Ovarian maturation and oogenesis in the blue swimmer crab,Portunus pelagicus (Decapoda: Portunidae)

The study was aimed at understanding the process of reproduction and the changes happening in the ovary of Portunus pelagicus during maturation, which would be useful for its broodstock development for hatchery purposes. For that, tissue samples from different regions of the ovary at various stages of maturation were subjected to light and electron microscopy, and based on the changes revealed and the differences in ovarian morphology, the ovary was divided into five stages such as immature (previtellogenic oocytes), early maturing (early vitellogenic oocytes), late maturing (late vitellogenic oocytes), mature (vitellogenic oocytes), and spent (resorbing oocytes). The ovarian wall comprised of an outermost thin pavement epithelium, a middle layer of connective tissue, and an innermost layer of germinal epithelium. The oocytes matured as they moved from the centrally placed germinal zone toward the ovarian wall. The peripheral arrangement of nucleolar materials and the high incidence of cell organelles during the initial stages indicated vitellogenesis I. Movement of follicle cells toward oocytes in the early maturing stage and low incidence of mitochondria and endoplasmic reticulum in the ooplasm during late vitellogenic stage marked the commencement and end of vitellogenesis II, respectively. Yolk granules at various stages of development were seen in the ooplasm from late vitellogenic stage onwards. The spent ovary had an area with resorbing oocytes and empty follicle cells denoting the end of one reproductive cycle and another area with oogonial cells and previtellogenic oocytes indicating the beginning of the next.     

Ion current activity and molecules modulating maturation and growth stages of ascidian (Ciona intestinalis) oocytes

Electrophysiological techniques were used to study ion currents in the ascidian Ciona intestinalis oocyte plasma membranes during different stages of growth and meiosis. Three stages (A, B, C) of immature oocytes were discriminated in the ovary, with the germinal vesicle (GV) showing specific different features of growth and maturation. Stage A (pre‐vitellogenic) oocytes exhibited the highest L‐type Ca²⁺current activity, and were incompetent for meiosis resumption. Stage B (vitellogenic) oocytes showed Na⁺ currents that remained high during the maturation, up to the post‐vitellogenic stage C oocytes. The latter had acquired meiotic competence, undergoing spontaneous maturation and interacting with the spermatozoon. However, fertilized oocytes did not produce normal larvae, suggesting that cytoplasmic maturation plays a specific role in embryo development. Spontaneous maturation was inhibited at low pH whereas trypsin was able to trigger germinal vesicle breakdown (GVBD) regardless of pH; in addition spontaneous maturation was not affected by removal of follicle cells or by inhibiting junctional communication between oocyte and follicle cells. Taken together these results imply: (i) Ca²⁺ and Na⁺ currents are involved in meiotic progression, growth, and acquisition of meiotic competence; (ii) trypsin‐like molecules may have a role as candidates for providing the physiological stimulus to resume meiosis. Finally, we provide evidence that follicle cells in Ciona are not involved in triggering GVBD as it occurs in other ascidians. Mol. Reprod. Dev. 76: 1084–1093, 2009. © 2009 Wiley‐Liss, Inc.     

Involvement of protein kinase C in the regulation of oocyte maturation in amphibians (Rana dybowskii)

Ovarian oocytes of Rana dybowskii, isolated early in the hibernation period (late autumn), failed to mature, i.e., germinal vesicle breakdown (GVBD), in response to progesterone during in vitro follicle culture. Oocytes collected during the middle hibernation period matured in response to progesterone, whereas those collected late during the hibernation period (close to the breeding season) underwent spontaneous maturation without added hormone (Kwon et al., '89). The maturational response (GVBD) of oocytes, collected at the three stages of hibernation, to protein kinase C (PKC) activation was investigated and compared to that of progesterone stimulation. A phorbol ester, phorbol 12-myristate 13-acetate (TPA) was used for PKC activation. TPA addition to cultured follicles collected during the early or middle period of hibernation induced oocyte GVBD. The incidence of maturation (% GVBD) induced by TPA varied markedly between animals. TPA (10 microM) induced oocyte maturation in the presence or absence of follicle cells. The time course of the TPA-induced maturation was similar to that of progesterone-stimulated maturation (ED50, 7-9 h). TPA also accelerated the onset of maturation of the follicular oocytes exhibiting spontaneous in vitro maturation. Both TPA- and progesterone-stimulated maturation was blocked by treatment with cycloheximide (1 microgram/2 ml), forskolin (9 microM) (an adenylate cyclase stimulator), and verapamil (0.27 mM) (a calcium transport blocker). Treatment of oocytes with a calmodulin antagonist N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) (100 microM) or a PKC inactivator 1-(5-isoquinolinylsulfonyl)-2-methyl-piperazine (H-7) (50 microM) likewise suppressed TPA- or progesterone-induced maturation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dynamics of Oogenesis in a Lower Vertebrate,the Toad Bufo bufo

