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The organisation of the ovarian interstitial tissue in the southern hairy-nosed wombat Lasiorhinus latifrons was investigated. Unlike in most other marsupials, the outer cortical region of the ovary contains abundant luteinised interstitial tissue that largely occurs in discrete lobules, many of which contain a localised area of non-cellular, highly eosinophilic and periodic acid-Schiff-positive material. The findings suggest that the latter arises from the zona pellucida that surrounded the oocyte in growing follicles and that the luteinised interstitial tissue thus developed from transformed theca interna of degenerated atretic follicles. It is hypothesised that this tissue synthesises and secretes progestogens, which may result in the long, and variable, oestrous cycle length that has been found to occur in this species.

Artibeus lituratus is one of the most recognized bat species in the neotropics, probably due to its high abundance in urban areas. Despite its wide distribution, the morphology and physiology of its reproductive organs remain unclear. Thus, the aim of the present study was to describe the morphology and histology of the female reproductive organs of this species, and evaluate morphological variations during its different reproductive phases (nonreproductive, pregnant, lactating, etc.). Thirty adult females were collected, divided into six (6) sample groups, according to the reproductive status, and submitted to anatomical and histological analyses. We show that the ovaries of A. lituratus are polarized, present a low number of interstitial glands, have large and well-developed Graafian follicles and a persistent corpus luteum, and may present a functional everted corpus luteum. Ovulation is simple, unilateral and nonpreferential, possibly alternated in successive ovulations, and may be sequential, with the occurrence of postpartum estrus. The uterus is simplex, the implantation is fundic and interstitial and the placenta is chorioallantoic, discoidal, and hemochorial. The female reproductive organs of A. lituratus share similarities with the pattern observed in human, which indicates that A. lituratus may be an interesting animal model in reproductive studies., (© 2019 Wiley Periodicals, Inc.)

Background: To investigate the therapeutic effects of menstrual blood derived mesenchymal stem cells (MB-MSCs) combined with Bushen Tiaochong recipe (BSTCR) on epirubicin induced premature ovarian failure (POF) in mice., Methods: Twenty-four female C57BL/6 mice of 6-8 weeks were intraperitoneally injected with epirubicin to induce POF, and then they were randomized into 4 groups of 6 mice each and treated with PBS, MB-MSCs, BSTCR, and MB-MSCs combined with BSTCR, respectively. Six mice of the same age were used as controls. Vaginal smear, TUNEL and hematoxylin-eosin staining were to observe estrous cycles, ovarian cell apoptosis and follicles. Enzyme-linked immunosorbent analysis determined serum estradiol, follicle-stimulating hormone (FSH) and anti-Müllerian hormone (AMH) levels. RT-qPCR and Western Blot analysis were to determine GADD45b, CyclinB1, CDC2 and pCDC2 expressions., Results: Epirubicin treatment resulted in a decrease in the number of primordial, primary, secondary and antral follicles, an increase in the number of atretic follicles and ovarian cell apoptosis, a decrease in estradiol and AMH levels, an increase in FSH levels, and estrous cycle arrest. However, MB-MSCs combined with BSTCR rescued epirubicin induced POF through down-regulating GADD45b and pCDC2 expressions, and up-regulating CyclinB1 and CDC2 expressions. The combined treatment showed better therapeutic efficacy than BSTCR or MB-MSCs alone., Conclusions: MB-MSCs combined with BSTCR improved the ovarian function of epirubicin induced POF mice, which might be related to the inhibition of GADD45b expression and the promotion of CyclinB1 and CDC2 expressions. The combined treatment had better therapeutic efficacy than BSTCR or MB-MSCs alone.

Previous studies have suggested that the smooth-billed ani (Crotophaga ani, Linnaeus, 1758) breeds opportunistically following unpredictable rainfall in drought areas. To obtain proof of this phenomenon, the present study described and compared reproductive morphology and cell proliferation in the gonads of free-living smooth-billed anis during a wet season (April to June 2012) and the following dry season (July to September 2012) in a semiarid area using light and electron microscopy (transmission and scanning) and the AgNOR method. The morphological findings indicated distinct levels of reproductive activity related to seasonal changes. Morphological and morphometric analyses of the gonads confirmed intense gametogenic activity during the wet season, whereas gonadal involution occurred after rainfall ceased. The sizes of the testes and ovaries were significantly reduced compared to those in the wet season. The volumetric fraction of the seminiferous tubules in the testis decreased considerably, and no preovulatory follicles were detected in the ovary in the dry season. Moreover, the AgNOR count in the gonads revealed a significant decline in cell recruitment for gametogenesis after rainfall ceased. The histological findings indicated partial gonadal activation throughout the dry season. The analysis of the seminiferous epithelium confirmed the early testicular recrudescence phase, and sporadic postovulatory follicles indicated random ovulation during this time. The excurrent ducts and the oviduct also underwent remarkable involution in the dry season. Taken together, these findings confirm opportunistic breeding by smooth-billed anis in a semiarid habitat and suggest that gonadal recrudescence has been established as a reproductive strategy to cope with unexpected precipitation events., (© 2019 Wiley Periodicals, Inc.)

The ovary is a structurally dynamic organ that alters with age. Modifications in the paracrine status influence the capacity of aging oocytes to develop normal embryos. Despite the importance of understanding the cellular and molecular mechanism involved in the process of ovarian aging, histological changes remain poorly understood. Correlating the process of folliculogenesis and somatic cell function during ovarian aging is essential to explain the reproductive decline of aged mammalian species, including humans. Here, we performed a morphological and immunohistological study on the ovaries of chinchilla rabbits that varied in age from one to 34-months. The spatiotemporal expression of the cholesterol side-chain cleavage cytochrome P450scc (CYP11A) and the smooth muscle actin (SMA) were analyzed. A significant histological rearrangement of immunodetected cells in theca interna, theca externa and the interstitial tissue around the follicles occurred. The expression of CYP11A1 decreased considerably in antral follicles of aging ovaries. Moreover, we found that the secondary interstitial gland developed extensively, and a remarkable rearrangement of the surface epithelium occurred in aging ovaries. In contrast to ovaries during the reproductive period, the immunohistological changes demonstrate that the interstitial gland became the most abundant tissue during the aging of ovaries. Thus, the current study provides new data for understanding the alteration of somatic cell function in elderly ovaries and how this affects their declined fertility.

Laboratory and synchrotron X-ray tomography are powerful tools for non-invasive studies of biological samples at micrometric resolution. In particular, the development of phase contrast imaging is enabling the visualization of sample details with a small range of attenuation coefficients, thus allowing in-depth analyses of anatomical and histological structures. Reproductive medicine is starting to profit from these techniques, mainly applied to animal models. This study reports the first imaging of human ovarian tissue where the samples consisted of surgically obtained millimetre fragments, properly fixed, stained with osmium tetroxide and included in epoxydic resin. Samples were imaged by the use of propagation phase contrast synchrotron radiation micro-computed tomography (microCT), obtained at the SYRMEP beamline of Elettra light source (Trieste, Italy), and X-ray absorption microCT at the Theoretical Biology MicroCT Imaging Laboratory in Vienna, Austria. The reconstructed microCT images were compared with the soft X-ray absorption and phase contrast images acquired at the TwinMic beamline of Elettra in order to help with the identification of structures. The resulting images allow the regions of the cortex and medulla of the ovary to be distinguished, identifying early-stage follicles and visualizing the distribution of blood vessels. The study opens to further application of micro-resolved 3D imaging to improve the understanding of human ovary's structure and support diagnostics as well as advances in reproductive technologies.

