a well-known research field. However, the vast majority of coral reproduction research has focused exclusively on shallow reefs. The incentive for the present study was: (1) the recent accelerated global degradation of coral reefs; (2) the growing interest in mesophotic coral ecosystems (MCEs; 30–120 m depth) and their potential to serve as a larval source for shallow reefs; and (3) the lack of information on MCE coral reproduction. Here, we compare the reproduction and ecology of the depth-generalist coral Paramontastraea peresi between shallow (5–10 m) and mesophotic (40–45 m) habitats in the Gulf of Eilat/Aqaba, Red Sea. Field surveys were conducted to assess the living cover, abundance, and size frequency distribution of P. peresi. Four to six colonies from each habitat were sampled monthly between April 2015 and January 2017, and the gametogenesis cycles, fecundity, and oocyte sizes were measured. The reproductive cycle in the MCEs was shorter than in the shallow reef. Despite having larger polyps, the mesophotic colonies contained significantly smaller and fewer oocytes per polyp. In spite of the relatively stable environmental conditions of the MCEs, which may contribute to coral survival, scarcity of sunlight is probably a major energetic impediment to investment in reproduction by P. peresi at mesophotic depths. Further intensive reproductive studies in mesophotic reefs are thus required to assess the ability of corals in this environment to reproduce and constitute a larval source for depleted shallow-water reefs.

Reproductive strategy of Brachidontes rodriguezii (Bivalvia: Mytiloida) in the South-western Atlantic Ocean

This study consisted in the characterization of reproductive aspects such as oocytes morphology, temporal variation in size and frequency of oocytes and the reproductive cycle of the mussel Brachidontes rodriguezii. This species dominates the rocky intertidal coast of Buenos Aires province, Argentine and is a prey item of several exploited species; however, due to the introduction of piers and seawalls, as a consequence of anthropogenic activities, the species also inhabits sandy coasts. Mussels were collected from May 2011 to May 2013. Gonad tissue samples were processed according to standard histological procedures. Four gonadal development stages were described for males and females. According to the significant decrease in the frequency of mature oocytes, the spawning events were detected between February and May. However, some evacuation events were recorded in June–July. This study reports the first record of two modalities of hermaphrodites in this species; individuals with male and female gonads and individuals with both types of gametes within a gonad.

Role of sex steroids in gonadal differentiation of the mussel Choromytilus chorus (Bivalvia Mytilidae) (Molina 1782)

In Chile, the species Choromytilus chorus stands out for its ecological and economic importance. This mussel has a very particular condition in the colour of the gonad, being cream-yellow colour the male gonad and dark brown the female which is hardly desired by the consumers. In this context, the aim of this research is to determine the role that sex steroids have in gonadal differentiation of the mussel C. chorus. For this, juveniles with sizes 15–22 mm were selected, which were subjected to acute exposure in the form of bath for 60 days to four treatments: T1: dihydrotestosterone (DHT); T2: 17β-estradiol (E2), T3: fadrozole (F) and T4: DHT-F (DF), plus a control without chemicals. Each treatment as well as control included three replicates with 90 individuals each. The mussels undergoing treatment with E2 had a sex ratio (male:female) of 0.47 compared to 1:1 in the control group. In contrast, in the groups treated with DHT, F and DF the sex ratio changed to 2.0, 1.60 and 1.70 respectively. In the fertilization trials, all the mussels produced functional gametes, as they were able to generate gametes that were developed to morula, and veliger larva. Nevertheless, it should be noted that the size of oocytes from the group exposed to estradiol (61.12 ± 2.40 μm) was significantly higher than in other groups. These results support the hypothesis that sex steroids would be involved in sexual differentiation of marine bivalves.

Role of the venus kinase receptor in the female reproductive physiology of the desert locust, Schistocerca gregaria

Venus kinase receptors (VKR) are a subfamily of invertebrate receptor tyrosine kinases, which have only recently been discovered. They contain an intracellular tyrosine kinase domain and an extracellular Venus FlyTrap domain. VKRs have been functionally and pharmacologically characterized in only two invertebrate species, namely the human parasite Schistosoma mansoni and the mosquito Aedes aegypti, where they play a crucial role in oogenesis. Here, we report the characterization of a VKR in the desert locust, Schistocerca gregaria. We performed an in-depth profiling study of the SgVKR transcript levels in different tissues throughout the female adult stage. Using the RNA interference technique, the possible role of SgVKR was investigated. SgVKR knockdown had significant effects on ovarian ecdysteroid levels and on the size of oocytes during the vitellogenic stage. SgVKR is probably involved in the complex cross-talk between several important pathways regulating female reproductive physiology. Contrary to A. aegypti and S. mansoni, we cannot conclude that this receptor is essential for reproduction, since silencing SgVKR did not affect fecundity or fertility. Considering the evolutionary distance between A. aegypti and S. gregaria, as well as the differences in regulation of their female reproductive physiology, this article constitutes a valuable asset in better understanding VKRs.

