Occurrence of somatic cells within the spermatic cysts of demosponges: a discussion of their role

During ultrastructural studies on the spermatogenesis of two demosponges, Raspaciona aculeata and Petrosia ficiformis, somatic cells were detected within their spermatic cysts. In R. aculeata large, amoeboid somatic cells, presumably derived from follicle cells, were found inside cysts filled with secondary spermatocytes. These cells were actively phagocytosing spermatocytes. Such phagocytosis could be directed to eliminate aberrant spermatogenic cells, maintain appropriate cell number within the cysts, or recapture energetic reserves originally allocated to the reproductive process via phagocytosis of spermatocytes. In P. ficiformis, somatic round cells were observed only after the spawning event, phagocytosing unspawned sperm in nearly emptied spermatic cysts. Unlike in R. aculeata, these cells were more similar to archaeocytes (common cells of the demosponge mesohyl with phagocytic activity) than to follicle cells. Phagocytosis of spermatogenic cells and unspawned spermatozoa by somatic cells from the wall of the testes is a well-known process in vertebrates and many invertebrates. Occurrence of somatic cells in the spermatic cysts of sponges, whose function appears to be partially analogous to that of Sertoli cells, reveals that sponges possess cellular mechanisms to regulate testis functioning that are equivalent to those in higher metazoans.     

Gonadal development and non-functional protogyny in a coral-reef damselfish, Dascyllus albisella Gill

All gonads of the Hawaiian dascyllus Dascyllus albisella , irrespective of the final sex of individuals, developed an ovarian lumen and primary-growth-stage oocytes after an initially undifferentiated state. From this ovarian state or from more differentiated ovaries, some gonads redifferentiated into testes. None of 117 individuals examined had a gonad containing degenerating vitellogenic oocytes and proliferating spermatogenic tissue. Eleven individuals had gonads containing degenerating cortical-alveolus-stage oocytes and developing spermatogenic tissue. The size of these individuals overlapped with the female size range in which the majority of the females were still in the middle of the maturation process. They were absent from the larger size range where the majority of females had vitellogenic oocytes. This indicated that the transition toward maleness is likely to have occurred after the onset of cortical-alveolus stage, but before final oocyte maturation and spawning as females. Therefore the protogynous pattern of gonadal development was non-functional. There was no dimorphism in the sperm duct configuration, and all the testes were secondary testes reported for diandric, protogynous species with undelimited gonads. Very early development of an ovarian lumen appeared to have resulted in a secondary-male configuration in all testes, although redifferentiation into males appeared to have occurred before sexual maturity and spawning as females.     

Germinal epithelium, folliculogenesis, and postovulatory follicles in ovaries of rainbow trout, Oncorhynchus mykiss (Walbaum, 1792) (Teleostei, protacanthopterygii, salmoniformes)

The rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), is a salmoniform fish that spawns once per year. Ripe females that had ovulated naturally, and those induced to ovulate using salmon gonadotropin-releasing hormone, were studied to determine whether follicles were forming at the time of spawning and to describe the process of folliculogenesis. After ovulation, the ovaries of postspawned rainbow trout were examined histologically, using the periodic acid-Schiff procedure, to stain basement membranes that subtend the germinal epithelium and to interpret and define the activity of the germinal epithelium. After spawning, the ovary contained a few ripe oocytes that did not ovulate, numerous primary growth oocytes including oocytes with cortical alveoli, and postovulatory follicles. The germinal epithelium was active in postspawned rainbow trout, as determined by the presence of numerous cell nests, composed of oogonia, mitotic oogonia, early diplotene oocytes, and prefollicle cells. Cell nests were separated from the stroma by a basement membrane continuous with that subtending the germinal epithelium. Furthermore, follicles containing primary growth oocytes were connected to the germinal epithelium; the basement membrane surrounding the follicle joined that of the germinal epithelium. After ovulation, the basement membrane of the postovulatory follicle was continuous with that of the germinal epithelium. We observed consistent separation of the follicle, composed of an oocyte and surrounding follicle cells, from the ovarian stroma by a basement membrane. The follicle is derived from the germinal epithelium. As with the germinal epithelium, follicle cells derived from it never contact those of the connective tissue stroma. As with epithelia, they are always separated from connective tissue by a basement membrane.     

Preliminary results of the in vivo studies on ovarian resorption in carp (Cyprinus carpio L.)