We evaluated the effect of photoperiod on ovarian morphology, reproductive hormone secretion, and hormone receptor mRNA expression in layer ducks during the pullet phase. A total of 480 71-d-old Jinding layer ducks were randomly divided into 5 groups that received 6L (hours of light):18D (hours of darkness), 8L:16D, 10L:14D, 12L:12D, or 14L:10D, respectively. Each group had 6 replicates with 16 birds each. The photoperiod feeding trial lasted 80 d until 150 d of age. The age at first egg (AFE), the total number, and weight of eggs increased linearly with increasing photoperiods (P < 0.05); lower values of AFE occurred with photoperiods ≥8 h, whereas a higher total number and weight of eggs occurred with photoperiods ≥10 h, compared with 6L:18D (P > 0.05). Oviduct weight, ovary percentage, and initial and bare stroma (weight and percentage) increased quadratically with increasing photoperiods (P < 0.05), and 10.24, 10.01, and 10.10 h were the optimal photoperiods for oviduct weight, bare stroma (follicles ≥2 mm in diameter removed) weight, and bare stroma percentage, respectively, as calculated from reliable regression equations (R2 ≥ 0.5791). Compared with 6L:18D, 10L:14D had a higher total large white follicle weight, small yellow follicle number, and weight (P < 0.05). In addition, higher serum levels of follicle-stimulating hormone, luteinizing hormone, and progesterone were observed with ≥10-h photoperiods (P < 0.05), as were levels of hormone receptor mRNA expression in ovarian follicles (P < 0.05), with the highest values for both measures at 10L:14D. In the hypothalamus, mRNA expression of gonadotropin-releasing hormone increased in ≥8-h photoperiods, with the highest value at 10L:14D. In contrast, gonadotropin-inhibitory hormone increased in photoperiods ≥12 h (P < 0.05). In conclusion, an appropriate photoperiod led to early sexual maturity and improved the development of reproductive organs and ovarian follicles through effects on reproductive hormones and their receptors; 10 to 10.24 h is an adequate photoperiod for layer ducks during the pullet phase., (© 2019 Poultry Science Association Inc.)

Equine Chorionic Gonadotropin (eCG) previously known as Pregnant Mare Serum Gonadotropin (PMSG) has been used for decades in regulating reproduction in various domestic animal species. Its use necessitates a good measurement of its bioactivity in commercial preparations. The EUROPEAN PHARMACOPEIA (EP 7.0) recommends 5-6 subcutaneous injections in immature female rats for the in vivo bioassay for eCG as in the case for measurement of FSH bioactivity in the Steelman & Pohley assay (1953). This recommendation is in marked contrast with the classical and long-time used PMSG assay of Cole & Erway (1941) that includes only one subcutaneous injection, 3 days before measurement of ovarian weight. As this difference introduces much confusion in the determination of eCG/PMSG bioactivity in commercial sources, we have performed parallel assays of several PMSG preparations, with both protocols. The single-injection protocol takes into account the long half-life of eCG in bloodstream and provokes an ovarian stimulation at lower concentrations than the multiple-injection protocol. As the single-injection protocol also mimicks the protocol used in cattle, it is preferable to the multiple-injection protocol of the current EP., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Heterandria formosa is a viviparous poeciliid native of the southeastern of United States of America. H. formosa exhibits unique reproductive features as: (a) production of extremely small eggs with scarce quantity of yolk (microlecithal eggs), (b) consequently, a high level of matrotrophy and development of a complex follicular placenta, (c) ovarian sperm storage that allows the continuous fertilization of oocytes and production of offspring and (d) development of high degree of superfetation. The degree of superfetation refers to the number of broods in different simultaneous stages of gestation. Morphological evidence of the degree of superfetation in H. formosa has not been documented. Therefore, and because of the general interest in the complex process of superfetation, the goal of this study is to morphologically define the degree of superfetation of H. formosa through two procedures: (a) histological analysis of entire ovaries in gestation and (b) dissection of visible embryos and the histological analysis of the remaining ovarian tissue. Results indicate that H. formosa can gestate up to seven broods at the same time., (© 2019 Wiley Periodicals, Inc.)

Techniques for counting ovariole number in virgin and mated Apis mellifera L. queens have been described in previous studies. Having a systematic and fast way to collect this measurement can help accelerate bee breeding programs, because selection decisions can be taken faster. The aim of this work was to develop an efficient histological method to preserve ovaries that allows assessing the number of ovarioles in newly emerged virgin queens, and also in mated queens, in a shorter time than the methods already published. The proposed method resulted in images suitable for ovariole counting in both newly emerged and mated queens, and the total histological process took less than 10 h. This method provides the optimization of the histological procedure for research breeding programs that use ovariole number as selection criteria for improving reproduction and production traits., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America.)

Cellular aspects of oocyte development of the Mexican rivulus Millerichthys robustus were morphologically described in order to analyze ovarian function and the cellular recruitment dynamics associating it with life history strategies of annual killifishes. Millerichthys is an iteroparous batch spawner with continuous oocyte recruitment and indeterminate fecundity with asynchronous development of the follicles. It has two ovaries of cystovarian type, with a central lumen, which communicates with the outside through the caudal region of the ovary, that is, the gonoduct. From the walls of the ovary, irregular lamellae composed of germinal epithelium and vascularized stroma project. Oogenesis starts with oogonial proliferation, found alone or in nests within the germinal epithelium. The oogonia come into meiosis becoming oocytes and advancing to the chromatin nucleolus stage and to early primary growth stage. Folliculogenesis is completed in the primary growth stage and cortical alveoli step. Follicles moves toward the stroma, but they continue to be attached to the germinal epithelium through the basement membrane until ovulation. The inclusion of fluid yolk in the follicles during the secondary growth stage was observed. During ovulation, the follicle collapsed, the oocyte was released into the lumen, and the constitutive elements of the post-ovulatory follicle complex remained in the stroma., (© 2019 Wiley Periodicals, Inc.)

In this paper, we describe the morphology of the female Bagrada hilaris (Burmeister) reproductive system and develop a physiological age-grading system related to egg production. The female reproductive system is composed of two meroistic and telotrophic ovaries each containing 5-6 tubular ovarioles. The ovarioles unite into the lateral oviduct which combine to form the common oviduct. The ovarioles are composed of two regions; the distal germarium and the tubular vitellarium which contains maturing follicles. Each follicle is surrounded by a layer of cells; the follicular epithelium. As the follicle passes from the ovariole to the lateral oviducts the follicular epithelial cells slough off and accumulate in the base of the ovarioles and are known as follicular relics. The continuum of ovarian development is divided into two categories: nulliparous ('nonreproducing') and parous ('reproducing'). The nulliparous category is characterized by the absence of follicular relics or eggs in the oviducts as opposed to the parous category where follicular relics and, in many cases, eggs occurred in the oviducts. The nulliparous category is divided further into two stages; N1 and N2 based on ovariole differentiation. The parous category is divided into three stages; P1, P2, and P3, based on the quantity and appearance of follicular relics. Females characterized as P3 produced three times more eggs (79.2 eggs ± 5.7) than females characterized as P1 (27.1 eggs ± 6.0)., (Published by Oxford University Press on behalf of Entomological Society of America 2019. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

Background: This study determined the anatomical and histological structure of the salpinx and ovary of the Anatolian wild goat (C. aegagrus aegagrus)., Materials and Methods: The salpinges and ovaries from four wild goats of similar ages were used. These goats were brought to the Kafkas University Wildlife Rescue and Rehabilitation Centre (Kars, Turkey) for various reasons such as traffic accidents or firearm injury, but either could not be saved or were euthanased., Results: The mean length of the ovary was 13.22 ± 1.27 mm, width was 8.46 ± 0.88 mm, thickness was 5.67 ± 0.79 mm, and weight was 0.59 ± 0.17 g. The mean length of the salpinx was 58.11 ± 14.02 mm, width was 0.80 ± 0.22 mm, thickness was 0.41 ± 0.01 mm, and weight was 0.30 ± 0.08 g. The salpinx consisted of three different regions (infundibulum, ampulla, and isthmus). These regions consisted of tunica mucosa, tunica muscularis, and tunica serosa. Prismatic epithelial cells were observed in the lamina epithelialis of the tunica mucosa in every region., Conclusions: The data resulting from this study regarding anatomical and histological structures of the salpinx and ovary in the genital system organs of Anatolian wild goat will support other studies seeking to improve reproductive efficiency and eliminate the danger of extinction of this species.

