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Cell and Tissue Research Oct 2016Oocytes of many invertebrate and vertebrate species contain a characteristic organelle complex known as the Balbiani body (Bb). Until now, three principal functions have...
Oocytes of many invertebrate and vertebrate species contain a characteristic organelle complex known as the Balbiani body (Bb). Until now, three principal functions have been ascribed to this complex: delivery of germ cell determinants and localized RNAs to the vegetal cortex/posterior pole of the oocyte, transport of the mitochondria towards the germ plasm, and participation in the formation of lipid droplets. Here, we present the results of a computer-aided 3D reconstruction of the Bb in the growing oocytes of an insect, Thermobia domestica. Our analyses have shown that, in Thermobia, the central part of each fully developed Bb comprises a single intricate mitochondrial network. This "core" network is surrounded by several isolated bean-shaped mitochondrial units that display lowered membrane potential and clear signs of degeneration. In light of the above results and recent theoretical models of mitochondrial quality control, the role of the Bb is discussed. We suggest that, in addition to the aforementioned functions, the Bb is implicated in the selective elimination of dysfunctional mitochondria during oogenesis.
Topics: Animals; Apoptosis; Cell Differentiation; Chromosomal Puffs; Imaging, Three-Dimensional; Insecta; Mitochondria; Morphogenesis; Oocytes; Oogenesis
PubMed: 27164893
DOI: 10.1007/s00441-016-2414-x -
Protoplasma Sep 2019It is a first report on the structure of germline cells in ovaries of albino sterlet Acipenser ruthenus L. 1758. Ovarian nests, follicles, and germinal epithelium have...
It is a first report on the structure of germline cells in ovaries of albino sterlet Acipenser ruthenus L. 1758. Ovarian nests, follicles, and germinal epithelium have been examined in gynogenetic and control specimens of this species. The structure of oogonia (named the cystoblasts) and of germline cysts in the nests has been described in detail. Also, the asymmetry in the cytoplasm and early growth of cystocytes in the cysts and of early previtellogenic oocytes has been described. In the cytoplasm of cystoblasts and in all cystocytes, a precursor of granular cytoplasm (Balbiani cytoplasm) is present and defines future vegetal region in the oocytes. Interestingly, the nuclei in cystoblasts comprise a large dense body that contains deoxyribonucleic acid (DNA). The role of this body in formation of multiple nucleoli has been explained. During the zygotene and pachytene stages, massive extrachromosomal amplification of DNA begins in the nucleoplasm of all cystocytes. As a result of the accumulation of extra DNA, an irregularly shaped DNA-body is formed. Multiple nucleoli arise in this DNA-body and around fragments of dense bodies. The asymmetry of the early previtellogenic oocyte cytoplasm is well marked by the presence of the granular cytoplasm. Moreover, the cisternae of the rough endoplasmic reticulum, dictyosomes, mitochondria, complexes of mitochondria with cement, nuage accumulations, and lipid droplets are located in specific zones in the granular cytoplasm. The follicular epithelium is composed of two subpopulations of somatic follicular cells (FCs): the main body cells and future micropylar cells.
Topics: Animals; Cysts; Female; Fishes; Germ Cells; Ovarian Follicle
PubMed: 31020396
DOI: 10.1007/s00709-019-01376-0 -
International Journal of Biological... 2014Germ cells produce sperm and eggs for reproduction and fertility. The Asian seabass (Lates calcarifer), a protandrous marine fish, undergoes male-female sex reversal and...