Oocyte resorption in the ovaries of sexually mature carp, which did not spawn, was investigated using a method of sampling ovaries in vivo . A 2-year study revealed that a portion of the oocytes had undergone a slow resorption while the basic mass of the ovaries, throughout the experiment, consisted of the oocytes similar to those of stage IV of sexual maturity. At the same time no growth of the oocytes of the earlier maturation stages was found. The condition of the ovaries in the females studied indicated their complete infertility, at least for three successive spawning periods.     

RNA synthesis in the ovary and oocytes of the starfish

Qualitative studies on the in vitro uptake and incorporation of tritiated uridine into RNA of the somatic and germinal elements of the starfish ovary were carried out prior to and during hormone-induced oocyte maturation and spawning.Autoradiography of nonhormone-treated ovaries indicated that the outer ovarian wall contained the highest concentration of label, with lesser amounts in the follicle cells and least in the oocytes. Oocytes and follicle cells localized at the periphery of the ovary were labeled first, and both cells became progressively labeled throughout the ovary with time; the label first appeared localized in the nucleolus of the oocyte.Sucrose gradient analysis of the separated cellular components of prelabeled hormone-treated ovaries indicated that RNA synthesis occurred in all segments of the ovary and that the spawned oocyte fraction was the least active. Synthesis of ribosomal RNA was detectable after a lag period of approximately 4 hr. Oocytes incubated in ³H-uridine during and subsequent to 1-methyladenine-induced spawning and maturation synthesized 15–19 S and low molecular weight RNA but not ribosomal RNA. Synthesis of the 15–19 S RNA was inhibited with ethidium bromide and to a limited extent by actinomycin D. Isolated mitochondrial fractions contained most of the labeled 15–19 S RNA. These data suggest the mitochondrial origin of most, if not all, of this intermediate-weight RNA. On the basis of these studies, it appears that starfish oocytes and follicle cells are metabolically active at the transitional period from growth to maturational stages in oocytes. Synthesis of RNA furthermore apparently continues in the cytoplasm subsequent to germinal vesicle breakdown and spawning.     

Sperm Storage Structures in the Ovary of Helicolenus dactylopterus dactylopterus (Teleostei: Scorpaenidae): an Ultrastructural Study

The ultrastructure of ovarian sperm storage of Helicolenus dactylopterus dactylopterus is described, before and after the spawning period. The spermatozoa remain inside cryptal structures that are situated in the interlamellar gaps and are connected to the ovarian lumen by a duct. This complex forms a highly specialised structure. During the long storage period, crypts are richly vascularised. Their surrounding simple epithelia have intercellular junctions that may serve to protect the spermatozoa from the female immune system. At the moment during which insemination of mature oocytes occurs, the sperm may be expelled from cryptal structures by means of a spasmodic contraction. During the post spawning period, residual spermatozoa that remain in the crypts are eliminated by cryptal phagocytes. At the end of the process the crypts contain only an amorphous material.     

Immunolocalization of the sea urchin sperm receptor in eggs and maturing ovaries

The sperm receptor from Strongylocentrotus purpuratus eggs is a high molecular weight proteoglycan-like molecule that inhibits fertilization species-specifically in a competition bioassay. A preparation of highly active sperm receptor that contained one major N-terminal sequence was used to generate polyclonal antibody in rabbits. This antibody species-specifically inhibited fertilization at low concentrations without interfering with fertilization envelope elevation. On immunofluorescence microscopy, the antibody recognized determinants on the mature egg cell surface and in the cortical granules just beneath the surface. Preabsorption of the antibody with the calcium-soluble fraction of the exudate from cortical granules rendered the antibody specific for the cell surface in mature eggs and still able to inhibit fertilization at the same concentrations as before treatment with cortical granule exudate. With antibody preabsorbed with cortical granule and by counting antibody-gold particles viewed by electron microscopy, sperm receptor was almost undetectable on the cell surface of immature oocytes in preseason ovaries, present on the cell surface and intracellularly in immature oocytes of ovaries collected at the beginning or at the height of the spawning season, and present only on the cell surface of mature oocytes in the lumen of the ovaries. Our results indicate that receptor is synthesized early in oogenesis and is rapidly moved to the egg cell surface.     