Even though tardigrades have been known since 1772, their phylogenetic position is still controversial. Tardigrades are regarded as either the sister group of arthropods, onychophorans, or onychophorans plus arthropods. Furthermore, the knowledge about their gametogenesis, especially oogenesis, is still poor and needs further analysis. The process of oogenesis has been studied solely for several eutardigradan species. Moreover, the spatial organization of the female germ-line clusters has been described for three species only. Meroistic ovaries characterize all analyzed species. In species of the Parachela, one cell per germ-cell cluster differentiates into the oocyte, while the remaining cells become the trophocytes. In Apochela several cells in the cluster differentiate into oocytes. Vitellogenesis is of a mixed type. The eggs are covered with the egg capsule that is composed of two shells: the thin vitelline envelope that adheres to the oolemma and the thick three-layered chorion. Chorion is formed as a first followed by vitelline envelope. Several features related to the oogenesis and structure of the ovary confirm the hypothesis that tardigrades are the sister group rather for arthropods than for onychophorans.

Mytilus galloprovincialis female specimens were collected from two mussel farms located in two sites next to Castel dell'Ovo, a historical complex located in the Naples Bay. Such sites were named, respectively, A-area and B-area for the different microbiological parameters so that mussels from A-area can be sold without purification, whereas mussels from B-area must be purified before sale. The mussels were collected during the nonreproductive (summer 2009) and reproductive periods (autumn 2009). Gonadosomatic index, structural organization of the ovary, presence of apoptosis, estrogen receptors expression, as well as the bisphenol A (BPA) content in the ovaries, were evaluated. Ovaries from specimens collected in area B showed a different and significant distribution of the investigated biomarkers as well as of BPA content in respect to those measured in the A-area specimens, confirming that mussels are valid sentinel organisms to biomonitor in the Naples bay too., (© 2018 Wiley Periodicals, Inc.)

The subphylum Chelicerata represents one of the oldest groups among arthropods and comprises more than a dozen orders. Representatives of particular orders differ significantly in their external morphology, reproductive biology, behavior, and structure of internal organs, e.g. of the respiratory system. However, in almost all chelicerates (excluding some mites) the female gonads show a similar architecture. In this chapter, the chelicerate-type ovary structure and the course of oogenesis are described. Structural and functional diversities of the chelicerate-type ovary in non-matrotrophic and matrotrophic arachnids are also presented.

Bisphenol A (BPA), an ubiquitous environmental endocrine disruptor chemical, disturbs the mRNA expressions of steroidogenic genes and subsequently steroid hormone synthesis in mammals and aquatic species. However, the underlying regulation mechanisms are barely understood, especially in fish. To explore the regulation mechanism, we exposed female rare minnow Gobiocypris rarus (G. rarus) to BPA at a nominal concentration of 15 μg/L for 7 and 14 days in the present study. Results showed significant increase of gonad somatic index (GSI) and serum estradiol (E2) levels in response to BPA at day 14. The 7-day BPA exposure notably repressed the expression of two ovarian steroidogenic genes (star and hsd11b2) and suppressed their capacity of estrogen response elements (ERE) to recruit estrogen receptor (ER), while the 14-day BPA treatment remarkably induced transcript of hsd3b and enhanced the capacity of ERE to recruitment ER in ovaries. Furthermore, the 7-day BPA exposure caused DNA hypermethylation of star (CpGs: -742 bp and -719 bp) and hsd11b2 (CpG: -1788 bp). However, 14-day BPA exposure resulted in DNA hypomethylation of hsd3b (CpG: -181 bp). Correlation analysis revealed that the DNA methylation levels at specific CpGs in star, hsd3b and hsd11b2 were significantly correlated to their mRNA levels and ER-EREs interactions. These findings suggest that the disturbed steroidogenesis and the transcripts of ovarian steroidogenic genes might attribute to the altered DNA methylation status of these ovarian steroidogenic genes in response to BPA., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: The effects of methanol extract of Parquetina nigrescens were studied on histomorphometry and protein expression (SDS-PAGE) from the ovaries and uteri of wistar rats., Methods: 30 sexually matured rats were used for the study with 10 each in the control and treatment 100 mgkg -1 and 400 mgkg -1 groups. The extract was orally administered for 14 days. Histological sections of tissues collected presented no abnormalities., Results: An increase in the number of developing and matured follicles were observed during the study in the treated groups compared to the control in the follicular and the luteal phases. The corpora lutea in the treated groups were fewer in number to that of the control in the follicular phase and in the luteal phase. Sections of the uterine horns showed significant narrowing in the lumen diameter and increases in epithelial height with increased laydown of the lamina propria in the treated groups. The expression of protein bands fractionated during the study, confirm the presence of proteins expressed repeatedly from the ovary and uterine horns in the follicular and luteal phases at the 70 kDa and 63 kDa regions., Conclusions: The study concluded that the methanol extract of the plant increased folliculogenesis on the ovary, secretory activity in the nuclei of the epithelium and the fibroplasia of the lamina propria while narrowing the lumen of the uterine horns which are similar to the effects of oestrogen or oestrogen-like substances on these reproductive organs and may have an effect on the abundance of protein expressed in the follicular phase., Competing Interests: The authors declare that they have no conflict of interest., (© Georg Thieme Verlag KG Stuttgart · New York.)

Knowledge of the reproductive cycle in exploited species is important for a sustainable management of fisheries. Standardized scales to assess maturity stages are a fundamental tool to understand the demographic composition of exploited populations. Staging scales for female Norway lobster, Nephrops norvegicus, have been subject to a series of changes, and multiple inconsistent scales are in use in different fisheries regions. A unified, evidence-based scale has not previously been established. We reviewed previous staging scales for the female ovary maturation and propose a revised scale based on the correlation between macroscopic and microscopic ovary characteristics. To provide better-informed tools for future stock assessment, female stages were characterized through external observation on ovary color and size, and the progress of vitellogenesis. This study clarifies several biological phases that were conflated in previous scales. First, we demonstrate how to distinguish between immature ovaries in juvenile females versus the earliest ovary maturation stage in adults. Second, the new scale differentiates between "mottled" ovaries seen in two separate biological stages: the spent ovaries that undergo partial resorption in berried females, versus ovaries of females which failed to spawn and undergo full resorption. To ensure consistent application, colors are assessed relative to international standards (RAL/Pantone). This new, practical staging scheme clarifies the correlation between microscopic characteristics and macroscopically observable details in ovary maturation. Adoption of this unified staging scale will improve maturity analyses, help to identify stocks with potentially reduced reproductive capacity, and facilitate broad-scale comparisons., (© 2018 Wiley Periodicals, Inc.)

Histological examination of ovarian morphology was conducted on the monkfish, Lophius americanus. The ovary is of the cystovarian type, that is, with a lumen. However, ovarian morphology is quite unique, compared to other fishes, in that ovarian lamellae only originate along the ventral ovarian wall rather than around the periphery. They may branch at their base, but are otherwise unbranching. Furthermore, the germinal epithelium that is, the site of germ cell production, is described for the first time in L. americanus. The germinal epithelium is uniquely restricted around the base of each lamellae rather than being distributed along lamellar epithelia. Folliculogenesis, the process whereby ovarian follicles are formed, is completed in the Perinucleolar Step of the Primary Growth Stage. As oocytes grow and mature, they are increasingly observed towards the apex of the unbranched lamellae where ovulation occurs. The appearance of small vesicles within yolk at the vegetal pole was observed in oocytes during maturation. Prior to ovulation, cells of the ovarian epithelium produce a mucogelatinous matrix that forms the veil in which eggs, after fertilization, become suspended in the water column until hatching., (Published 2018. This article is a U.S. Government work and is in the public domain in the USA.)

