Life stages and reproductive components of the Marmorkrebs (marbled crayfish), the first parthenogenetic decapod crustacean

Recently, we briefly reported on the first case of parthenogenesis in the decapod Crustacea which was found in the Marmorkrebs or marbled crayfish, a cambarid species of unknown geographic origin and species identity. Curiously, this animal is known only from aquarium populations, where it explosively propagates. By means of light and electron microscopic techniques we have now investigated the reproductive components of this crayfish, using more than 100 specimens ranging from hatchling to repeatedly spawned adult. Additionally, we documented its principal life stages. Our results revealed that the external sexual characters and also the gonads of the marbled crayfish are purely female, making this fast-reproducing species a good model for investigating female reproductive features in crayfish. Testicular tissues, ovotestes, or male gonoducts, gonopores, or gonopods were never found, either in small juveniles or large adult specimens, confirming the parthenogenetic nature of this crayfish. Parthenogenesis may have arisen spontaneously or by interspecific hybridization since Wolbachia-like feminizing microorganisms were not found in the ovaries. The external sexual characters of the marbled crayfish are first recognized in Stage 4 juveniles and are structurally complete approximately 2 months after hatching in specimens of approximately 2 cm total length. In the same life stage the ovary is fully differentiated as well, although the oocytes are in previtellogenic and primary vitellogenic stages only. The architecture of the mature ovary and also the synchronous maturation of cohorts of primary vitellogenic oocytes by secondary vitellogenesis are in general agreement with data published on ovaries of bisexual crayfish. New results were obtained with respect to the muscular nature of the ovarian envelope and its extensive proliferation after the first spawning, the distribution of hemal sinuses in the ovarian envelope and in the interstitium around the oogenetic pouches, the high transport activity of the follicle cells, and the colonization of oogenetic pouches by previtellogenic oocytes that originate in the germaria. Investigation of the nuclei of oocytes in the germaria and oogenetic pouches revealed no signs of meiosis, as usually found in females of bisexual decapods, suggesting that parthenogenesis in the marbled crayfish might be an apomictic thelytoky. The detection of new rickettsial and coccidian infections in the ovary and further organs raises fears that the marbled crayfish might endanger native European species by transmission of pathogens once escaped into the wild.     

Previtellogenic oocytes of South African largemouth bass Micropterus salmoides Lacépède 1802 (Actinopterygii,Perciformes) - the Balbiani body,cortical alveoli and developing eggshell

The ovaries of the largemouth bass Micropterus salmoides, an alien and invasive species in South Africa, contain a germinal epithelium which consists of germline and somatic cells, as well as previtellogenic and late vitellogenic ovarian follicles. The ovarian follicle consists of an oocyte surrounded by follicular cells and a basal lamina; thecal cells adjacent to this lamina are covered by an extracellular matrix. In this article, we describe the Balbiani body and the polarization and ultrastructure of the cytoplasm (ooplasm) in previtellogenic oocytes. The nucleoplasm in all examined oocytes contains lampbrush chromosomes, nuclear bodies and several nucleoli near the nuclear envelope. The ultrastructure of the nucleoli is described. Numerous nuage aggregations are present in the perinuclear cytoplasm in germline cells as well as in the ooplasm. Possible roles of these aggregations are discussed. The ooplasm contains the Balbiani body, which defines the future vegetal region in early previtellogenic oocytes. It is comprised of nuage aggregations, rough endoplasmic reticulum, Golgi apparatus, mitochondria, complexes of mitochondria with nuage-like material, and lysosome-like organelles. In mid-previtellogenic oocytes, the Balbiani body surrounds the nucleus and later disperses in the ooplasm. The lysosome-like organelles fuse and transform into vesicles containing material which is highly electron dense. As a result of the fusion of the vesicles of Golgi and rough endoplasmic reticulum, the cortical alveoli arise and distribute uniformly throughout the ooplasm of late previtellogenic oocytes. During this stage, the deposition of the eggshell (zona radiata) begins. The eggshell is penetrated by canals containing microvilli and consists of the following: the internal and the external egg envelope. In the external envelope three sublayers can be distinguished.     