Germ cells produce sperm and eggs for reproduction and fertility. The Asian seabass (Lates calcarifer), a protandrous marine fish, undergoes male-female sex reversal and thus offers an excellent model to study the role of germ cells in sex differentiation and sex reversal. Here we report the cloning and expression of vasa as a first germ cell marker in this organism. A 2241-bp cDNA was cloned by PCR using degenerate primers of conserved sequences and gene-specific primers. This cDNA contains a polyadenylation signal and a full open reading frame for 645 amino acid residues, which was designated as Lcvasa for the seabass vasa, as its predicted protein is homologous to Vasa proteins. The Lcvasa RNA is maternally supplied and specific to gonads in adulthood. By chromogenic and fluorescent in situ hybridization we revealed germ cell-specific Lcvasa expression in both the testis and ovary. Importantly, Lcvasa shows dynamic patterns of temporospatial expression and subcellular distribution during gametogenesis. At different stages of oogenesis, for example, Lcvasa undergoes nuclear-cytoplasmic redistribution and becomes concentrated preferentially in the Balbiani body of stage-II~III oocytes. Thus, the vasa RNA identifies both female and male germ cells in the Asian seabass, and its expression and distribution delineate critical stages of gametogenesis.
Topics: Amino Acid Sequence; Animals; Base Sequence; Fish Proteins; Fishes; Germ Cells; Male; Molecular Sequence Data; Oocytes; Oogenesis; Phylogeny; Sequence Alignment; Sex Determination Processes; Testis
PubMed: 24550690
DOI: 10.7150/ijbs.6797 -
Science (New York, N.Y.) Apr 2016Oocytes differentiate in diverse species by receiving organelles and cytoplasm from sister germ cells while joined in germline cysts or syncytia. Mouse primordial germ...
Oocytes differentiate in diverse species by receiving organelles and cytoplasm from sister germ cells while joined in germline cysts or syncytia. Mouse primordial germ cells form germline cysts, but the role of cysts in oogenesis is unknown. We find that mouse germ cells receive organelles from neighboring cyst cells and build a Balbiani body to become oocytes, whereas nurselike germ cells die. Organelle movement, Balbiani body formation, and oocyte fate determination are selectively blocked by low levels of microtubule-dependent transport inhibitors. Membrane breakdown within the cyst and an apoptosis-like process are associated with organelle transfer into the oocyte, events reminiscent of nurse cell dumping in Drosophila We propose that cytoplasmic and organelle transport plays an evolutionarily conserved and functionally important role in mammalian oocyte differentiation.
Topics: Animals; Apoptosis; Biological Evolution; Cytoplasm; Female; Giant Cells; Mice; Microtubules; Oocytes; Oogenesis; Organelles
PubMed: 26917595
DOI: 10.1126/science.aad2156 -
Developmental Cell Aug 2018Phase separation represents an important form of subcellular compartmentalization. However, relatively little is known about how the formation or disassembly of such...
Phase separation represents an important form of subcellular compartmentalization. However, relatively little is known about how the formation or disassembly of such compartments is regulated. In zebrafish, the Balbiani body (Bb) and the germ plasm (Gp) are intimately linked phase-separated structures essential for germ cell specification and home to many germ cell-specific mRNAs and proteins. Throughout development, these structures occur as a single large aggregate (Bb), which disperses throughout oogenesis and upon fertilization accumulates again into relatively large assemblies (Gp). Formation of the Bb requires Bucky ball (Buc), a protein with prion-like properties. We found that the multi-tudor domain-containing protein Tdrd6a interacts with Buc, affecting its mobility and aggregation properties. Importantly, lack of this regulatory interaction leads to significant defects in germ cell development. Our work presents insights into how prion-like protein aggregations can be regulated and highlights the biological relevance of such regulatory events.
Topics: Animals; Cytoplasm; Germ Cells; Oocytes; Oogenesis; Organelles; RNA, Messenger; Zebrafish; Zebrafish Proteins
PubMed: 30086300
DOI: 10.1016/j.devcel.2018.07.009 -
Biochemistry Nov 2021In immature oocytes, Balbiani bodies are conserved membraneless condensates implicated in oocyte polarization, the organization of mitochondria, and long-term organelle...