The processes of follicular development, ovulation, egg capture, and egg transport vary among vertebrates. Therefore, analysis of the reproductive tract of several lineages is needed for understanding the evolutionary changes of the reproductive system. In turtles, the ovulated eggs are released into the coelomic cavity and taken up by one of the two oviducts, a phenomenon called extrauterine migration of eggs. However, the process of egg uptake in lizards is different. The egg is ovulated directly into the infundibulum, and oocyte uptake by the contralateral oviduct rarely occurs. The same pattern has been hypothesized to occur in snakes. To test this hypothesis, we analyzed the reproductive tract of female Micrurus corallinus preserved in zoological collections. We examined the anatomical characteristics of the infundibulum and ovary and compared the reproductive output between the ovaries to verify the mechanism of egg capture and the anatomical viability of extrauterine migration of eggs. The reproductive output of the right ovary was higher than that of the left ovary, and the higher number of eggs in the right oviduct is due exclusively to the production of the ipsilateral ovary. Several anatomical features prevent extrauterine migration of eggs, including the asymmetry of the reproductive system, the arrangement of the ovarian follicles in a single row, and the formation of a wrapping around the ovary and infundibulum by the visceral pleuroperitoneum membrane (preventing against ectopic eggs). Therefore, the hypothesis of egg capture by the contralateral oviduct is anatomically infeasible in M. corallinus and possibly in other snakes. Anat Rec, 301:1936-1943, 2018. © 2018 Wiley Periodicals, Inc., (© 2018 Wiley Periodicals, Inc.)

The aim of this study was to evaluate two-dimensional and Doppler ultrasonographic changes of feline ovaries before and during puberty. Nine, 3-month-old female cats were followed until puberty (Day 1). Two-dimensional and Doppler ultrasound evaluations of the ovaries were carried out on Days -50, -20, -7 and 1. Longitudinal and transverse sections of the ovaries were measured and all anechoic spherical structures were considered to be follicles. The number of follicles >1 mm and the maximum diameter of the largest follicle were recorded. Peak systolic velocity and end diastolic velocity of intraovarian arteries were also measured to automatically calculate the resistive index (RI). The mean ovarian longitudinal diameter increased gradually throughout the study from 8.6 to 10.7 mm (p < 0.05). While four cats presented multiple anechoic spherical structures <1 mm diameter throughout the study, the remaining five animals had these structures only on Days -50 and -20. On Days -20, -7 and 1, the mean number of follicles were 1.4 ± 0.7, 2.5 ± 0.8 and 4.8 ± 1 respectively (p < 0.01). The largest follicles at the same time points were 1.1 ± 0.2 mm, 1.9 ± 0.3 mm and 2.6 ± 0.5 mm respectively (p < 0.05). The RI of the intraovarian arteries declined throughout the study period (p < 0.01). It is concluded that, in female cats, ovarian dimensions, follicle number and intraovarian blood flow increased from 3 months of age to puberty., (© 2018 Blackwell Verlag GmbH.)

Steroidogenic activity in the equine ovary from birth to puberty has been poorly investigated. This study aimed to examine the capability of the ovarian follicles of prepubertal and pubertal fillies to produce steroid hormones and to evaluate the expression and cellular localization of androgen receptor (AR) in their ovaries. The ovaries of 6-18 month-old fillies were divided into two groups: prepubertal (PrP) - without preovulatory follicle (pF) and corpus luteum (CL), and ovulating/postpubertal (Ov/pB) - with pF and/or CL in at least one of the gonads. Adult mares (Me) were used as a control. The concentration of progesterone (P4), testosterone (T) and estradiol (E2) in follicular fluid (FF) was measured by radioimmunoassay. AR distribution was assessed by immunohistochemistry, while AR protein expression was examined by Western blot analysis. In the female groups, E2 concentration in FF of small follicles (<10 mm) was low and increased with the diameter of the follicle reaching the greatest value in pF (Ov/pB and Me group). In follicles (11-30 mm) of PrP fillies, the concentration of E2 was similar to that from Ov/pB fillies, but less than half (P < 0.05) than in Me follicles. In FF from all classes of follicles of Ov/pB fillies, the concentration of all steroids was similar to that in Me. AR immunolocalization, predominantly nuclear, was observed in all types of follicular cells (granulosa and theca cells) as well as in stroma and luteal cells. The pattern of staining was dependent on the follicle size and the group of females. In smaller antral follicles and in pF, the nuclear AR staining in granulosa cells was stronger than that found in follicles of 21-25 mm. In theca interna cells of pF, both nuclear and faint cytoplasmic reactions were seen. In luteal cells, AR labeling was noted in the nuclei and the cytoplasm: the strongest one in the early CL and almost negative in the late CL. AR protein expression in filly and mare ovarian tissues was confirmed by Western blot analysis and detected as a single band at approximately 110 kDa. In summary, the ovaries of fillies aged at least 6 months are capable of active steroidogenesis. ARs are present either in the cell nuclei or cytoplasm of all compartments of the equine ovary. AR expression in follicular and stroma cells may indicate the sensitivity of the filly ovarian tissue to androgens, the impact of androgens on folliculogenesis and the development of the equine ovary via a receptor-mediated pathway., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Detection of ovarian follicle/cysts and monitoring its growth is vital in infertility treatment in women. Ultrasound imaging technique is used for recognition of ovarian follicles and cysts. Profound variation in location, shape, size and color is seen in follicles. Human interpretation of follicles gives a chance for misinterpretation and false diagnosis. Follicle recognition becomes a challenging task due to the non-homogenous nature of the follicles and presence of speckle noise. To overcome this problem, computer assisted recognition of ovarian follicle and cysts followed by ovarian classification were proposed. Discrete wavelet transform (dwt) was used for despeckling. Texture and intensity based segmentation methods were used for automatic recognition. Classification of ovary was done based on ovarian morphology. This novel method serves as a decision support system for the medical expert. The efficiency of the proposed texture and intensity based ovarian classification (TIOC) method was demonstrated using various performance indices like sensitivity, specificity, accuracy, precision, Mathew's correlation coefficient and receiver operating characteristic curve. The resultant images obtained from the TIOC method was compared with the control images and existing methods for validation.

The present study was aimed at evaluating the effect of male presence on ovarian maturation in juvenile females and the role of potential chemical, visual and tactile cues emitted by males in that physiological process. A highly gregarious caridean shrimp with sexual dimorphism, Neocaridina davidi, was used as experimental model. We tested the hypothesis that male presence accelerates ovarian maturation, mainly through chemical cues. Two experiments were performed. In Experiment 1, juvenile females were reared with adult males, adult females or alone, allowing full contact among shrimps. In Experiment 2, these treatments were evaluated allowing chemical and visual communication, only visual communication, or only chemical communication among shrimps. In both experiments juvenile females were observed once a week under a stereomicroscope to determine ovarian growth rate. Although male presence was not necessary for ovarian maturation, it clearly accelerated the rate of ovarian growth, particularly in the last maturation phase. This lead to relatively longer mature ovaries with higher lipid content. On the contrary, the presence of adult females delayed ovarian maturation in juvenile females, while females reared alone showed an intermediate ovarian growth. All these results suggest that adult males release certain cues that stimulate ovarian maturation, while adult females release cues that delay this physiological process. Neither visual cues nor chemical cues released at a distance from females were responsible, either alone or in combination, for the observed effects. Ovarian growth was only influenced when shrimps were allowed to interact freely, probably because of the "mounting" behavior of males towards females. Tactile cues and/or potential chemical cues released by males during this behavior may mediate male stimulatory effect on ovarian growth. Altogether, present results partially support our initial hypothesis and contribute to increase the limited amount of information available on the role of intraspecific multimodal communication in non-behavioral reproductive processes in invertebrate species., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The objective of this study was to assess the efficiency and clinical safety of postnatal administration of a GnRH agonist on canine puberty postponement. Sexual steroids and histological gonadal changes were also described. Twenty-four littermate puppies were randomly assigned to: Deslorelin acetate 18.8 mg sc (DESLO; n = 12) or Placebo: sc (PLACE; n = 12) postnatally. The dogs were clinically and endocrinologically followed up until puberty when they were gonadectomized and their gonads histomorphometrically studied. Deslorelin postponed the age of puberty (72.7 ± 4.8 vs. 35.8 ± 1.9 weeks; P < 0.01) in these dogs. At the time of this submission, 3 DESLO dogs (108 weeks old) remain non-pubertal. All dogs concluded growing at a similar age (29.75 ± 2.44 vs. 29.25 ± 0.90 weeks; P > 0.1) independently of their group and pubertal status. None of the females had side effects while the 2 non pubertal DESLO males presented bilateral cryptorchydism. All the bitches ovulated at puberty (P > 0.1) and the 2 DESLO that were mated became pregnant. Deslorelin postponed basal serum sexual steroids up to puberty in both genders (P < 0.01). The histomorphometrical study of the testes revealed that the tubular diameter (P < 0.05), germinal epithelium height and composition (P < 0.01) were decreased in DESLO group. Ovarian structures did not differ between treatments (P > 0.05). It was concluded that postnatal deslorelin decreased sexual steroids reversibly postponing puberty in both genders without side effects in bitches and causing 2/6 of cryptorchydism and impairment of testicular histomorphometry in male dogs., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Andean frogs of the genus Telmatobius occur at high elevations, they have an aquatic mode of life and large tadpoles. There are more than 60 species that closely resemble one another and have low values of genetic divergence. However, the skeleton, particularly the cranium, is interspecifically variable with respect to the different levels of development of some elements. Heterochrony is considered to have played a prominent role in generating phenotypic variation, especially among closely related species. Herein, the developmental origins of the adult cranial configuration of two species of Telmatobius are explored. The interactions among larval and postmetamorphic growth, sexual maturation, and ossification sequence in T. oxycephalus and T. rubigo are studied. Although there are no substantial changes in the sequences of ossification of the cranium, it is likely that differential timing of larval periods is related to adult cranial characters. The prolonged larval development of T. rubigo may result in peramorphic configurations of bones that ossify during pre-metamorphosis. This long developmental time would also explain why the gonads of T. rubigo are highly differentiated by the end of metamorphosis. In this species, sexual maturation may be attained precociously in relation to metamorphosis, thereby reducing postmetamorphic developmental time of late-onset bones, which have paedomorphic configurations (e.g., vomer, neopalatine, and columella). An inverse pattern characterizes T. oxycephalus, suggesting that the duration of larval life is related to skeletal configuration in Telmatobius., (© 2018 Wiley Periodicals, Inc.)