The patterns of production and growth of oocytes were studied in female toads brought into the laboratory after hibernation and kept at 20°C. The oogenic state of the ovaries was determined from the number and size frequency distribution of the oocytes, and change in oogenic state was followed by comparing ovarian biopsies with the ovaries at autopsy one month later. New oocytes grew within one month to sizes up to 0.16 mm or more in diameter, and were integrated in the existing pool of small oocytes. Production of new oocytes continued for more than one month, perhaps 2–3 months, to increase the pool of oocytes by about 50%. An episode of oogenesis sensu stricto is followed by a resting period, probably lasting for about 1–2 years. Oogenesis was not coordinated with ovulation or recruitment of oocytes to vitellogenic growth and it occurred also in the ovaries of sexually immature toads. Exogenous gonadotropin (hCG) and/or fasting for one month of toads in a good nutritional condition did not affect oogenesis, but no oogenic episodes were observed in the ovaries of starving toads that were depleted of energy reserves. It is concluded that oogenesis and growth of oocytes constitute a dynamic system that is only inadequately described by the widely accepted linear, sequential model of production of eggs in amphibians and teleosts.     

Cell contacts between follicle cells and the oocyte of Helisoma (Mollusca,Pulmonata)

The cell contacts between follicle cells, and follicle cells and oocytes of egg-laying populations of Helisoma duryi and non-egg-laying populations of H. trivcolvis have been studied. Scanning electron microscopy reveals that four to six follicle cells envelop a single developing oocyte. Thin sections and lanthanum impregnations demonstrate apical zonulae adherentes followed by winding pleated-type septate junctions between follicle cells. Gap junctions and septate junctions have been found between follicle cells and vitellogenic oocytes. Freeze-fracture replicas show relatively wide sinuous rows of septate junctional particles, and nemerous large gap junctional particle aggregates on the P-face between vitellogenic oocytes and follicle cells. Septate and gap junctions between immature or nonvitellogenic oocytes and follicle cells are fewer compared to those in vitellogenic oocytes. Similarly, the junctional complexes are less developed in non-egg-laying H. trivolvis compared to those in egg-laying H. duryi. It is possible that intimate interaction between follicle cells and a developing oocyte is necessary for the maturation of the oocyte. The junctional complexes could be involved in the interaction of the follicle cells and the oocyte, and they must disassemble at the onset of ovulation. Rhombic particle arrays and nonjunctional ridges of particles have been found in the basal part of the oolemma.     

Physiological aspects of oogenesis in two species of sticklebacks, Gasterosteus aculeatus L. and Apeltes quadracus (Mitchill)

During May at Woods Hole, female Gasterosteus aculeatus periodically produce clutches of 112±19 eggs with an average diameter of 1·31±0·05 mm. The fish generally have primordial follicles ranging up to 0·56 mm diameter and a clutch of larger follicles undergoing synchronous growth. The size of oocytes within a growing clutch appears to be random within a population, which indicates that recruitment of clutches is not induced by a local environmental event. The largest oocytes within the population of primordial follicles have just begun vitellogenesis but are temporarily arrested. All oocytes within follicles larger than 1·1 mm diameter undergo spontaneous maturation and enlarge to preovulatory size when incubated at 16° C in a simple saline medium. Added deoxycorticosterone can induce similar events in somewhat smaller follicles placed in culture. Thus, in vivo , follicles grow from 0·56 to 1·1 mm diameter by vitellogenesis, and further enlargement is achieved by hydration during steroid-induced maturation. Females carrying follicles in maturational stages also have a new clutch of follicles entering vitellogenic growth from the population of primordial follicles. Injection of human chlorionic gonado-tropin causes a recruitment of follicles into vitellogenesis regardless of the stage of follicles within the growing clutch. One interpretation of these results is that when vitellogenic follicles reach a diameter of 1·1 mm a surge of gonadotropin(s) induces the follicle cells to release steroid, which results in oocyte maturation; the same surge also recruits a new clutch of vitellogenic oocytes. Qualitatively similar results were obtained for Apeltes quadracus ; thus the recruitment phenomenon observed for G. aculeatus may be a general feature among sticklebacks.     