In immature oocytes, Balbiani bodies are conserved membraneless condensates implicated in oocyte polarization, the organization of mitochondria, and long-term organelle and RNA storage. In , Balbiani body assembly is mediated by the protein Velo1. Velo1 contains an N-terminal prion-like domain (PLD) that is essential for Balbiani body formation. PLDs have emerged as a class of intrinsically disordered regions that can undergo various different types of intracellular phase transitions and are often associated with dynamic, liquid-like condensates. Intriguingly, the Velo1 PLD forms solid-like assemblies. Here we sought to understand why Velo1 phase behavior appears to be biophysically distinct from that of other PLD-containing proteins. Through bioinformatic analysis and coarse-grained simulations, we predict that the clustering of aromatic residues and the amino acid composition of residues between aromatics can influence condensate material properties, organization, and the driving forces for assembly. To test our predictions, we redesigned the Velo1 PLD to test the impact of targeted sequence changes We found that the Velo1 design with evenly spaced aromatic residues shows rapid internal dynamics, as probed by fluorescent recovery after photobleaching, even when recruited into Balbiani bodies. Our results suggest that Velo1 might have been selected in evolution for distinctly clustered aromatic residues to maintain the structure of Balbiani bodies in long-lived oocytes. In general, our work identifies several tunable parameters that can be used to augment the condensate material state, offering a road map for the design of synthetic condensates.
Topics: Amino Acids, Aromatic; Animals; Biomolecular Condensates; Cell Polarity; Cells, Cultured; Female; Intravital Microscopy; Oocytes; Phase Transition; Primary Cell Culture; Protein Domains; Protein Engineering; T-Box Domain Proteins; Xenopus Proteins; Xenopus laevis
PubMed: 34784177
DOI: 10.1021/acs.biochem.1c00465 -
Poultry Science Jan 1979A morphological and histochemical study has been made of the primordial and early growing oocytes in the ovaries of crow (Corvus splendens) and common myna (Acridotheres... (Comparative Study)
Comparative Study
A morphological and histochemical study has been made of the primordial and early growing oocytes in the ovaries of crow (Corvus splendens) and common myna (Acridotheres tristis). The primordial oocytes in the myna ovary are loosely arranged in groups or nests, whereas in crow they form compact nests surrounded by highly vascularized connective tissue bands or lie in layers beneath the surface epithelium. The primordial oocytes in both the species are surrounded by flat granulosa cells whose number, shape, and cytochemical properties change with the initiation of growth. The oocyte nucleus shows a single basophilic nucleolus and thick diplotene chromosomes. With the initiation of growth, the number of nucleoli increases; simultaneously the chromosomes attain lampbrush configuration. Crescent-shaped Balbiani's vitelline body consists of ribonucleoproteins, lipoproteins, and phospholipids. The amount of these substances increases with the oocyte growth. The nature of proteins and lipids in the ooplasm and follicular epithelium also changes with the oocyte growth. Some randomly distributed protein bodies are also present in the ooplasm of primordial follicles. They disappear with the initiation of oocyte growth. The enzyme activities of acid phosphatase, NADP-diaphorase and NAD-diaphorase, also increase in the Balbiani's vitelline body with the oocyte growth. Alkaline phosphatase and delta 5-3 beta-HSDH activities are not seen. The possible functional significance of these morphological and histochemical changes has been discussed in relation to the initiation of growth in quiescent oocytes.
Topics: Acid Phosphatase; Animals; Birds; Dihydrolipoamide Dehydrogenase; Female; Histocytochemistry; Lipids; Oocytes; Ovum; Species Specificity
PubMed: 471889
DOI: 10.3382/ps.0580225 -
PloS One 2013Hermes is an RNA-binding protein that we have previously reported to be found in the ribonucleoprotein (RNP) particles of Xenopus germ plasm, where it is associated with...