Gigantic oocytes in the deep sea black coral Dendrobathypathes grandis (Antipatharia) from the Mar del Plata submarine canyon area (southwestern Atlantic)

The black coral Dendrobathypathes grandis (Cnidaria: Antipatharia) is studied for the first time in the southwestern Atlantic off Argentina. This is the only antipatharian reported from the Atlantic between 35°S and 54°S. Eleven specimens were collected at depths of 819–2204m during three expeditions to the Mar del Plata submarine canyon (2012–2013); seven were females. The species is gonochoric, and the polyps in female colonies contain up to nine oocytes per polyp, which can reach 1500µm in diameter. In contrast, the largest oocyte currently reported for antipatharians is 500µm, and usual diameters do not exceed 200µm. These large oocytes have over 20 times more volume than the biggest oocyte reported, but over 800 times more volume when compared with the common oocyte size of the group. Sperm size and morphology is similar to previous data from other species. As in previous studies, neither embryos nor larvae were found in any specimens. This species was previously only reported from waters off South Georgia Island, and so these specimens expand the known distribution north by 1800km.

Spawning area and season of butterfly kingfish (Gasterochisma melampus), a large scombrid adapted to cooler temperate southern water

In the present study we investigated spawning of the butterfly kingfish (Gasterochisma melampus), a Scombridae species distributed in circumpolar temperate waters of the Southern Hemisphere in the Atlantic, Indian and Pacific oceans. Using data from 25564 individuals collected by longline operations from 1987 to 1996, analysis of the gonadosomatic index, maturity based on oocyte size and the presence of hydrated eggs revealed that the spawning area was between longitude 85 and 130°W and latitude 28 and 41°S in the south-east Pacific Ocean, and that the spawning season was from mid-April to mid-July. Length–frequency data suggested that larger fish arrived and spawned earlier, whereas smaller fish did so later. The species has distinctive reproductive characteristics compared with other Scombridae: it produces large hydrated eggs 1.6mm in diameter, sea surface temperatures in the spawning area were as low as 14–18°C and more than 80% of fish were female. The south-east Pacific Ocean may be the only (and is at least the major) spawning area of the species. Butterfly kingfish is a single stock that migrates to the Atlantic, Indian and Pacific oceans to feed and returns to the south-east Pacific Ocean to spawn.

Temporal variations of potential fecundity of southern blue whiting (Micromesistius australis australis) in the Southeast Pacific

Fecundity is a key aspect of fish species reproductive biology because it relates directly to total egg production. Yet, despite such importance, fecundity estimates are lacking or scarce for several fish species. The gravimetric method is the most-used one to estimate fecundity by essentially scaling up the oocyte density to the ovary weight. It is a relatively simple and precise technique, but also time consuming because it requires counting all oocytes in an ovary subsample. The auto-diametric method, on the other hand, is relatively new for estimating fecundity, representing a rapid alternative, because it requires only an estimation of mean oocyte density from mean oocyte diameter. Using the extensive database available from commercial fishery and design surveys for southern blue whiting Micromesistius australis australis in the Southeast Pacific, we compared estimates of fecundity using both gravimetric and auto-diametric methods. Temporal variations in potential fecundity from the auto-diametric method were evaluated using generalised linear models considering predictors from maternal characteristics such as female size, condition factor, oocyte size, and gonadosomatic index. A global and time-invariant auto-diametric equation was evaluated using a simulation procedure based on non-parametric bootstrap. Results indicated there were not significant differences regarding fecundity estimates between the gravimetric and auto-diametric method (p > 0.05). Simulation showed the application of a global equation is unbiased and sufficiently precise to estimate time-invariant fecundity of this species. Temporal variations on fecundity were explained by maternal characteristic, revealing signals of fecundity down-regulation. We discuss how oocyte size and nutritional condition (measured as condition factor) are one of the important factors determining fecundity. We highlighted also the relevance of choosing the appropriate sampling period to conduct maturity studies and ensure precise estimates of fecundity of this species.