Extrafollicular mediation of oocyte maturation by radial nerve factor in starfish Pisaster ochraceus

In starfish ovaries follicle cells that envelop each oocyte are thought to mediate the production of a maturation inducing substance (MIS), identified as 1-methyladenine, that induces maturation and spawning of oocytes after exposure to a gonadotropic substance secreted by the radial nerve (RNF). Studies were carried out to assess the possible role of extrafollicular cells within the ovarian wall in mediating this signal transduction process in the ovary of Pisaster ochraceus. Oocyte maturation and spawning occurred following the addition of RNF to intact ovarian tissue in vitro whereas no maturation occurred following the addition of RNF to germinal vesicle (GV) oocytes or GV oocytes surrounded by follicle cells. In contrast, oocyte maturation occurred when small ovarian wall fragments, lacking mature follicles, were incubated with GV oocytes and RNF. Neither actinomycin D nor cycloheximide altered RNF induction of oocyte maturation in the presence of the ovarian wall tissue whereas preheating (boiling water for 5 min) the tissue obliterated its response to RNF. Non-ovarian tissues failed to produce MIS in response to RNF. Results suggest that ovarian components other than the follicle cells that envelop fully grown immature oocyte are responsive to RNF and represent a significant and previously unrecognised intra-ovarian source of MIS.     

Morphology and ultrastructure of the somatic cells in Astacus leptodactylus ovary

We defined the somatic environment in which female germinal cells develop, and performed ultrastructural analyses of various somatic cell types, with particular reference to muscle cells and follicle cells, that reside within the ovary at different stages of oogenesis. Our findings show that ovarian wall of the crayfish is composed of long muscle cells, blood cells, blood vessels and hemal sinuses. The follicle and germinal cells lie within a common compartment of ovarian follicles that is defined by a continuous basal matrix. The follicle cells form branching cords and migrate to surround the developing oocytes. A thick basal matrix separates the ovarian interstitium from ovarian follicles compartment. Transmission electron microscopy shows that inner layer of basal matrix invaginates deeply into the ovarian compartment. Our results suggest that before being surrounded by follicle cells to form follicles, oogonia and early previtellogenic oocytes reside within a niche surrounded by a basal matrix that separates them from ovarian interstitium. We found coated pits and coated vesicles in the cortical cytoplasm of previtellogenic and vitellogenic oocytes, suggesting the receptor mediated endocytosis for transfer of material from the outside of the oocytes, via follicle cells. The interstitial compartment between the inner muscular layer of the ovarian wall and the basal matrix of the ovarian follicle compartment contains muscle cells, hemal sinuses, blood vessels and blood cells. Granular hemocytes, within and outside the vessels, were the most abundant cell population in the ovarian interstitium of crayfish after spawning and in the immature ovary. J. Morphol. 277:118–127, 2016. © 2015 Wiley Periodicals, Inc.     

In vitro inhibition of oocyte and follicular maturation and spawning in starfish (Asterias forbesi) by 2-4-dinitrophenol

The in vitro effects of 2-4-dinitrophenol (DNP) on spawning and follicular and oocyte maturation in starfish ovaries and its various cellular components were investigated. Spawning and oocyte and follicular maturation induced by starfish gonadotropin radial nerve factor (RNF) in isolated ovarian fragments were all inhibited by appropriate doses of DNP. DNP inhibits processes which occur shortly after addition of the gonadotropin; in ovarian fragments insensitivity to DNP inhibition occurred shortly after addition of RNF but prior to initiation of spawning. Spontaneous follicular and oocyte maturation which occurred following release of ovarian follicles into sea water was prevented by DNP. In non-spontaneously maturing follicles released from the ovary, DNP inhibited both follicle and oocyte maturation induced by the secondary stimulator of spawning and maturation, 1-methyladenine (1-MA). DNP also inhibited 1-MA induced meiotic maturation in isolated immature oocytes incubated in the absence of follicle cells. Inhibition of oocyte maturation was not associated with inhibition of 3H-1-MA incorporation by isolated oocytes. Immature oocytes incubated in the presence of DNP underwent maturation following washing and subsequent exposure to 1-MA. Immature oocytes initially exposed to both 1-MA and DNP, however, showed decreased maturation responsiveness following washing and re-exposure to 1-MA. The results suggest that the inhibitory effects of DNP on spawning and oocyte maturation are the result of direct effects on the oocytes and possibly other cells and tissues within the ovary.     