Degeneration of germ line cells in amphibian ovary

Ogielska, M., Rozenblut, B., Augustyńska, R., Kotusz, A. 2010. Degeneration of germ line cells in amphibian ovary. —Acta Zoologica (Stockholm) 91 : 319–327 We studied the morphology of degenerating ovarian follicles in juvenile and adult frogs Rana temporaria, Rana lessonae and Rana ridibunda. Degeneration of primordial germ cells was never observed and was extremely rare in oogonia and early oocytes in a cyst phase in juveniles. Previtellogenic oocytes were rarely affected. Three main types of atresia were identified. In type I (subdivided into stages A–D), vitellogenic oocytes are digested by proliferating follicle cells that hypertrophy and become phagocytic. A – germinal vesicle shrinks, nucleoli fuse, oocyte envelope interrupts, and follicular cells hypertrophy; B – follicular cells multiply and invade the oocyte; C – entire vesicle is filled by phagocytic cells; D – degenerating phagocytes accumulate black pigment. Type II is rare and resembles breakdown of follicles and release of ooplasm. In type III, observed in previtellogenic and early vitellogenic oocytes, ooplasm and germinal vesicle shrink, follicle cells do not invade the vesicle, and condensed ooplasm becomes fragmented. The residual oogonia in adult ovaries (germ patches) multiply, but soon degenerate.     

Dynamics of oogenesis in the tropical anuranRana tigrina (Amphibia: Ranidae) with special reference to vitellogenic cycles in wild-caught and captive flogs

The ovarian cycle ofRana tigrina was analysed by quantifying the developing oocytes (classified into stages on the basis of diameter) and atretic ones at monthly intervals. Stages I to IV represent oocytes in the first growth phase and the remaining ones the vitellogenic or second growth phase. Stages I–III occurred year round but exhibited significant variation in their number. The number of stage II oocytes always dominated the other stages. Recruitment of oocytes to stages IV and V in April marked the initiation of vitellogenic growth in all specimens. Of the 30 to 35% second growth phase oocytes, 25 to 28% reached ovulatory sizes by June. After spawning the ovarian mass declined drastically from 15 to 0.2% of body mass in July. Atresia was maximal (5%) in August. In other months, it was less than 1.5% of the total oocytes. Oogenic episodes occurred in March and July yielding new oocytes. The number of first growth phase oocytes fluctuated from 65 to 95%. The fluctuation was inversely correlated with the second growth phase oocytes indicating a 30 to 35% annual turnover rate of oocytes in the frog. The final egg number/ovarian mass is positively correlated with the snout-vent length as well as body mass of the frogs.R. tigrina produces about 4000 eggs/100g body mass. Further, the mean number of yolky eggs/100 g body mass and the total volume (V) of eggs/frog were highly correlated. Frogs living in captivity produced fewer eggs compared to the wild ones (3594 ± 227 in captivevs 4704 ± 317 in wild frogs). Also, these frogs failed to breed though they showed amplexus with breeding males. Injection of desoxycorticosterone acetate however induced spawning in 4 out of 5 frogs. They released about 3000 eggs each. Captivity seems to mainly impair breeding and to a little extent the vitellogenic growth of oocytes inR. tigrina.     