Possible participation of mitochondria in lipid yolk formation in oocytes of paddlefish and sturgeon

The ovary of paddlefish and sturgeons (Acipenseriformes) is composed of discrete units: the ovarian nests and ovarian follicles. The ovarian nests comprise oogonia and numerous early dictyotene oocytes surrounded by somatic prefollicular cells. Each ovarian follicle consists of a spherical oocyte and a layer of follicular cells situated on a thick basal lamina, encompassed by thecal cells. The cytoplasm of previtellogenic oocytes is differentiated into two distinct zones: the homogeneous and granular zones. The homogeneous cytoplasm is organelle-free, whereas the granular cytoplasm contains numerous organelles, including mitochondria and lipid droplets. We have analyzed the cytoplasm of early dictyotene and previtellogenic oocytes ultrastructurally and histologically. In the cytoplasm of early dictyotene oocytes, two morphologically different types of mitochondria can be distinguished: (1) with well-developed cristae and (2) with distorted and fused cristae. In previtellogenic oocytes, the mitochondria of the second type show various stages of cristae distortion; they contain and release material morphologically similar to that of lipid droplets and eventually degenerate. This process of mitochondrial transformation is accompanied by an accumulation of lipid droplets that form a single large accumulation (lipid body) located in the vicinity of the oocyte nucleus (germinal vesicle). The lipid body eventually disperses in the oocyte center. The possible participation of these mitochondria in the formation of oocyte lipid droplets is discussed. This work was supported by funds from the research grant BW/IZ/2005 to M.Ż. An erratum to this article can be found at http://dx.doi.org/. An erratum to this article can be found at     

Ultrastructural observations of previtellogenic ovarian follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii

The ultrastructural organization of the previtellogenic follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii, of the Western Ghats of India, were observed. Both species follow a similar seasonal reproductive pattern. The ovaries contain primordial follicles throughout the year with previtellogenic, vitellogenic, or postvitellogenic follicles, depending upon the reproductive status. The just-recruited primordial follicle includes an oocyte surrounded by a single layer of follicle and thecal cells. The differentiation of the theca into externa and interna layers, the follicle cells into dark and light variants, and the appearance of primordial yolk platelets and mitochondrial clouds in the ooplasm mark the transition of the primordial follicle into a previtellogenic follicle. During further development of the previtellogenic follicle the following changes occur: i) the theca loses distinction as externa and interna; ii) all the follicle cells become the dark variant and increase in the complexity of ultrastructural organization; iii) the nucleus of the oocyte transforms into the germinal vesicle and there is amplification of the nucleoli; iv) the primordial yolk platelets of the cortical cytoplasm of the oocyte increase in abundance; v) the mitochondrial clouds fragment and the mitochondria move away from the clouds, leaving behind the cementing matrix, which contains membrane-bound vesicles of various sizes, either empty or filled with a lipid material; vi) the perivitelline space appears first as troughs at the junctional points between the follicle cells and oocyte, which subsequently spread all around to become continuous; vii) macrovilli and microvilli develop from the follicle cells and oocyte, respectively; and viii) the precursor material of the vitelline envelop arrives at the perivitelline space. The sequential changes in the previtellogenic follicles of two species of caecilians are described.     

Ovarian germinal epithelium and folliculogenesis in the common snook, Centropomus undecimalis (Teleostei: centropomidae)

The ovarian germinal epithelium in the common snook, Centropomus undecimalis, is described. It consists of epithelial and prefollicle cells that surround germ cells, either oogonia or oocytes, respectively. The germinal epithelium borders a body cavity, the ovarian lumen, and is supported by a basement membrane that also separates the epithelial compartment of the ovarian lamellae from the stromal compartment. During folliculogenesis, the epithelial cells, whose cytoplasmic processes encompass meiotic oocytes, transform into prefollicle cells, which become follicle cells at the completion of folliculogenesis. The follicle is a derivative of the germinal epithelium and is composed of the oocyte and surrounding follicle cells. It is separated from the encompassing theca by a basement membrane. The cells that form the theca interna are derived from prethecal cells within the extravascular space of the ovarian stroma. The theca externa differentiates from undifferentiated cells within the stromal compartment of the ovary, from within the extravascular space. The theca interna and the theca externa are not considered to be part of the follicle and are derived from a different ovarian compartment than the follicle. Meiosis commences while oocytes are still within the germinal epithelium and proceeds as far as arrested diplotene of the first meiotic prophase. The primary growth phase of oocyte development also begins while oocytes are still within the germinal epithelium or attached to it in a cell nest. The definitions used herein are consistent between sexes and with the mammalian literature.     