In vitro Inhibition of Oocyte and Follicular Maturation and Spawning in Starfish (Asterias forbesi) by 2-4-Dinitrophenol

The in vitro effects of 2-4-dinitrophenol (DNP) on spawning and follicular and oocyte maturation in starfish ovaries and its various cellular components were investigated. Spawning and oocyte and follicular maturation induced by starfish gonadotropin radial nerve factor (RNF) in isolated ovarian fragments were all inhibited by appropriate doses of DNP. DNP inhibits processes which occur shortly after addition of the gonadotropin; in ovarian fragments insensitivity to DNP inhibition occurred shortly after addition of RNF but prior to initiation of spawning. Spontaneous follicular and oocyte maturation which occurred following release of ovarian follicles into sea water was prevented by DNP. In non-spontaneously maturing follicles released from the ovary, DNP inhibited both follicle and oocyte maturation induced by the secondary stimulator of spawning and maturation, 1-methyladenine (1-MA). DNP also inhibited 1-MA induced meiotic maturation in isolated immature oocytes incubated in the absence of follicle cells. Inhibition of oocyte maturation was not associated with inhibition of ³H-1-MA incorporation by isolated oocytes. Immature oocytes incubated in the presence of DNP underwent maturation following washing and subsequent exposure to 1-MA. Immature oocytes initially exposed to both 1-MA and DNP, however, showed decreased maturation responsiveness following washing and re-exposure to 1-MA. The results suggest that the inhibitory effects of DNP on spawning and oocyte maturation are the result of direct effects on the oocytes and possibly other cells and tissues within the ovary.     

Seasonal cycles of gonadal development and plasma sex steroid levels in Epinephelus morio, a protogynous grouper in the eastern Gulf of Mexico

In a field population of the protogynous red grouper Epinephelus morio in the eastern Gulf of Mexico, females with oocytes at all stages of development were collected during the spawning season suggesting that several batches of oocytes may be released over the spawning period. Plasma oestradiol (E₂) levels were highest in ripe females whose gonads contained both cortical alveoli and vitellogenic oocytes during the breeding season. Males were still spermiating as late as August, although levels of androgens 11-ketotestosterone (11-KT) and testosterone (T) had declined from their peaks in March. A few red grouper with either perinucleolar or cortical alveoli stage oocytes were undergoing sex change both during and after the spawning period. Low levels of E₂, T and 11-KT were detected in transitionals. Proliferation of male tissue was not restricted to any specific area of the gonad but occurred in pockets within the ovarian lumen. The sequence of an increase in gonial cells along the periphery of the lamellae, increase in interstitial tissue, degradation of female elements, and formation of a sperm duct seemed to be concurrent with spermatocyte proliferation and the process of preparing the gonad to function as a testis.     

Renewal of the gonads in Styela clava (Urochordata: Ascidiacea) as revealed by autoradiography with tritiated thymidine

DNA-synthesizing cells in the gonads of the ascidian Styela clava were labeled with tritiated thymidine and detected with autoradiography. In the testis, spermatogonia and primary spermatocytes are labeled after 1 hr. Labeled spermatozoa occur in the lumen of the testis follicles after 10 days and in the sperm ducts after 20 days. In the ovary, only germ cells (oogonia and pre-leptotene primary oocytes) and follicle cells are labeled after 1 hr. By 60 days, oocytes with basophilic cytoplasm (15–65 μ in diameter) are labeled; test cells embedded in larger eosinophilic oocytes (150 μ in diameter) are also labeled. Germ cells give rise to both oocytes and follicle cells. Through continued cell division, follicle cells give rise to test cells.     

A histological description of ovarian recrudescence in two Labeo victorianus populations

The ovaries of Labeo victorianus are paired organs situated in the peritoneal cavity and suspended on either side of the midline by a mesovarium. A capsule, composed of dense, regularly-arranged collagen and elastic fibres mixed with a few smooth muscle cells, enclosed the ovaries and gave off connective tissue septa, forming the ovigerous lamellae, which contained germ and follicle cells. Eight discrete stages of recrudescence were identified: oogonia, chromatin nucleolar oocytes, perinucleolar oocytes, primary yolk vesicle oocytes, secondary yolk vesicle oocytes, tertiary yolk vesicle oocytes, post-ovulatory follicles and atretic oocytes. Ovulation seemed to be synchronised with the onset of rainfall, with some deviations in the Sio River population. Gonadosomatic index variation followed a bimodal pattern, with maxima between January–February and between September–October for both populations. The same pattern was exhibited for both rainfall and water levels at the two study sites. Successful ovulation was followed by the formation of post-ovulatory follicles and Type I atresia, while failed spawning was characterised by Type II atresia. Clearance of post-ovulatory follicles was by phagocytosis and formation of melanomacrophage centres. There were variations in post-ovulatory changes between the two populations. Reproductive patterns in the Kagera River population conformed to the 'norm' in African labeines of the synchronisation of spawning with rainfall. Slight deviations from this pattern were, however, observed in the Sio River population where spawning occurred prior to the onset of rainfall.     