The annual reproductive pattern of the Antarctic clam, <Emphasis Type="Italic">Laternula elliptica</Emphasis> from Marian Cove,King George Island

The annual reproductive cycle of the Antarctic soft-shelled clam, Laternula elliptica, in Marian Cove, King George Island was studied over a 2-year period from February 1998 to January 2000. Annual changes in the gametogenesis were investigated by measuring the percentage of area occupied by oocytes in a follicle [follicle index (FI)] and the oocyte size. In 1998, the monthly mean FI increased significantly from October to November, peaked in December, and decreased rapidly from December to January. In February and March 1999, degenerated eggs were observed in the spent follicles. Degeneration and resorption of residual eggs by phagocytosis occurred mostly in February and March in both 1998 and 1999, although the resorption process was observed year-round. The histology indicated that complete vitellogenic growth of L. ellpitica at Marian Cove takes at least a year and the clams spawn annually during the austral summer. The ripening and subsequent spawning of clams at Marian Cove in 1998 and 1999 coincided with the algal blooming (September–October 1998 and December and January 1999–2000) suggesting that in coastal Antarctica food supply is a crucial factor that governs gonad maturation and subsequent spawning along with the water temperature.     

Induction of ovarian follicular development in the subadult frogRana tigrina using luteinizing hormone releasing hormone-acetate

In the subadultRana tigrina administration of 2 μg luteinizing hormone releasing hormone-acetate/frog six days a week for 4 weeks in April resulted in the formation of medium (in all 8 frogs) and large sized (in 4 out of 8 frogs) yolky oocytes and, concomitant increases in the oviductal mass. The ovarian and oviductal masses showed a 10-fold increase over the control frogs. In untreated frogs the ovaries were transparent and contained first growth phase oocytes only. The oviducts were also infantile. The pituitary sections were stained using antisera raised in rabbit against the β-subunit of human luteinizing hormone and human follicle stimulating hormone. Immunoreactivity, staining intensity, cytoplasmic granulation and, cell, nuclear and cytoplasmic areas of gonadotrophs (B₂ cells) increased significantly in luteinizing hormone releasing hormone treated frogs. The above findings suggest that pituitary-ovarian axis in the subadultRana tigrina is responsive to luteinizing hormone releasing hormone and that long-term treatment with the hormone induces cytomorphological changes in the gonadotrophs which result in the conversion of inactive cells into secretory cells. This is accompanied by precocious vitellogenic growth of oocytes in the subadult frogs.     

Do fast increasing food conditions promote the midsummer decline of Daphnia galeata?

Ovaries from mature giant red shrimp Aristaeomorpha foliacea were investigated histochemically and ultrastructurally. Four growing stages of the oocytes were distinguished: premeiosis stage, previtellogenetic stage, early vitellogenic stage and late vitellogenic stage. In addition, occasional resorptive oocytes were found. Oogonia and premeiotic oocytes were found in germinative zones. Previtellogenic and vitellogenic oocytes were localized in maturative zones. As vitellogenesis proceeded, oocytes showed a progressive development in the number of lipid droplets as well as in the extension of RER, constituted of dilated cisternae, uniformely scattered throughout the cytoplasm. The RER produced yolk granules and a lampbrush-like substance. The latter was released under the oolemma and constituted a characteristic cortical zone. The oolemma did not develop microvilli or micropinocytotic vesicles to incorporate yolk precursors. Thus, the protein yolk appeared to be of endogenous origin. Few somatic cells were found around the oocytes, but they never gave place to a continuous epithelial layer around oocytes, thus it is not possible to speak of ovarian follicle. The cytoplasm of these mesodermal-oocyte associated cells (MOAC) was characterized by a typical steroidogenic apparatus. Few resorptive immature oocytes were found inside late vitellogenic oocytes. Since the ovaries were packed with late vitellogenic oocytes and the few immature oocytes were hardly detectable, oocyte maturation occurred in a synchronous way.     