Teleosts possess unique features of the female reproductive system compared with the rest of vertebrates, features that define the characteristics of their viviparity. Viviparity involves new maternal-embryonic relationships detailing the most diverse structures during gestation that include embryonic nutrition. In order to analyze the morphological features of the complex nutrition in viviparous teleosts during intraovarian gestation, this study utilizes the goodeid Xenotoca eiseni as a model. Ovarian gestation in X. eiseni, as in all goodeids, is intraluminal; the early embryo moves from the follicle to the ovarian lumen where gestation continues. The scarce yolk in the oocytes implies that the initial lecithotrophy is replaced by matrotrophy, with nutrients provided via maternal tissues. The nutrients are absorbed by the embryo mainly by trophotaenia, extensions of the embryonic intestine into the ovarian lumen. This histological study analyses the structures involved in these two types of nutrition and when they occur during gestation in X. eiseni. The morphology displayed in this study demonstrated the extended simultaneity of lecithotrophy and matrotrophy during gestation with the progressive reduction of lecithotrophy and increase of matrotrophy. Similarly, it describes the development of complex embryonic structures for metabolic exchange with the maternal tissues associated with matrotrophy; specifically the branchial placenta and mainly the trophotaenia., (© 2018 Wiley Periodicals, Inc.)

Aim: Ligation of major vessels supplying ovaries may alter hormones and ovarian reserve due to disturbances of vascular circulation. Our purpose is to measure serum anti-Müllerian hormone (AMH) levels and ovarian volume in patients who had internal iliac artery ligation (IIAL) and/or IIAL plus hysterectomy due to uterine atony., Methods: Patients who underwent IIAL and IIAL+ hysterectomy were evaluated 6 months after their operations and were compared with the control group. The hormones, ovarian volume and antral follicle count (AFC) were measured in each group., Results: Serum AMH levels in the post-partum 6th month interval were lower in the IIAL group than in the control group and were the lowest in the IIAL+ hysterectomy group. Similar to AMH results, AFC and ovarian volumes were also lowest in the IIAL+ hysterectomy group., Conclusion: IIAL and hysterectomy are lifesaving interventions during peripartum hemorrhage; however, they might alter ovarian reserve in the short term., (© 2018 Japan Society of Obstetrics and Gynecology.)

Objectives: The current study aimed to investigate FSH receptor binding inhibitor (FRBI) effects in the expressions of FSH receptor (FSHR) and estrogen receptor-beta (ERβ) in the mice ovaries at the gene and protein levels, also to find the potential efficacy of FRBI on suppressing ovarian cancer through down-regulating over-expression of FSHR and ERβ in the normal ovarian tissues., Methods: 180 female mice were randomized into six groups (n = 30). Mice of FRBI-1, FRBI-2 and FRBI-3, FRBI-4 were intramuscularly injected with FRBI of 10, 20, 30 and 40 mg/kg, respectively, for five consecutive days. The qPCR and Western blotting were used to determine expression levels of FSHR and ERβ mRNAs and proteins in mouse ovaries., Results: The ovarian cortex thickness (OCT) of the FRBI-4 group were less than that FSH group on day 30 (P < 0.05). The numbers of secondary follicles (SF) and the maximum transverse diameters (MTD) of secondary follicles of FRBI-3 and FRBI-4 groups were decreased as compared to FSH group (P < 0.05 or P < 0.01) by 24.11% and 27.47% on day 20 based on the control group (CG) levels. On day 15, the reductions of FSHR mRNA levels in FRBI-2, FRBI-3 and FRBI-4 were 27.78%, 29.37% and 43.65% (P < 0.05 or P < 0.01), respectively in comparison with CG. ERβ and FSHR protein levels of FRBI-treated mice were gradually decreased as compared to and CG and FSH group. ERβ protein level of FRBI-4 was less than that of CG on day 20 (P < 0.05). On days 15 and 20, estradiol (E 2 ) concentrations of FRBI-2, FRBI-3 and FRBI-4 groups were lower than those of the CG and FSH group (P < 0.05 or P < 0.01)., Conclusions: FRBI could reduce OCT and follicle numbers. A high dose of FRBI (30 mg/kg to 40 mg/kg) could suppress ovarian and follicular development, and attenuate expression levels of ERβ and FSHR mRNAs and proteins in the ovaries, additionally inhibit E 2 production. Therefore, FRBI will possibly be utilized to restrain the carcinogenesis of ovarian cancer by down-regulating overexpression of FSHR and ERβ in the ovaries., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

The population dynamics of annual social hymenoptera such as vespids depend largely on the fertility of the foundresses, which, in turn, is a key factor in the context of biological invasions. The native European hornet Vespa crabro (Vc) and the invasive Asian hornet Vespa velutina (Vv) have generally similar ecological traits, e.g. nesting and feeding habits, although they differ in colony size, which is higher in Vv. Furthermore, in contrast to Vc, Vv is more specialized in its predatory habits, intensively hunting honey bees at the hive. Comparing the morphological and reproductive traits of two closely related species occupying the same ecological niche, one of which is a native species and the other an alien, can help us to gain an understanding of the invasion process. To this end, we here compare reproductive (ovarian size and maturation, fat level, spermatheca size and sperm stock, fecundity) and morphological traits (head size, weight) of the foundresses of these two hornet species. We observed that ovarian maturation began approximately one month earlier in Vv than in Vc, and that the fat level in the former was lower. We found twice the number of sperm in the mated foundresses of Vv than in those of Vc (more than 100 × 10 3 and less than 50 × 10 3 sperm, respectively), in a 16% smaller spermatheca in Vc. Furthermore, the sperm of Vv was found to be 65% shorter than that of Vc. The precocity and higher potential fecundity of Vv queens may have favoured this species over Vc in terms of predatory behaviour, and thereby contributed to its invasiveness., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The objective of the present study was to describe histological development of the European long-snouted seahorse Hippocampus guttulatus, to increase understanding of the biology and physiology of the species. Most vital organs were present in juveniles by the time of their release from the male's pouch. Digestive tract specialization occurred at 89 effective day-degrees (D° eff ), corresponding to 15 days post partum (dpp), with development of the first intestinal loop and mucosal folding. At 118 D° eff (20 dpp), lipids were being mobilized from the liver and oocytes attained the perinuclear stage. The fovea emerged at 177 D° eff (30 dpp), contemporaneous with the shift from pelagic to benthic behaviour in juveniles. At this stage, the most interesting feature was the formation of the second intestinal loop. Male gonads were never observed during the study (from 0 to 354 D° eff ; 0-60 dpp), but the first oogonia were present at 30 D° eff (5 dpp). In 354 D° eff (60 dpp) juveniles, oocytes were observed in a cortical alveoli stage, indicating maturity. Low digestive efficiency was observed at early stages, which was due to a poorly developed gastrointestinal tract and an immature digestive tract prior to 89 D° eff . The present study demonstrates that approximately 89 and 177 D° eff represent two important transitional stages in the early development of H. guttulatus. At a temperature of approximately 19 ± 1°C and an age of 1 month (177 D° eff ), main organs were fully functional, suggesting that the adult phenotype was largely established by that age, with females becoming mature at the age of 2 months (354 D° eff )., (© 2018 The Fisheries Society of the British Isles.)