Involvement of the gonadal germinal epithelium during sex reversal and seasonal testicular cycling in the protogynous swamp eel,Synbranchus marmoratus Bloch 1795 (Teleostei,Synbranchidae)

The swamp eel, Synbranchus marmoratus, is a protogynous, diandric species. During sex reversal, the ovarian germinal epithelium, which forms follicles containing an oocyte and encompassing follicle cells during the female portion of the life cycle, produces numerous invaginations, or acini, into the ovarian stroma. Within the acini, the gonia that formerly produced oocytes become spermatogonia, enter meiosis, and produce sperm. The acini are bounded by the basement membrane of the germinal epithelium. Epithelial cells of the female germinal epithelium, which formerly became follicle (granulosa) cells, now become Sertoli cells in the developing testis. Subsequently, lobules and testicular ducts form. The swamp eel testis has a lobular germinal compartment in both primary and secondary males, although the germinal compartment in testes of secondary males resides within the former ovarian lamellae. The germinal compartment, supported by a basement membrane, is composed of Sertoli and germ cells that give rise to sperm. Histological and immunohistochemical techniques were used to describe the five reproductive classes that were observed to occur during the annual reproductive cycle: regressed, early maturation, mid-maturation, late maturation, and regression. These classes are differentiated by the presence of continuous or discontinuous germinal epithelia and by the types of germ cells present. Synbranchus marmoratus has a permanent germinal epithelium. Differences between the germinal compartment of the testes of primary and secondary males were not observed.     

Protandric hermaphrodite peculiarities in Amphiprion frenatus Brevoort (Teleostei, Pomacentridae)

Electron microscopy of the male phase of the ovotestis of Amphiprion frenatus , a protandric hermaphrodite, showed no connective tissue between male and female areas and, as the basal lamina was lacking both along the seminiferous tubules and round the previtellogenic oocytes, the male and female germ cells were only separated by their respective surrounding somatic cells (Sertoli and follicle cells). Besides previtellogenic oocytes, oocytes in meiotic prophase and very small (young) previtellogenic oocytes, were detected in the ovarian part, as spermatogenesis proceeded, revealing oogenetic activity. Degeneration of some previtellogenic oocytes and their follicle cells was discernible.     

Identification of cells migrating from the thecal layer of ovarian follicles

In the mammalian ovary, oocytes are contained within ovarian follicles. These consist in an oocyte surrounded by supporting cells: an inner layer of granulosa cells and an outer layer of thecal cells separated by a basal lamina. At any one time, a developing cohort of follicles exists, from which only a small species-specific number are selected for continued development towards ovulation, with the remainder dying by follicular atresia. Here, we use in vitro methods to study interactions between two follicles in culture (follicle co-cultures). We show that, when two individual follicles are grown together in culture, cells and cellular processes migrate from the outer thecal layer of one follicle to the thecal layer of the other co-cultured follicle. These cells are identified as a mixed population containing primarily endothelial but also neuronal cells. Both are able to migrate through the ovarian interstitum, making contact with the basal lamina of other follicles and with similar cells from these other follicles. Networks of such cells might be involved in interfollicular communication and in the coordination of follicle selection for ovulation.     