Winter Hibernation and UCHL1-p34cdc2 Association in Toad Oocyte Maturation Competence

Currently, it is believed that toad oocyte maturation is dependent on the physiological conditions of winter hibernation. Previous antibody-blocking experiments have demonstrated that toad ubiquitin carboxyl-terminal hydrolase L1 (tUCHL1) is necessary for germinal vesicle breakdown during toad oocyte maturation. In this paper, we first supply evidence that tUCHL1 is highly evolutionarily conserved. Then, we exclude protein availability and ubiquitin carboxyl-terminal hydrolase enzyme activity as factors in the response of oocytes to winter hibernation. In the context of MPF (maturation promoting factor) controlling oocyte maturation and to further understand the role of UCHL1 in oocyte maturation, we performed adsorption and co-immunoprecipitation experiments using toad oocyte protein extracts and determined that tUCHL1 is associated with MPF in toad oocytes. Recombinant tUCHL1 absorbed p34^cdc2, a component of MPF, in obviously larger quantities from mature oocytes than from immature oocytes, and p13^suc1 was isolated from tUCHL1 with a dependence on the ATP regeneration system, suggesting that still other functions may be involved in their association that require phosphorylation. In oocytes from hibernation-interrupted toads, the p34^cdc2 protein level was significantly lower than in oocytes from toads in artificial hibernation, providing an explanation for the different quantities isolated by recombinant tUCHL1 pull-down and, more importantly, identifying a mechanism involved in the toad oocyte’s dependence on a low environmental temperature during winter hibernation. Therefore, in toads, tUCHL1 binds p34^cdc2 and plays a role in oocyte maturation. However, neither tUCHL1 nor cyclin B1 respond to low temperatures to facilitate oocyte maturation competence during winter hibernation.     

Histological investigation on the ovarian cycle of the bluefin tuna in the western and central Mediterranean

The histological analysis of eastern Atlantic bluefin tuna Thunnus thynnus ovaries caught from February to September 1999–2000, made it possible to distinguish the presence of seven oocyte developmental stages and allowed the characterization of six time-dependent ovary maturity stages. The ovaries of mature (fork length, L _F ≥ 110 cm) bluefin tuna were non-active from August (spent period) to March (quiescent period) when they contained only perinucleolar-stage oocytes. Ovary development started in April to early May (recrudescent period) with the appearance of oocytes at the lipid stage. Vitellogenesis appeared in mid-May (ripening period) and post-vitellogenesis occurred in late May to mid-June (pre-spawning period). In late June to early July, hydrated oocytes, a sign of imminent spawning, were found only in specimens caught in Balearic waters. Females ranging between 100 and 110 cm L _F, captured during the recrudescent and ripening periods, had the largest oocytes at the lipid stage, most of which were degenerating. An extensive vitellogenic atresia was observed in the ovaries of five females caught during the spawning period in non-spawning areas.     

Role of temperature in regulation of ovarian cycle in bull frog Rana tigerina

Effect of temperature on the ovarian cycle was studied in R. tigerina by exposing them to (1) constant low (22 degrees C) temperature during preparatory (active vitellogenic growth) phase (March-May) when the mean ambient temperature ranged from 26 degrees-28 degrees C and (2) to constant high (30 degrees +/- 1 degrees C) temperature during postbreeding regression phase (August-November) when the mean ambient temperature ranged from 22 degrees-26 degrees C. The ovaries of initial controls (biopsy samples taken prior to the commencement of the experiment) in March contained only first growth phase (FGP) oocytes with a maximum size range of 361-480 microns in diameter. In the frogs exposed to constant low temperature for 2 months, only 7% of FGP oocytes were recruited to second growth phase (SGP) with a mean largest diameter of 631 microns compared to 31% large SGP oocytes with a mean diameter of 1114 microns in the frogs collected from natural fields. The number of atretic follicles (AF) was lower and fat body weights were significantly higher in low temperature exposed frogs. The exposure of the frogs to constant high temperature during postbreeding months caused an increase in the mean diameter and number of large FGP oocytes, numerical increase in AF and decrease in fat body weights over corresponding controls maintained at room temperature. The pituitary gonadotrophs of these frogs showed stimulatory changes such as increase in cell size and appearance of secretory granules in the cytoplasm. The results suggest that in R. tigerina high temperature stimulates oocyte growth while low temperature retards it.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of gonadotropins on oocyte maturation and progesterone production by porcine ovarian follicles cultured in vitro