During early pregnancy, the secretome of both oviduct and uterus serves as exchange medium for signaling factors between embryo and mother and provides the embryo with nutrients. The preimplantation embryo can utilize the fatty acids (FA) therein via direct incorporation into cell membrane lipid bilayers and for energy production via β-oxidation. The FA concentration and composition of the oviduct (OF) and uterine fluid (UF) might be regulated by ovarian hormones to meet the changing needs of the growing embryo. In our study, we analyzed the FA profile of blood plasma (BP) and reproductive fluid samples obtained post mortem from slaughtered mares by gas chromatography mass spectrometry. Cycle stage was determined by visual evaluation of the ovary and measurement of plasma progesterone. No major effect of cycle stage on the FA profile was observed. However, the composition of FA was different between BP and both OF and UF. While linoleic, stearic, oleic and palmitic acid were the four most prevalent FA in both BP and reproductive fluids, the latter contained higher concentrations of arachidonic, eicosapentaenoic and dihomo-γ-linolenic acid. The finding suggests selective endometrial transport mechanisms from plasma into the reproductive fluids or increased endometrial synthesis of selected FA., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Orexins are neuropeptides with pleiotropic functions, involved in the coordination of multiple versatile physiological processes, in particular related to food intake and several aspects of the reproductive process. Their actions are carried out through the bond with the related Orexin 1 (OXR1) and Orexin 2 (OXR2) G-protein-coupled receptors. Studies on the expression of the orexinergic system in the female genital organs are scarce and limited to preovulatory gametogenic follicles and corpora lutea isolated from the rest of the ovary. As the description of only these structures is insufficient to provide a complete picture of the organ, the present study is aimed to give a panoramic view of all the ovarian structures and cells expressing Orexin A (OXA) and its receptors in their original localization. Double labeling immunofluorescent methods, applied on frozen sections of the whole organ in both follicular and luteal phase, were used to highlight the particular distribution and colocalization of the proteins. For a better recognition of cellular morphology and a better distinction between gametogenic (healthy) and atretic follicles, also a single labeling immunolocalization of OXA on formalin fixed paraffin embedded tissues and a TUNEL staining were performed. The results indicate that OXA and its two receptors subtypes are expressed in all the different structures composing the swine ovary, albeit in different ways, in both phases of the ovarian cycle. In general, OXA and OXR2 appear diffusely distributed within "health", proliferating and steroid producing cells, while has granular appearance, being presumably associated to cytoplasmic vesicles, in degenerating cells, independently if apoptotic or not. The immunoreactivity for OXR1, instead, is often associated with the nuclear envelope but it is also detectable, to a lesser extent, diffusely distributed in the cytoplasm of growing or steroid producing cells. When cells undertake the path leading to degeneration, also OXR1 immunoreactivity assumes a granular appearance in the cytoplasm and is colocalized with OXA and OXR2. Different roles for the two receptors in the same cell and a different regulation of their expression remain to be investigated. Their comprehension could help studies of follicle development in pig, as part of in vitro oocyte maturation and fertilization programs in livestock., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Objective: To describe the optimal recumbency for laparoscopic ovariectomy (LapOVE) in dogs., Study Design: Prospective clinical trial., Animals: Sixteen healthy client-owned female dogs undergoing routine single-port LapOVE., Methods: Anesthetized dogs were placed in dorsal recumbency on a tilting operation table. The operating laparoscope was introduced and, after inspection of abdominal organs, directed toward the randomly assigned ovary. Fifteen-second video recordings were taken at each table rotation of 0°, 22.5°, and 45°. LapOVE was performed using the single-port technique. The procedure was repeated on the contralateral side. Blinded observers (3 inexperienced [IO], 3 experienced [EO], 1 expert [EE]) evaluated the recordings postoperatively for visibility of ovary/ovarian bursa, ovario-uterine transition, and other organs., Results: In the 0° position, ovarian structures were rarely visualized (IO, 13/96 [13.5%]; EO, 7/96 [7.3%]; EE, 5/32 [15.6%]). In the 22.5° position, visualization improved significantly for EO (26/96 [27.1%], P < .001). There was a marked improvement in visualization for all groups when the positioning angle was altered to 45° (IO, 62/96 [64.6%]; EO, 67/96 [69.8%]; EE, 25/32 [78.1%]). The differences between 0° and 45° and between 22.5° and 45° were significant for all groups of observers (all P < .001). There was an advantage for ovary visualization when procedures were started with left ovariectomy., Conclusion: The 45° table rotation provided the best ovary visualization, regardless of body side or observer experience. There is evidence that procedures should be started with the left ovary when body side declination is used for organ retraction from the ovaries., (© 2017 The American College of Veterinary Surgeons.)

Research on reproduction and fertility is an essential fundament of management strategies in European wild boar. Concerning the examination of the reproductive tract, methods still lack conformity and comparability. So far the reproductive status of female wild boar has been assessed by ovarian macroscopic examination. In order to find out if this is a valid method, 21 ovaries of 8-34 months old animals covering (early) follicular phase and cyclic inactivity were evaluated microscopically. Number and dimension of follicles as well as the occurrence of atretic follicles were compared and related to age, weight, ovary weight and size of ovary. For this study, 21 ovaries of the right side of each animal were weighed and measured. The diameter of the functional bodies was measured and functional bodies were counted. Size of ovary was calculated by length*width*height (cm 3 ). Macroscopically, follicles were classified into four categories by diameter: F1 (0.2-0.3 cm), F2 (0.4-0.5 cm), F3 (0.6-0.8 cm) and F4 (0.9-1.3 cm). Microscopically, antral follicles of every 10th slice per section of each ovary were counted and classified into three categories by diameter: F0 (≥2 mm), FS (1-2 mm) and FXS (≤1 mm). In follicles with a diameter greater than 2 mm (F0), length and width were measured from the beginning of theca interna. Results showed that age and weight of wild boars were highly correlated (cor = 0.845, p < 0.001). Wild boar of higher age and weight showed higher ovary weight, but this was not significant (p = 0.135 (age), p = 0.329 (weight)). Although the number of macroscopically seen follicles differed widely by animal, microscopically only one ovary showed only small tertiary follicles. Animals with bigger follicles showed a significantly (p = 0.006) smaller number of small and smallest tertiary follicles. The number of tertiary follicles (F0) was not significantly influenced by age or carcass weight (p = 0.460, 0.159). Older and heavier animals showed significantly less FX and FXS (p < 0.001). Comparison of the follicular composition of young wild boar ovaries with domestic pig research allows the conclusion that assessment of cyclic activity and near puberty is possible. However, due to the fact that puberty is a long term biological progress, defining the time of sexual maturity will always be an estimate. Nevertheless, microscopic examination confirmed the macroscopic findings, including follicular dimensions and developmental stage allowing the conclusion that macroscopic evaluation of ovarian functional bodies in wild boar is an easy and reliable way to assess reproductive performance., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In Thalassodrilides cf. briani, the paired ovaries are inconspicuous and polarized structures with developmental gradient of germ cells along their long axis. The about 300 germ cells in the ovary are consolidated into one syncytial cyst and each cell is connected to a common and branched mass of cytoplasm via one stable cytoplasmic bridge. The germ cells differentiate their fate into nurse cells and oocytes. Only one oocyte grows in a given time; it gathers cell organelles and yolk and then it detaches from the gonad. Nurse cells appear to support oocytes development by providing with at least cell organelles. Such observations suggest ovary meroism. T. cf. briani, belonging to limnodriloidin naidids, has the same ovary organization as the representatives of tubificin naidids studied to date. This supports the concept of sister-group relations between Limnodriloidinae and Tubificinae. A similar ovary morphology is also known in several other groups of "microdrile" oligochaetes, which shows that this ovary type is the most widespread among this taxa. Moreover, living in marine or brackish-water sediments, T. cf. briani do not show any significant differences in their ovary structure and oogenesis with freshwater tubificins. This indicates its conservative character independent of the animal life environment., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae) is a widely distributed pest of soft-skinned and stone fruits that is controlled mainly with pesticides. An alternative to the chemical control is the sterile insect technique (SIT), an ecologically friendly method of pest management that could be used against D. suzukii. The objective of the present study was to evaluate the effects of gamma radiation on reproductive sterility, ovarian morphometry, and quality parameters of D. suzukii. Full female sterility was achieved at 75 Gy, while an adequate level of male sterility (99.67%) was obtained at 200 Gy. The ovarian size showed an exponential decay in function of irradiation dose increase. There was no significant influence of irradiation dose on the quality parameters evaluated. Our data suggest that gamma radiation can be recommended to be used in an SIT program for D. suzukii.