Ovary of the pipefish,Syngnathus scovelli

Gross dissection, light microscopy, and transmission electron microscopy were used to generate a detailed understanding of the ovarian anatomy of the pipefish, Syngnathus scovelli. The ovary is a cylindrical tube bounded by an outer layer consisting of a smooth muscle wall and an inner layer of luminal epithelium, with follicles sandwiched between the two layers. A remarkable feature of this ovary is a sequential pattern of follicle development. This pattern begins at the germinal ridge with a gradient of follicles of increasing developmental age extending to the mature edge. The germinal ridge is an outpocketed region of the luminal epithelium containing early germinal cells and somatic prefollicular cells. Therefore, the germinal ridge and luminal epithelium share the same ovarian compartment and follicle formation occurs within this compartment. The mature edge is defined as the site of oocyte maturation and ovulation. The outer ovarian wall contains unmyelinated nerve fibers throughout. Longitudinally oriented unmyelinated nerves are also observed near the smooth muscle bundles associated with the mature edge. Oocytes near the mature edge are polarized such that the germinal vesicle (nucleus) is generally oriented toward the luminal epithelium. The sandwichlike organization of the ovary results in follicles that have a shared theca. An extensive lymphatic network is also interspersed among the follicles. Thus, the exceptional features of the pipefish ovary make it particularly well suited for the examination of early events in oogenesis. Specifically, we characterize pipefish folliculogenesis in detail.     

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.     

Cytokeratin cytoskeleton in the differentiating ovarian follicle of the lizard Podarcis sicula Raf

By immunoblotting and immunocytochemical techniques, we characterized the cytokeratins previously localized by us in the previtellogenic ovarian follicle of Podarcis sicula. Our results show that these cytokeratins correspond to those expressed in the monolayered epithelia. In fact, the immunoblotting analysis showed that the NCL-5D3 antibody, specific for human low molecular weight cytokeratins expressed in monolayered epithelia, reacted with the cytokeratins extracted both from the ovary and from the monolayered intestinal mucosa of Podarcis sicula. Furthermore, this antibody, in this reptile as in humans, clearly immunolabeled sections of corresponding tissues. The organization of the cytokeratin cytoskeleton in the main steps of the ovarian follicle differentiation was also clarified. The reported observations suggest that in Podarcis sicula, the cytokeratin cytoskeleton is absent in the early oocytes. It first appears in the growing oocytes as a thin cortical layer in concomitance with its becoming visible also in the enlarging follicle cells. In the larger follicles, this cytoskeleton appears well organized in intermediate cells and in particular in fully differentiated pyriform cells. In both these cells a cytokeratin network connects the cytoplasm to the oocyte cortex through intercellular bridges. At the end of the previtellogenic oocyte growth, the intense immunolabeling of the apex in the regressing pyriform cells suggests that the cytokeratin, as other cytoplasmic components, may be transferred from these follicle cells to the oocyte. At the end of the oocyte growth, in the larger vitellogenic oocytes surrounded by a monolayer of follicle cells, the cytokeratin constitutes a heavily immunolabeled cortical layer thicker than in the previous stages. Mol. Reprod. Dev. 48:536–542, 1997. © 1997 Wiley-Liss, Inc.     

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.     

Ovary of the seahorse,Hippocampus erectus

The ovary of the seahorse, Hippocampus erectus, is a cylindrical tube bounded by an outer layer consisting of a mesothelium and muscular wall and by an inner luminal epithelium, with a single row of developing follicles sandwiched between the two layers. Follicles are produced by a germinal ridge, which contains oogonia, early oocytes, and prefollicle cells, and which runs along the length of the ovary. The germinal ridge is an outpocketing of the luminal epithelium, as indicated by a continuous underlying basal lamina. Prefollicle cells invest diplotene oocytes and the complex eventually pinches off the germinal ridge as a primordial follicle surrounded by a basal lamina derived from the germinal ridge. Subsequent investment of the primordial follicle by elements of the theca complete the process of folliculogenesis. H. erectus has two ovaries and each ovary has two dorsally located germinal ridges. Thus, in each ovary the derived follicular lamina is bilaterally symmetrical: two temporally and spatially arranged sequences of developing follicles are produced, with the largest follicles found along the ventral midline of the ovary. The advantages of developmental, kinetic, and systemic analyses of these unusual ovaries are indicated.     

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.     