Effects of gonadotropins on the maturation of isolated oocytes and production of progesterone by porcine ovarian follicles from gonadotropin treated gilts have been studied in vitro. The addition of gonadotropins (2 I. U./ml, PMSG, HGC or 2 mg/ml FSH) to the culture medium resulted in increasing the number (84 - 90 %) of isolated oocytes which reached metaphase II. Expansion of the whole cumulus mass was observed only in media containing PMSG, whereas FSH or HCG alone did not cause these marked changes in the cumulus cells. Denudation of the eggs prior to culture gave no significant differences in the maturation rates between oocytes cultured in media with or without gonadotropins. In vitro maturation of follicle-enclosed oocytes took place only in HCG treated animals. Removing the ovary at 15 or 60 minutes after intravenous HCG administration induced oocyte maturation only in 22% and 17% respectively. A sharp increase in the number of oocytes which resume meiosis during follicle culture was observed 4 hours after HCG injection (84 %) and all of the oocytes of the gilts ovariectomized at 8 hours after HCG injection matured during the culture period. The progesterone production of isolated follicles from control gilts (only PMSG injected) increased slowly during a 96-hour culture period (from 48 to 240 ng progesterone/follicle), whereas the secretion of progesterone was drastically increased after a 15 minute interval between HCG injection and ovariectomy (from 42 to 950 ng progesterone/follicle). Follicles removed 24 hours after HCG injection showed a further increase in steroid production (2000 ng progesterone/follicle) and consistently secreted large amounts of progesterone during the culture period.     

Meiotic competence in horse oocytes: interactions among chromatin configuration, follicle size, cumulus morphology, and season

Horse oocytes were collected from an abattoir over a 15-mo period. After classification of follicle size and cumulus morphology, oocytes were either fixed immediately (0 h) or matured in vitro (24 h). There was no effect of season on the number of antral follicles present on the ovaries, or on oocyte maturation rate for any class of oocyte. The proportion of oocytes having condensed chromatin at 0 h increased with increasing follicle size. The oocyte maturation rate also increased with follicle size, and for follicles 相似文献   

The ability to achieve meiotic maturation in the dog oocyte is linked to glycolysis and glutamine oxidation

We tested the hypothesis that meiotic competence of dog oocytes is tightly linked with donor follicle size and energy metabolism. Oocytes were recovered from small (<1 mm diameter, n = 327), medium (1–<2 mm, n = 292) or large (≥2 mm, n = 102) follicles, cultured for 0, 24, or 48 hr, and then assessed for glycolysis, glucose oxidation, pyruvate uptake, glutamine oxidation, and nuclear status. More oocytes (P < 0.05) from large follicles (37%) reached the metaphase‐II (MII) stage than from the small group (11%), with the medium‐sized class being intermediate (18%; P > 0.05). Glycolytic rate increased (P < 0.05) as oocytes progressed from the germinal vesicle (GV) to MII stage. After 48 hr of culture, oocytes completing nuclear maturation had higher (P < 0.05) glycolytic rates than those arrested at earlier stages. GV oocytes recovered from large follicle oocytes had higher (P < 0.05) metabolism than those from smaller counterparts at culture onset. MII oocytes from large follicles oxidized more (P < 0.05) glutamine than the same stage gametes recovered from smaller counterparts. In summary, larger‐sized dog follicles contain a more metabolically active oocyte with a greater chance of achieving nuclear maturation in vitro. These findings demonstrate a significant role for energy metabolism in promoting dog oocyte maturation, information that will be useful for improving culture systems for rescuing intraovarian genetic material. Mol. Reprod. Dev. 79: 186–196, 2012. Published 2011. This article is a U.S. Government work and is in the public domain in the USA.     

Risks of hormonally active pharmaceuticals to amphibians: a growing concern regarding progestagens