To expand our understanding of the roles of thyroid hormones on female reproduction, we induced hypo- and hyper-T rat models to investigate the roles of thyroid hormones on estrous cyclicity, as well as the antioxidative status in the ovaries of rats. In the current study, our data show that hypothyroidism (hypo-T) and hyperthyroidism (hyper-T) led to significantly reduced body weights and ovarain weights and delayed vaginal opening day. For hyper-T, thyroxine (T4), tri-iodothyronine (T3), progesterone (P4) and follicle-stimulating hormone (FSH) were significantly increased, while estradiol (E2) and luteinizing hormone (LH) were significantly decreased. For hypo-T rats, serum levels of total T4 and T3, E2, P4, FSH and LH were significantly increased, while concentrations of E2 and LH were significantly decreased. For ovary morphology, the numbers of secondary and antral follicles were significantly decreased with more atretic antral follicles and less corpora lutea in both hyper- and hypo-T groups. Both hyper-T and hypo-T treatment significantly decreased the expressions of thyroid hormone receptor α1 in the ovary. Hypo-T significantly reduced nitric oxide (NO), total NO synthase (tNOS), inducible NOS and constitutive NOS activities, but hyper-T increased them. For antioxidative parameters, hypo-T and hyper-T treatment significantly increased malondialdehyde (MDA) contents. The activities of both glutathione peroxidase (GSH-Px) and catalase (CAT) significantly decreased in the hypo-T group but increased in the hyper-T group. Total superoxide dismutase (T-SOD) activity was significantly increased in the hyper-T group. In summary, thyroid hormones alter estrous cyclicity and antioxidative status in the ovary of the rat may act through the NOS signaling pathway., (© 2017 Japanese Society of Animal Science.)

We present a robust method called improved-Genome editing via Oviductal Nucleic Acids Delivery (i-GONAD) that delivers CRISPR ribonucleoproteins to E0.7 embryos via in situ electroporation. The method generates mouse models containing single-base changes, kilobase-sized deletions, and knock-ins. The efficiency of i-GONAD is comparable to that of traditional microinjection methods, which rely on ex vivo handling of zygotes and require recipient animals for embryo transfer. In contrast, i-GONAD avoids these technically difficult steps, and it can be performed at any laboratory with simple equipment and technical expertise. Further, i-GONAD-treated females retain reproductive function, suggesting future use of the method for germline gene therapy.

Objective: To explore the therapeutic effects of bone marrow-derived mesenchymal stem cells (BMSCs) on premature ovarian failure (POF) in mice induced by cyclophosphamide (CTX) and the possible mechanisms.
 Methods: Mouse BMSCs were identified through detection of cell surface markers by flow cytometry. The model of mouse POF was induced by intraperitoneal injection of CTX at a dose of 50 mg/kg, once daily for 15 days. BMSCs were transplanted into POF mice at 2×106 cells/mouse by tail veil. The ovarian tissues were collected for HE staining at 7 days after transplantation to observe the changes of ovarian structure and real-time PCR was performed to detect the folliculogenesis gene expression.
 Results: BMSCs showed positive expression of CD29 and CD90 while low expression for endothelial and hematopoietic cell markers CD31 and CD34. The numbers of primodial follicle, primary follicle, secondary follicle and antral follicle were significantly decreased, but the numbers of atretic follicle were significantly increased in CTX induced-POF mice (P<0.05). BMSCs transplantation effectively repaired the structure of damaged ovary. The significant reduction of atretic follicle and significant increase of antral follicle and secondary follicle were observed in ovaries of BMSCs-treated mouse(P<0.05). BMSCs-transplanted mouse ovaries showed the increased mRNA expression levels of Nano3, Nobox, and Lhx8 (P<0.05).
 Conclusion: BMSCs could effectively repair ovarian structure and promote follicle development in CTX-induced POF mouse.

House flies (Musca domestica) are worldwide agricultural pests with estimated control costs at $375 million annually in the U.S. Non-target effects and widespread resistance challenge the efficacy of traditional chemical control. Double stranded RNA (dsRNA) has been suggested as a biopesticide for M. domestica but a phenotypic response due to the induction of the RNAi pathway has not been demonstrated in adults. In this study female house flies were injected with dsRNA targeting actin-5C or ribosomal protein (RP) transcripts RPL26 and RPS6. Ovaries showed highly reduced provisioning and clutch reductions of 94-99% in RP dsRNA treated flies but not in actin-5C or GFP treated flies. Gene expression levels were significantly and specifically reduced in dsRNA injected groups but remained unchanged in the control dsGFP treated group. Furthermore, injections with an Aedes aegypti conspecific dsRNA designed against RPS6 did not impact fecundity, demonstrating species specificity of the RNAi response. Analysis of M. domestica tissues following RPS6 dsRNA injection showed significant reduction of transcript levels in the head, thorax, and abdomen but increased expression in ovarian tissues. This study demonstrates that exogenous dsRNA is specifically effective and has potential efficacy as a highly specific biocontrol intervention in adult house flies. Further work is required to develop effective methods for delivery of dsRNA to adult flies.

Zebrafish, a suitable and widely used teleost fish model in basic biomedical research, displays morphophysiological features of adult gonads that share some commonalities with those of mammalian species. In mammals, gametogenesis is regulated, among several factors, by brain-derived neurotrophic factor (BDNF). This neurotrophin has a well-established role in the developing and adult nervous system, as well as gonads development and functions in vertebrate species. We hypothesize that BDNF has a role also in the gonadal functions of zebrafish. At this purpose, we investigated BDNF and its receptors p75 and TrkB in the ovary and testis of adult zebrafish, kept under laboratory conditions. Our results display (1) the expression of BDNF mRNA and pro-BDNF protein outside of the nervous system, specifically in the ovary and testis; (2) the presence of pro-BDNF in primary oocytes and follicular layer, and p75 in follicular cells; (3) the localization of pro-BDNF in type B spermatogonia, and Sertoli cells in testis. Altogether, these data lead us to consider that BDNF is involved in the gonadal function of adult zebrafish, and mainly in the adult ovary. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 301:140-147, 2018. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