The localization of CD3, CD79a,CD68 and S100 protein immunoreactive cells in hemal nodes of Saanen goat (Capra hircus)

We investigated the structure of hemal nodes in Saanen goats using immunohistochemical staining. We examined the distribution of CD3 positive T lymphocytes, CD79a positive B lymphocytes, CD68 positive macrophages and S100 protein positive follicular dendritic cells. Hemal nodes of six healty adult female goats were used. Hemal nodes were removed from the thoracic and abdominal cavities. The oval to round hemal nodes were observed especially between the abdominal aorta and vena cava, and near the kidneys and adrenal glands. Tissue sections were stained with Crossmon’s modified triple stain to demonstrate general histological structure. The avidin-biotin-peroxidase technique using anti-CD3, anti-CD79a, anti-CD68 and anti-S100 primary antibodies was used for immunohistochemistry. Many CD3 positive T lymphocytes were found in the germinal center of the lymph follicles and in the lymphatic cords of hemal nodes; CD3 positive cells also were observed in the sinuses. CD79a and CD68 positive cells were found at the germinal center of the lymph follicles. In the lymph follicles near the subcapsular sinuses, CD79a and CD68 positive cells were found especially in e areas bordering the mantle zone. S100 positive cells were found in the lymph follicles, lymphatic cords and sinuses.     

Structure of ovaries and oogenesis in Corydalidae and Chauliodidae (Insecta, Megaloptera). I. Architecture of adult ovarioles and previtellogenesis.

The results of histological and EM studies on the ovaries of three representatives of Megaloptera: Chauliodes pectinicornis, Nigronia fasciata (Chauliodidae), and Corydalus peruvianus Corydalidae) are presented. It is shown that the ovaries of all 3 investigated species are panoistic (secondary panoistic, = neopanoistic) and consist of numerous (more than a hundred) ovarioles that are differentiated into 3 well-defined regions: the terminal filament, the germarium, and the vitellarium. The germaria of adult females are apparently non-functional and contain germ and somatic cells in various stages of degeneration. The vitellaria are composed of 12-15 developing ovarian follicles (= oocytes surrounded by follicular cells) in a linear arrangement. In adult females these follicles can be classified into early previtellogenic, late previtellogenic, vitellogenic, and choriogenic. During early previtellogenesis oocyte nuclei (= germinal vesicles) contain single nucleolar masses. Histochemical analyses indicate that within the masses DNA as well as AgNOR proteins are present. During subsequent stages of the previtellogenic growth nucleolar masses gradually break down into smaller aggregations of coarse granular material, i.e. multiple nucleoli. In chauliodids the nucleoli are distributed evenly throughout the nucleoplasm while in the corydalid, C. peruvianus, they form a characteristic ring. The presented results are discussed in a phylogenetic context.     

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.     

Phylogenetic significance of the structure of adult ovary and oogenesis in a primitive chilognathan diplopod,Hyleoglomeris japonica Verhoeff (Glomerida,Diplopoda)

Some histological details of the adult ovary of Hyleoglomeris japonica are described for the first time in the glomerid diplopods. The ovary is a single, long sac-like organ extending from the 4th to the 12th body segment along the median body axis, lying between the alimentary canal and the ventral nerve cord. The ovarian wall consists of a layer of thin ovarian epithelium which surrounds a wide ovarian lumen. A pair of longitudinal “germ zones,” including female germ cells, runs in the lateral ovarian wall. Each germ zone consists of two types of oogenetic areas: 1) 8–12 narrow patch-shaped areas for oogonial proliferation, arranged metamerically in a row along each of the dorsal and ventral peripheries, and 2) the remaining wide area for oocyte growth. Oogonial proliferation areas include oogonia, very early previtellogenic oocytes, and young somatic interstitial cells, among the ovarian epithelial cells. The larger early previtellogenic oocytes in the oogonial proliferation areas are located nearer to the oocyte growth area, and migrate to the oocyte growth area. They are surrounded by a layer of follicle cells and are connected with the ovarian epithelium of the oocyte growth area by a portion of their follicles. They grow into the ovarian lumen, but their follicles are still connected with the oocyte growth area. Various sizes of the previtellogenic and vitellogenic oocytes in the ovarian lumen are connected with the oocyte growth area; the smaller oocytes are connected nearer to the dorsal and ventral oogonial proliferation areas, while the larger ones are connected nearer to the longitudinal middle line of the oocyte growth area. Following the completion of vitellogenesis and egg membrane formation in the largest primary oocytes, the germinal vesicles break down. Ripe oocytes are released from their follicles directly into the ovarian lumen to be transported into the oviducts. Ovarian structure and oogenesis of H. japonica are very similar to those of other chilognathan diplopods. At the same time, however, some characteristic features of the ovary of H. japonica are helpful for understanding the structure and evolution of the diplopod ovaries. Some aspects of the phylogenetic significance in the paired germ zones of H. japonica are discussed. J. Morphol 231:277–285, 1997. © 1997 Wiley-Liss, Inc.     