Most amphibians breed in water, including the terrestrial species, and may therefore be exposed to water-borne pharmaceuticals during critical phases of the reproductive cycle, i.e. sex differentiation and gamete maturation. The objectives of this paper were to (i) review available literature regarding adverse effects of hormonally active pharmaceuticals on amphibians, with special reference to environmentally relevant exposure levels and (ii) expand the knowledge on toxicity of progestagens in amphibians by determining effects of norethindrone (NET) and progesterone (P) exposure to 0, 1, 10 or 100 ng l⁻¹ (nominal) on oogenesis in the test species Xenopus tropicalis. Very little information was found on toxicity of environmentally relevant concentrations of pharmaceuticals on amphibians. Research has shown that environmental concentrations (1.8 ng l⁻¹) of the pharmaceutical oestrogen ethinylestradiol (EE₂) cause developmental reproductive toxicity involving impaired spermatogenesis in frogs. Recently, it was found that the progestagen levonorgestrel (LNG) inhibited oogenesis in frogs by interrupting the formation of vitellogenic oocytes at an environmentally relevant concentration (1.3 ng l⁻¹). Results from the present study revealed that 1 ng NET l⁻¹ and 10 ng P l⁻¹ caused reduced proportions of vitellogenic oocytes and increased proportions of previtellogenic oocytes compared with the controls, thereby indicating inhibited vitellogenesis. Hence, the available literature shows that the oestrogen EE₂ and the progestagens LNG, NET and P impair reproductive functions in amphibians at environmentally relevant exposure concentrations. The progestagens are of particular concern given their prevalence, the range of compounds and that several of them (LNG, NET and P) share the same target (oogenesis) at environmental exposure concentrations, indicating a risk for adverse effects on fertility in exposed wild amphibians.     

A study of ovarian follicular kinetics, oviduct, fat body, and liver mass cycles in laboratory-maintained Rana cyanophlyctis in comparison with wild-caught frogs.

Ovarian follicular dynamics and fluctuations in fat body, oviducal, and liver masses were studied in captive Rana cyanophlyctis in comparison with wild-caught frogs, sampled at monthly intervals over a period of 12 months. In both the captive and wild-caught frogs first growth phase (FGP) and second growth phase (SGP) or vitellogenic oocytes were produced throughout the period examined; however, changes in ovarian and oviducal weights were less marked in the former group. In the captive frogs SGP oocyte production was reduced by 50%, and, maximum ovarian weight and SGP oocyte number were attained 2-3 months earlier than in wild-caught controls. The FGP oocyte pool in laboratory-maintained frogs, however, was comparable with that of the corresponding wild-caught frogs. Captivity caused a threefold increase in atresia and reduced the number of oocytes reaching SGP. The depletion of fat stores in fat bodies during the later phases of captivity suggests that the deposition of lipids into oocytes (for SGP) was given priority over storage in the fat bodies. The low oviducal weights of captive frogs was correlated with a reduced number of SGP oocytes, which are the source of estrogen. On the other hand, liver weight remained high, indicating adequate hepatic vitellogenin synthesis. Possible reduction in its output was not detected, possibly due to the reduced number of follicles reaching SGP. The findings indicate that stress of captivity decreases gonadotrophins and estrogen levels. Oviducal growth is reduced in captive frogs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ultrastructural study of oogenesis in Phoronopsis harmeri (Phoronida)

Temereva, E.N., Malakhov, V.V. and Yushin, V.V. 2011. Ultrastructural study of oogenesis in Phoronopsis harmeri (Phoronida). —Acta Zoologica (Stockholm) 92 : 241–250. The successive stages of oogenesis in Phoronopsis harmeri were examined by electron microscopy methods. During the oogenesis, each oocyte is encircled by vasoperitoneal (coelomic) cells forming a follicle. The previtellogenic oocytes are small cells which accumulate ribosomes for future synthesis; their cytoplasm contains characteristic clusters of mitochondria and osmiophilic particles resembling a germ plasm of other metazoans. The cytoplasm of the vitellogenic oocytes includes numerous mitochondria, cisternae of the rough endoplasmic reticulum, Golgi bodies and annulate lamellae. The synthesis of three types of inclusions was observed: strongly osmiophilic granules (lipid droplets) as a prevalent component, distinctly larger granules surrounded by membrane (proteinaceous yolk) and numerous large vesicles with pale flocculent content. No inclusions which could be unequivocally interpreted as the cortical granules were detected. The surface of the vitellogenic oocytes is covered by microvilli which increase in number and length during development. The oogenesis in Phoronida may be interpreted as follicular because of close association of oocytes with the vasoperitoneal tissue. However, well‐developed synthetic apparatus together with a strongly developed microvillous surface and absence of endocytosis indicate a clear case of autosynthetic vitellogenesis. Thus, in phoronids, there is a combination of simply developed follicle and autosynthesis that, apparently, is plesiomorphic character.