Oogenesis and spawn formation in the invasive lionfish, Pterois miles and Pterois volitans

The Indo-Pacific lionfish, Pterois miles and P. volitans, have recently invaded the U.S. east coast and the Caribbean and pose a significant threat to native reef fish communities. Few studies have documented reproduction in pteroines from the Indo-Pacific. This study provides a description of oogenesis and spawn formation in P. miles and P. volitans collected from offshore waters of North Carolina, U.S.A and the Bahamas. Using histological and laboratory observations, we found no differences in reproductive biology between P. miles and P. volitans. These lionfish spawn buoyant eggs that are encased in a hollow mass of mucus produced by specialized secretory cells of the ovarian wall complex. Oocytes develop on highly vascularized peduncles with all oocyte stages present in the ovary of spawning females and the most mature oocytes placed terminally, near the ovarian lumen. Given these ovarian characteristics, these lionfish are asynchronous, indeterminate batch spawners and are thus capable of sustained reproduction throughout the year when conditions are suitable. This mode of reproduction could have contributed to the recent and rapid establishment of these lionfish in the northwestern Atlantic and Caribbean.

Zebraflsh z-otu, a novel Otu and Tudor domain-containing gene, is expressed in early stages of oogenesis and embryogenesis

Several studies have suggested that Otu domain had de-ubiquitinating activity and Tudor domain was important for the formation of germ cells. Here, we reported a novel zebrafish ovary-specific gene containing Otu and Tudor domain, z-otu, which was expressed at stages I-III oocytes and embryonic stages from zygotes to early blastula during embryonic cells maintained their totipotency. Therefore, z-otu might link the ubiquitin signaling pathway to early oogenesis and maintaining the totipotency of embryonic cell. (c) 2005 Elsevier B.V. All rights reserved.

Expression pattern and developmental behaviour of cellular nucleic acid-binding protein (CNBP) during folliculogenesis and oogenesis in fish

In vertebrates, folliculogeneis establishes an intricate system for somatic cell-oocyte interaction, and ultimately leads to the acquisition of their respective competences. Although the formation process and corresponding interactions are strikingly similar in diverse organisms, knowledge of genes and signaling pathways involved in follicle formation is very incomplete and the underlying molecular mechanisms remain enigmatic. CNBP has been identified for more than ten years, and the highest level of CNBP transcripts has been observed in adult zebrafish ovary, but little is known about its functional significance during folliculogeneis and oogenesis. In this study, we clone CNBP cDNA from gibel carp (Carassius auratus gibelio), and demonstrate its predominant expression in gibel carp ovary and testis not only by RTPCR but also by Western blot. Its full-length cDNA is 1402 bp, and has an ORF of 489 nt for encoding a peptide of 163 aa. And its complete amino acid sequence shared 68.5%-96.8% identity with CNBPs from other vertebrates. Based on the expression characterization, we further analyze its expression pattern and developmental behaviour during folliculogeneis and oogenesis. Following these studies, we reveal an unexpected discovery that the CagCNBP is associated with follicular cells and oocytes, and significant distribution changes have occurred in degenerating and regenerating follicles. More interestingly, the CagCNBP is more highly expressed in some clusters of interconnected cells within ovarian cysts, no matter whether the cell clusters are formed from the original primordial germ cells or from the newly formed cells from follicular cells that invaded into the atretic oocytes. It is the first time to reveal CNBP relevance to folliculogeneis and oogenesis. Moreover, a similar stage-specific and cell-specific expression pattern has also been observed in the gibel carp testis. Therefore, further studies on CNBP expression pattern and developmental behaviour will be of significance for understanding functional roles of CNBP during gametogenests. (c) 2005 Elsevier B.V. All rights reserved.

Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate

The RNA helicase Vasa is a germ cell marker in animals, and its homolog in vertebrates to date has been limited to bisexual reproduction. We cloned and characterized CagVasa, a Vasa homolog from the gibel carp, a fish that reproduces bisexually or gynogenetically. CagVasa possesses 14 RGG repeats and eight conserved motifs of Vasa proteins. In bisexually reproducing gibel carp, vasa is maternally supplied and its zygotic expression is restricted to gonads. By in situ hybridization on testicular sections, vasa is low in spermatogonia, high in primary spermatocytes, reduced in secondary spermatocytes, but disappears in spermatids and sperm. In contrast, vasa persists throughout oogenesis, displaying low-high-low levels from oogonia over vitellogenic oocytes to maturing oocytes. A rabbit anti-Vasa antibody (alpha Vasa) was raised against the N-terminal CagVasa for fluorescent immunohistochemistry. On testicular sections, Vasa is the highest in spermatogonia, reduced in spermatocytes, low in spermatids, and absent in sperm. In the ovary, Vasa is the highest in oogonia but persists throughout oogenesis. Subcellular localization of vasa and its protein changes dynamically during oogenesis. The aVasa stains putative primordial germ cells in gibel carp fry. It detects gonadal germ cells also in several other teleosts. Therefore, Cagvasa encodes a Vasa ortholog that is differentially expressed in the testis and ovary. Interestingly, the alpha Vasa in combination with a nuclear dye can differentiate critical stages of spermatogenesis and oogenesis in fish. The cross-reactivity and the ability to stain stage-specific germ cells make this antibody a useful tool to identify fish germ cell development and differentiation. (c) 2005 Wiley-Liss, Inc.

Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis

A new gene with WD domains is cloned and characterized according to its differential transcription and expression between previtellogenic oocytes (phase I oocytes) and fully-grown oocytes (phase V oocytes) from natural gynogenetic silver crucian carp (Carassius auratus gibelio) by using the combinative methods of suppressive subtraction hybridization, SMART cDNA synthesis and RACE-PCR. The full-length cDNA is 1870 bp. Its 5 ' untranslated region is 210 bp, followed by an open reading frame of 990 bp, which has the typical vertebrate initiator codon of ANNATG. The open reading frame encodes a protein with 329 amino acids. It has 670 bp of 3 ' untranslated region and an AATAAA polyadenylation signal. Because it has 92% homology to STRAP (serine-threonine kinase receptor-associated protein), a recently reported gene, we named it FSTRAP (fish STRAP). Virtual Northern blotting indicated that the FSTRAP was transcribed in fully-grown oocytes (phase V oocytes), but not in previtellogenic oocytes (phase I oocytes). RT-PCR analysis showed that FSTRAP was transcribed in brain, heart, kidney, muscle, ovary, spleen and testis, but not in liver. And its mRNA could be detected in the oocytes from phase II to phase V. Western blotting also showed that FSTRAP protein could be detected in brain, heart, kidney, muscle, ovary, spleen and testis except liver. Results of Western blotting on various oocytes were also similar to the RT-PCR data. FSTRAP protein was not expressed in the previtellogenic oocytes. Its expression initiated from phase II oocytes after vitellogenesis, and was consistent with the mRNA transcription.

Centriole elimination in Drosophila melanogaster oogenesis

Dissertation presented to obtain the Ph.D degree in Cellular Biology

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of X (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data. suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens. (c) 2009 Elsevier Ltd. All rights reserved.

Ultrastructural aspects of oogenesis and oocyte primary growth in Serrasalmus spilopleura (Teleostei, Characiformes, Characidae)

The ultrastructural characteristics of the organelles present in Serrasalmus spilopleura oogonia and oocytes undergoing primary growth were described in detail, considering its role in the nuclear and cytoplasmic metabolic processes that occur in these cell types. Even though these cells do not significantly differ from those similar to them that are found in other teleost groups, the analysis of their ultrastructure makes available new data on the reproductive biology of Characiformes. (C) 2001 Harcourt Publishers Ltd.