Influence of follicle size on the penetrability of immature pig oocytes for homologous in vitro penetration assay

In order to improve the performance of homologous in vitro penetration (hIVP) assays using immature oocytes to assess the penetrating ability of boar sperm, the present study was designed to evaluate the influence of oocyte and follicle size on the penetrability of immature pig oocytes obtained from slaughterhouse ovaries. Nonatretic antral follicles were isolated, measured with a computerized image analysis system and grouped according to their diameter: Group 1 (0.40-0.99 mm), Group 2 (1.00-2.19 mm), Group 3 (2.20-2.79 mm), and Group 4 (2.80-6.50 mm). After sperm coincubation and before penetrability evaluation, the immature oocytes were classified into four size categories according to their diameter excluding zona pellucida: <105, 105-109, 110-114, and > or =115 microm. As regards follicle size, the highest viability and penetrability were obtained with oocytes from follicles >2.20 mm (P>0.05). Regarding oocyte size, significant differences (P<0.05) were observed for all parameters evaluated between oocytes with a diameter above or below 110 microm. However, our results revealed that such differences were due to follicle size rather than oocyte diameter, since oocytes with the same diameter but from different follicle size groups showed different penetration rates. With increasing follicle size, the percentage of penetrated oocytes increased (P<0.05). Finally, our results showed that the greater penetrability of immature oocytes from larger follicles is not due to variations in the thickness of the zona pellucida. There were no significant differences in zona pellucida thickness between oocytes from the four follicular size groups. In summary, these results indicate that follicle size directly affects the penetrability of immature pig oocytes used in hIVP.     

Penetration by bull spermatozoa into the zona pellucida of dead bovine oocytes recovered from frozen-thawed ovaries

The present study was carried out to determine if the zona pellucida of dead bovine oocytes obtained from ovaries stored at -196 degrees C could be used to assess penetrability of capacitated bull spermatozoa. Follicular oocytes were recovered from bovine ovaries which were frozen slowly in a box containing dry ice, plunged into liquid nitrogen, and thawed at 37 degrees C. The dead oocytes were inseminated with various concentrations of spermatozoa preincubated for 0 to 4 h. Sperm penetration rates of the dead oocytes were significantly altered by sperm concentration and preincubation time. Dead and living oocytes matured in vitro (control) gave similar patterns of penetrability based on sperm preincubation time. When sperm concentration was increased, the rate of multiple sperm penetration into the dead oocytes also increased significantly, but the rate of penetration into living oocytes did not alter significantly. All dead oocytes from ovaries stored at -196 degrees C for 1 d to 3 mo were penetrated at similar rates by spermatozoa preincubated for 1-h. Thus, we conclude that dead follicular oocytes recovered from frozen ovaries are useful for the assessment of sperm capacitation and/or the acrosome reaction in cattle.     

The effect of triiodothyronine on oocyte maturation in the starred sturgeon following exposure to low temperatures and female reservation

A sharp cooling of the sevryuga females as well as their reservation at the spawning temperatures suppress the ability of oocytes coated by follicle membranes to mature under the effect of hypophyseal gonadotrophic hormones in vitro and in vivo. At the same time the oocytes from these females in both the types of experiments are able to mature in vitro in the Ringer solution with progesterone. The injection of triiodthyronine to cooled or reserved females restores the ability of their oocytes to mature under the effect of hypophyseal gonadotrophic hormones both in vivo and in vitro. Such oocytes in our experiments undergo normal development upon fertilization. The thyroid gland hormone (triiodthyronine) influences thus, the ability of follicle epithelium to respond to the effect of hypophyseal gonadotrophic hormones by stimulation of oocyte maturation; this influence of triiodthyronine appears to be indirect. Some practical aspects of this problem are discussed.     