The present study was undertaken to examine the cellular localization and potential steroidogenic role of neuropeptide Y (NPY) in the ovary of the freshwater catfish, Clarias batrachus. NPY-immunoreaction was observed in the follicular cells (granulosa and thecal cells) in the growing ovarian follicles, and the intensity of staining increased steadily from the initiation of follicular development until follicles were fully grown. Thereafter as follicles matured the stain intensity decreased. Positive correlations were found between NPY expression and the ovarian levels of 17β-estradiol, testosterone, and activities of 3β-hydroxysteroid dehydrogenase (3β-HSD) and 17β-hydroxysteroid dehydrogenase (17β-HSD) in the ovary. In vitro NPY treatment stimulated the production of the two steroids and the activities of two enzymes. This is the first report of NPY immunoreactivity at the cellular level in the fish ovary and implicates this orexigenic peptide in the modulation of ovarian steroidogenesis., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Reproductive success is one of the central tenets of conservation management programs, yet the inability to study underlying physiological processes in a minimally-invasive manner and the unpredictable nature of wild animal populations leaves large gaps in our knowledge of factors critical to successful reproduction in wild species. This study integrated ultrasonography of the reproductive tract and analysis of reproductive hormones in 172 northern fur seals (Callorhinus ursinus) to identify intrinsic factors associated with reinitiating embryonic growth at the end of diapause. Within the first 3-4 weeks of active gestation, pregnant fur seals (n = 126) had a larger corpus luteum and fewer antral follicles than non-pregnant fur seals, or those still in diapause (n = 46). This suggests that the conceptus drives changes in ovarian status to convey its presence to the female. Morphological changes in the reproductive tract associated with pregnancy were not reflected in differences in endocrine profiles (estradiol, estrone, progesterone, and relaxin) between pregnant and non-pregnant individuals. Hormone concentrations correlated more strongly with calendar date than with the presence or size of the conceptus, demonstrating that none of these reproductive hormones were reliable markers for early pregnancy diagnosis. Instead, the northern fur seal's long diestrus may serve to reduce the probability of a temporal mismatch between corpus luteum regression and embryo implantation. Indeed, conception rates were high and confirmed rates of pregnancy loss were relatively low (11%). In this study, minimally-invasive ultrasonography was used in wild pinnipeds to detect very early pregnancy (embryonic vesicles >2 mm) in combination with ovarian and endocrine dynamics at the time of embryo implantation, shedding light on mechanisms for maternal recognition of pregnancy. This study is also the first to track whether these same animals carried the embryo to term, by observing fur seals during the birthing season the following year. Data do not support the notion that decreased pregnancy rates or higher pregnancy loss rates are major contributing factors to the northern fur seal's population decline., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The role of thecal glands in the ovary of birds remains controversial. Using transmission electron microscopy and immunohistochemistry, immunohistochemical localisation of cyclooxygenase I and II (COX-1 and COX-2), oestrogen receptor α and β (ER-α and ER-β), androgen receptor (AR) and progesterone receptor (PR), a detailed analysis of the thecal glands was performed. Our ultrastructural studies revealed that the thecal glands of the quail ovary consist of 2 cell types, steroid-producing cells (SPCs) and enclosing cells (ENCs). The SPCs are large, light cells containing a varying number of lipid droplets. Their cytoplasm is characterised by a large amount of smooth endoplasmic reticulum. The ENCs are always located at the periphery of the gland. Some ENCs contain an abundant number of microfilaments, but lipid droplets and dense bodies were rare. Within 1 gland, SPCs with distinct COX-2 immunostaining were interspersed between usually larger numbers of moderately COX-2-positive cells. A completely different staining pattern was observed for COX-1, where the cytoplasm of the ENCs was distinctly immunopositive. The SPCs stained only weakly with antibodies to COX-1. The thecal glands showed distinct reactions for ER-β but only a weak to negative one for ER-α, PR, and AR. Our immunohistochemical and ultrastructural data support our hypothesis that the thecal glands of the quail are involved in steroid hormone and prostaglandin synthesis. The prostaglandins secreted by the thecal glands probably contribute to the ovulation of the follicle first in the hierarchy., (© 2018 S. Karger AG, Basel.)

Background: Food restriction during pregnancy can influence the health of the offspring during the adulthood. The aim of the present study was to examine the effect of maternal food restriction (MFR) on the reproductive performance in female rat offspring from the first (FR1) and second (FR2) generations., Methods: Adult virgin Wistar female rats were given free access to tap water and were fed ad libitum on standard rodent chow, were mated with virgin adult males, and then were randomly divided into two groups: controls (that was fed ad libitum ) and food-restricted group (FR, that was given only 50% of ad libitum food throughout gestation). Their first (FR1) and the second (FR2) generation of offspring were fed ad libitum and sacrificed before puberty and at adulthood. Their ovaries were removed and their histology evaluated by estimating the number of follicles (total and at various stages of folliculogenesis), and the presence of multi-nuclei oocytes and multi-oocyte follicles., Results: Total number of ovarian follicles was lower in FR1 females at week 4 in comparison with controls, while it was not different in FR2 females vs., Controls: The number of the primordial follicle was lower in FR1 and FR2 females vs. controls at both week 4 and at week 8. When compared to the controls, the follicles containing multi-nuclei oocytes were more frequent in ovaries from FR1 and FR2 females at week 4, and higher and lower respectively in ovaries form FR1 and FR2 females at week 8., Conclusion: MFR affects ovarian histology by inducing the development of abnormal follicles in the ovaries in first and second generation offspring. This finding could influence the ovarian function resulting in an early pubertal onset and an early decline in reproductive lifespan.

The ovary is the main organ of the female reproductive system and is essential for the production of female gametes and for controlling the endocrine system, but the complex structural relationships and three-dimensional (3D) vasculature architectures of the ovary are not well described. In order to visualize the 3D connections and architecture of blood vessels in the intact ovary, the first important step is to make the ovary optically clear. In order to avoid tissue shrinkage, we used the hydrogel fixation-based passive CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/ Immunostaining/In situ-hybridization-compatible Tissue Hydrogel) protocol method to clear an intact ovary. Immunostaining, advanced multiphoton confocal microscopy, and 3D image-reconstructions were then used for the visualization of ovarian vessels and follicular capillaries. Using this approach, we showed a significant positive correlation (P <0.01) between the length of the follicular capillaries and volume of the follicular wall.

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in reproductive medicine. In the past, the study of reproductive system development in rodents has been based on one-time histological examination of excised tissues. Recently, with the development of high-resolution transabdominal ultrasound, high-quality sonography can now be performed to evaluate the reproductive organs of rats, allowing a new method for studying the reproductive system. Images were obtained using a high-resolution ultrasonographic system. Gynecological ultrasonography was performed on 28 eight-week-old non-pregnant rats and 5 pregnant Sprague-Dawley rats. We describe how to recognize organs of the reproductive system and associated structures in typical views during different phases of the estrus cycle. Color flow Doppler was used to measure uterine artery blood flow and evaluate uterine blood flow pattern changes during different stages of pregnancy. We have demonstrated that ultrasound exploration is a useful method for evaluating changes in internal reproductive organs. Its use raises the possibility of conducting additional experiments, including medical or surgical procedures, and provides the ability to monitor sonographic changes to internal organs without sacrificing animals.

The structure of the ovary and oogenesis of Poecilia mexicana from an active sulfur spring cave is documented. Poecilia mexicana is the only poeciliid adapted to a subterranean environment with high hydrogen sulfide levels and extreme hypoxic conditions. Twenty females were captured throughout one year at Cueva del Azufre, located in the State of Tabasco in Southern Mexico. Ovaries were processed with histological techniques. P. mexicana has a single, ovoid ovary with ovigerous lamella that project to the ovarian lumen. The ovarian wall presents abundant loose connective tissue, numerous melanomacrophage centers and large blood vessels, possibly associated with hypoxic conditions. The germinal epithelium bordering the ovarian lumen contains somatic and germ cells forming cell nests projecting into the stroma. P. mexicana stores sperm in ovarian folds associated with follicles at different developmental phases. Oogenesis in P. mexicana consisted of the following stages: (i) oogonial proliferation, (ii) chromatin nucleolus, (iii) primary growth, subdivided into: (a) one nucleolus, (b) multiple nucleoli, (c) droplet oils-cortical alveoli steps; (iv) secondary growth, subdivided in: (a) early secondary growth, (b) late secondary growth, and (c) full grown. Follicular atresia was present in all stages of follicular development; it was characterized by oocyte degeneration, where follicle cells hypertrophy and differentiate in phagocytes. The ovary and oogenesis are similar to these seen in other poeciliids, but we found frequent atretic follicles, melanomacrophage centers, reduced fecundity and increased of offspring size., (© 2017 Wiley Periodicals, Inc.)