Novel functions for the RNA-binding protein ETR-1 in Caenorhabditis elegans reproduction and engulfment of germline apoptotic cell corpses

RNA-binding proteins (RBPs) are essential regulators of gene expression that act through a variety of mechanisms to ensure the proper post-transcriptional regulation of their target RNAs. RBPs in multiple species have been identified as playing crucial roles during development and as having important functions in various adult organ systems, including the heart, nervous, muscle, and reproductive systems. ETR-1, a highly conserved ELAV-Type RNA-binding protein belonging to the CELF/Bruno protein family, has been previously reported to be involved in C. elegans muscle development. Animals depleted of ETR-1 have been previously characterized as arresting at the two-fold stage of embryogenesis. In this study, we show that ETR-1 is expressed in the hermaphrodite somatic gonad and germ line, and that reduction of ETR-1 via RNA interference (RNAi) results in reduced hermaphrodite fecundity. Detailed characterization of this fertility defect indicates that ETR-1 is required in both the somatic tissue and the germ line to ensure wild-type reproductive levels. Additionally, the ability of ETR-1 depletion to suppress the published WEE-1.3-depletion infertility phenotype is dependent on ETR-1 being reduced in the soma. Within the germline of etr-1(RNAi) hermaphrodite animals, we observe a decrease in average oocyte size and an increase in the number of germline apoptotic cell corpses as evident by an increased number of CED-1::GFP and acridine orange positive apoptotic germ cells. Transmission Electron Microscopy (TEM) studies confirm the significant increase in apoptotic cells in ETR-1-depleted animals, and reveal a failure of the somatic gonadal sheath cells to properly engulf dying germ cells in etr-1(RNAi) animals. Through investigation of an established engulfment pathway in C. elegans, we demonstrate that co-depletion of CED-1 and ETR-1 suppresses both the reduced fecundity and the increase in the number of apoptotic cell corpses observed in etr-1(RNAi) animals. Combined, this data identifies a novel role for ETR-1 in hermaphrodite gametogenesis and in the process of engulfment of germline apoptotic cell corpses.

Reproduction of the cold-water coral Primnoella chilensis (Philippi, 1894)

This study examined the reproduction of a cold-water coral, Primnoella chilensis (Philippi, 1894) from the Comau and Reñihué fjords in Chilean Patagonia. Samples were collected in September and November of 2012 and April, June, and September of 2013 from three sites within the two fjords. The sexuality, reproductive mode, spermatocyst stage, oocyte size, and fecundity were determined using histological techniques. This species is gonochoristic with one aberrant hermaphrodite identified in this study. Reproduction was found to be seasonal, with the initiation of oogenesis in September and suggested a broadcast spawning event between June and September. The maximum oocyte size was 752.96µm, suggesting a lecithotrophic larvae. The maximum fecundity was 36 oocytes per polyp. Male individuals were only found in April and June. In June, all four spermatocyst stages were present. This suggests that spermatogenesis requires less time than oogenesis in P. chilensis.

Annual gametogenesis and reproductive effort of the limpet Cellana grata (Gould, 1859) (Gastropoda: Nacellidae) in a rocky intertidal beach at Ulleungdo Island off the east coast of Korea

Widely distributed from the northern coast of Vietnam to the northern Japan, the limpet Cellana grata (Gould, 1859) occurs commonly on the south and east coasts of Korea. Despite their wide distribution range, few studies have investigated the annual gametogenesis and reproductive effort of C. grata. In an attempt to understand the reproductive physiology of the limpet, we investigated the annual gametogenesis and reproductive effort of C. grata from Ulleungdo Island off the east coast of Korea. Histology revealed that the gonial mitosis commenced in January, as the female exhibited small oogonia (10–40 μm) in the follicle. From March to June, the oocyte size increased dramatically, and fully mature eggs (110–170 μm in diameter) appeared in early summer. First spawning males and females were observed in July, as the surface seawater temperature (SST) reached 22.1°C. The spawning male and females could be observed until the end of December. Gonad somatic index (GSI), a ratio of gonad mass to the total tissue weight, of the male ranged from 0.6 (April) to 17.9 (July), while the female GSI varied from 1.0 (February) to 18.3 (July). GSI of male and female declined rapidly from July to August, suggesting that the major purse of the spawning at the study site was between July and August. Our study suggested that the commercial catch of C. grata during July and August must be suspended at Ulleungdo Island, in order to protect the spawning limpets, which enhances C. grata recruitment and the population.

Large Cytoplasm Is Linked to the Error-Prone Nature of Oocytes

Chromosome segregation during meiosis in oocytes is error prone. The uniquely large cytoplasmic size of oocytes, which provides support for embryogenesis after fertilization, might be a predisposing factor for meiotic errors. However, this hypothesis remains unproven. Here, we show that cytoplasmic size affects the functionality of the acentrosomal spindle. Artificially decreasing the cytoplasmic size in mouse oocytes allows the acentrosomal spindle poles to have a better-focused distribution of microtubule-organizing centers and to biorient chromosomes more efficiently, whereas enlargement of the cytoplasmic size has the opposite effects. Moreover, we found that the cytoplasmic size-dependent dilution of nuclear factors, including anaphase inhibitors that are preformed at the nuclear membrane, limits the spindle's capacity to prevent anaphase entry with misaligned chromosomes. The present study defines a large cytoplasmic volume as a cell-intrinsic feature linked to the error-prone nature of oocytes. This may represent a trade-off between meiotic fidelity and post-fertilization developmental competence.