草鱼产卵类型的研究

本文是对已达性成熟年龄的池养和江河雌性草鱼产卵前后卵巢组织学结构的研究。实验结果证明草鱼是一次产卵类型。经人工催产后的雌性草鱼,由于亲鱼的成熟程度存在个体间的差异,有的全产,有的部分产。在湘江天然产卵场捕得的雌性草鱼,也有全产和部分产的。人工催产全产后的卵巢组织学结构是Ⅰ、Ⅱ时相,部分产后的卵巢组织学结构是Ⅰ、Ⅱ和Ⅳ(Ⅳ+、Ⅳ++)时相,已达满熟阶段但未经人工催产的卵巢组织学结构是Ⅰ、Ⅱ和Ⅳ(Ⅳ+++)时相,以上都未发现有处于Ⅲ时相的卵母细胞。从江河天然产卵场捕得的全产、部分产和尚未产卵的雌性草鱼的卵巢组织学结构,与上述结果一致。证实由Ⅲ时相到Ⅳ时相是同步性的。5月全产后的雌性草鱼,其卵巢组织学结构在6-9月内处于第Ⅱ期,没有新的Ⅳ时相卵母细胞。因此,夏季全产后的雌性草鱼,不可能在当年夏季或秋季完成由Ⅱ-Ⅲ-Ⅳ时相的发育程序。草鱼的卵巢成熟系数在繁殖季节只出现一次高峰。       

In vitro developmental competence of buffalo oocytes collected at various stages of the estrous cycle

The objective was to determine the in vitro developmental competence of buffalo oocytes collected from abattoir-derived ovaries at various stages of the estrous cycle and follicular status. In Experiment 1, ovaries (n=476 pairs) were collected and divided into the following five groups: (a) ovaries with a corpus hemorragicum and no dominant follicle (CH-NO-DF); (b) ovaries with a mature functional corpus luteum (CL) and a dominant follicle (CL-DF); (c) ovaries with a mature functional CL and no dominant follicle (CL-NO-DF); (d) ovaries with a regressing CL and a dominant follicle (RCL-DF); and (e) ovaries without any luteal structures and only small follicles (ANEST). In Experiment 2, 144 pairs of ovaries with a CL (or regressing CL) and a dominant follicle were collected and follicles were classified as dominant, largest subordinate, and subordinate. In both experiments, the dominant follicle was defined as any follicle >10mm in diameter that exceeded the diameter of all other (subordinate) follicles. Although oocytes were collected from each group of ovaries, only Grades A or B oocytes were used for in vitro embryo production. Cleavage rates were higher (P<0.05) from oocytes collected from ovaries in the CH-NO-DF (59.6%) and CL-NO-DF (59.2%) groups than those collected from CL-DF (52.2%) and ANEST (43.6%) groups. The yield of transferable embryos was higher (P<0.05) from oocytes collected from CH-NO-DF (27.4%) and CL-NO-DF (24.0%) ovaries than from CL-DF (16.2%), RCL-DF (15.4%), and lowest (P<0.05) from ANEST (8.8%). In Experiment 2, oocytes from the dominant follicle had a higher (P<0.05) cleavage rate (65.2 %) and transferable embryo yield (30.2%) than those collected from the largest subordinate and subordinate follicles. In conclusion, oocyte competence depended on the morphofunctional state of ovaries. Oocyte development was maximal in pairs of ovaries with a corpus hemorragicum or CL and no dominant follicle; in paired ovaries with a CL and a dominant follicle, development was maximal in oocytes derived from the dominant follicle.     

Morphological diversity of oocytes released from the adult mouse ovary.

This study was designed to classify and differentiate the population of oocytes released from the mouse ovary by mechanical means. Liberated oocytes were classified on the basis of their number, size and by the nature of the attachment of follicle cells to these oocytes. Microscopical examination of oocytes mechanically released from the ovaries revealed three distinct morphological classifications of oocytes: (1) those completely devoid of follicle cells, (2) those encased in follicle cells and (3) those in the process of degeneration. Ooxytes ranged in size from approximatley 30mu to 119mu. Those oocytes surrounded by follicle cells could be further subdivided into two groups depending on whether the follicle cells could be mechanically removed from the oocyte. These data demonstrate that the adult ovary contains a variety of classes of oocytes which differ in size and in the extent to which the follicle cells are attached to the oocytes. It is suggested that the metabolic activities and meiotic potnetials of the various oocyte populations differ and as a result, care should be employed to insure a uniform population of oocytes when conducting further studies sith mechanically liberated oocytes.