Occurrence and ultrastructural characterization of nuage during oogenesis and early spermatogenesis of Piaractus mesopotamicus Holmberg, 1887 (Teleostei)

Foi investigada a ocorrência e caracterizada ultra-estruturalmente a eliminação de material nuclear eletrondenso (nuage) para o citoplasma durante a ovogênese e durante os estágios iniciais da espermatogênese de Piaractus mesopotamicus, um peixe do Pantanal Matogrossense de ciclo reprodutivo sazonal. Constataram-se nas células germinativas femininas dois momentos de eliminação desse material, na ovogônia e no ovócito em fase perinucleolar. Nas células masculinas, material com morfologia e comportamento muito semelhante foi encontrado na espermatogônia. em todos os casos, o material associou-se a mitocôndrias. A possível função desse material foi discutida.

Ovarian morphology and oogenesis dynamic of the diplopod Urostreptus atrobrunneus Pierozzi & Fontanetti, 2006 (Spirostreptidae), a potential plague in urban centers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytoskeletal organization of bee ovarian follicles during oogenesis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dynamics and cytochemistry of oogenesis in Astyanax fasciatus (Cuvier) (Teleostei, Characiformes, Characidae) from Rio Sapucaí, Minas Gerais State, Brazil

Oogenesis involves a set of transformations which are undergone by female germ cells These cells change into oogonias and then into mature oocytes. Sexually mature female fish were collected monthly, during one year, from the Sapucaí River, a tributary of the Rio Grande, which is part of the Furnas Reservoir in the state of Minas Gerais. During the several stages of maturation, we observed small round oogonias with a large nucleus, a single nucleolus, and weakly stained cytoplasm with eosinophilic granules. The primary oocytes showed a large basophilic nucleus, with a developed peripheral nucleolus and a reduced cytoplasm. The previtellogenic oocytes presented voluminous cytoplasm and nucleus with several small peripheral nucleoli. The oocytes underwent vitellogenesis with the development of the zona radiata and the follicle cells. Their cytochemical reactions indicated that the two layers of the zona radiata of A. fasciatus contained proteins and polysaccharides. The initially squamous follicle cells, became cuboidal. In mature oocytes, the nucleus moved toward the periphery, next to the micropyle, and the yolk granules formed by proteins, fulfilled the cytoplasm. The clear unstained vesicles are likely to be the cortical alveoli in the perivitelline region.

Dynamics and cytochemistry of oogenesis in Leporinus striatus Kner (Teleostei, Characiformes, Anostomidae) from the Rio Sapucaí, Minas Gerais State, Brazil

Oogenesis involves a sequense of transformations which are undergone by female germ cells. These cells change into oogonias and then into mature oocytes. Sexually mature females were collected monthly, during one year, from the Rio Sapucaí, tributary of the Rio Grande, which is part of the Furnas Reservoir system in the state of Minas Gerais. The observed material showed that oogonias were small spherical cells, had a big spherical nucleus, with a single nucleolus, and weakly stained cytoplasm with eosinophilic granules (FG stained), which indicate their protein content. The primary oocytes showed a big basophilic nucleus, with a large peripheral nucleolus, and several smaller nucleoli. They show a reduced cytoplasmic content. The previtellogenic oocytes presented voluminous cytoplasm and nucleus with several small peripheral nucleoli. The oocytes underwent vitellogenesis with the development of the zona radiata and the follicle cells. The zona radiata had two layers, the outer and the inner, which showed its protein content when stained with CM and FG techniques. TB pH 2.5 and pH 4.0 staining showed that oocytes undergoing vitellogenesis presented weakly stained cytoplasm and peripheral cytoplasmic vesicles. The follicle cells that were squamous became cuboidal. In mature oocytes, the yolk granules that filled the cytoplasm became green and blue when stained with FG and CM techniques, indicating their protein content. The perivitclline region showed rosy stained vesicles (TB pH 2.5 and pH 4.0) spread among the weakly stained peripheral vesicles, which seemed to be the cortical alveoli. The zona radiata cells, CM and FG stained, still showed two layers like the oocytes from the previous stage, but thicker.