Ultrastructure of the ovarian follicles in the placentotrophic Andean lizard of the genus Mabuya (Squamata: Scincidae)

We studied the ultrastructural organization of the ovarian follicles in a placentotrophic Andean lizard of the genus Mabuya. The oocyte of the primary follicle is surrounded by a single layer of follicle cells. During the previtellogenic stages, these cells become stratified and differentiated in three cell types: small, intermediate, and large globoid, non pyriform cells. Fluid‐filled spaces arise among follicular cells in late previtellogenic follicles and provide evidence of cell lysis. In vitellogenic follicles, the follicular cells constitute a monolayered granulosa with large lacunar spaces; the content of their cytoplasm is released to the perivitelline space where the zona pellucida is formed. The oolemma of younger oocytes presents incipient short projections; as the oocyte grows, these projections become organized in a microvillar surface. During vitellogenesis, cannaliculi develop from the base of the microvilli and internalize materials by endocytosis. In the juxtanuclear ooplasm of early previtellogenic follicles, the Balbiani's vitelline body is found as an aggregate of organelles and lipid droplets; this complex of organelles disperses in the ooplasm during oocyte growth. In late previtellogenesis, membranous organelles are especially abundant in the peripheral ooplasm, whereas abundant vesicles and granular material occur in the medullar ooplasm. The ooplasm of vitellogenic follicles shows a peripheral band constituted by abundant membranous organelles and numerous vesicular bodies, some of them with a small lipoprotein core. No organized yolk platelets, like in lecithotrophic reptiles, were observed. Toward the medullary ooplasm, electron‐lucent vesicles become larger in size containing remains of cytoplasmic material in dissolution. The results of this study demonstrate structural similarities between the follicles of this species and other Squamata; however, the ooplasm of the mature oocyte of Mabuya is morphologically similar to the ooplasm of mature oocytes of marsupials, suggesting an interesting evolutionary convergence related to the evolution of placentotrophy and of microlecithal eggs. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Histological and enzyme histochemical investigation of the hemal nodes of the hair goat

We investigated the structure of the hemal node in six healthy hair goats using histological and enzyme histochemical methods. After processing, tissue sections were stained with Crossman's trichrome, Gordon-Sweet's silver and Pappenheim's panoptic stains. Alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) were demonstrated in frozen sections. Hemal nodes were encapsulated by connective tissue and few smooth muscle cells. Several trabeculae originated from the capsule and extended into the hemal node. A subcapsular sinus was present beneath the capsule and was continuous with the deeper sinuses. Subcapsular and deep sinuses were filled with erythrocytes. The parenchyma consisted of lymphoid follicles, diffuse interfollicular lymphocytes and irregular wide lymphoid cords. Cortical and medullary regions were not distinct. ANAE (+) and ACP-ase (+) cells were located mainly in the germinal centers of the lymphoid follicles and also were scattered equally in the interfollicular region and lymphoid cords. Monocytes, macrophages and reticular cells displayed a diffuse positive reaction, whereas localized granular positivity was observed in lymphocytes. We demonstrated that the general structure of the hair goat hemal nodes is similar to that of other ruminant species.