Hermes is an RNA-binding protein that we have previously reported to be found in the ribonucleoprotein (RNP) particles of Xenopus germ plasm, where it is associated with various RNAs, including that encoding the germ line determinant Nanos1. To further define the composition of these RNPs, we performed a screen for Hermes-binding partners using the yeast two-hybrid system. We have identified and validated four proteins that interact with Hermes in germ plasm: two isoforms of Xvelo1 (a homologue of zebrafish Bucky ball) and Rbm24b and Rbm42b, both RNA-binding proteins containing the RRM motif. GFP-Xvelo fusion proteins and their endogenous counterparts, identified with antisera, were found to localize with Hermes in the germ plasm particles of large oocytes and eggs. Only the larger Xvelo isoform was naturally found in the Balbiani body of previtellogenic oocytes. Bimolecular fluorescence complementation (BiFC) experiments confirmed that Hermes and the Xvelo variants interact in germ plasm, as do Rbm24b and 42b. Depletion of the shorter Xvelo variant with antisense oligonucleotides caused a decrease in the size of germ plasm aggregates and loosening of associated mitochondria from these structures. This suggests that the short Xvelo variant, or less likely its RNA, has a role in organizing and maintaining the integrity of germ plasm in Xenopus oocytes. While GFP fusion proteins for Rbm24b and 42b did not localize into germ plasm as specifically as Hermes or Xvelo, BiFC analysis indicated that both interact with Hermes in germ plasm RNPs. They are very stable in the face of RNA depletion, but additive effects of combinations of antisense oligos suggest they may have a role in germ plasm structure and may influence the ability of Hermes protein to effectively enter RNP particles.
Topics: Animals; Carrier Proteins; Female; Germ Cells; Mutation; Oligodeoxyribonucleotides, Antisense; Oocytes; Protein Binding; Protein Interaction Domains and Motifs; Protein Interaction Mapping; Protein Transport; Ribonucleoproteins; Two-Hybrid System Techniques; Xenopus; Xenopus Proteins
PubMed: 24265795
DOI: 10.1371/journal.pone.0080077 -
Journal of Anatomy Jun 2021In order to understand the mechanism of the endocannabinoid (eCB) signal, which has so far been shown to work in oocyte genesis and maturation, it is critical to clarify...
In situ localization of diacylglycerol lipase α and β producing an endocannabinoid 2-arachidonoylglycerol and of cannabinoid receptor 1 in the primary oocytes of postnatal mice.
In order to understand the mechanism of the endocannabinoid (eCB) signal, which has so far been shown to work in oocyte genesis and maturation, it is critical to clarify detailed localization of the eCB synthesizing enzyme molecules as well as receptors for eCBs in oocytes in the ovary in situ. For this purpose, diacylglycerol lipase (DGL) α and β are involved in the synthesis of an eCB 2-arachidonoylglycerol (2-AG). DGLα/β and the cannabinoid receptor 1 (CB1) for 2-AG were shown to be localized to the primary oocytes of postnatal mice using immuno-light and electron microscopy. It was found that two types of localization existed: first, immunoreactivities for DGLα and β were weakly detected throughout the ooplasm in light microscopy for which the intracellular membranes of vesicles forming tiny scattered aggregates were responsible. Secondly, DGLβ-immunoreactivity was distinctly confined to the nuage of Balbiani bodies and small nuage-derivative structures; both amorphous materials and membranes of vesicles were responsible for their localization. On the other hand, the weak immunoreactivity for CB1 was localized in a pattern similar to the first one for DGLs, but not found in a pattern for the Balbiani nuage. Two routes of functional exertion of 2-AG synthesized by DGLs were suggested from the two types of localization: one was that the eCB synthesized at all the sites of DGLs is released from the oocytes and exerts paracrine or autocrine effects on adjacent intra-ovarian cells as well as the oocytes themselves. The other was that the eCB synthesized within the nuage was involved in the modulation of the posttranscriptional processing of oocytes. Owing to the failure in the detection of CB1 in the Balbiani nuage, however, the validity of the latter possibility remains to be elucidated.
Topics: Animals; Endocannabinoids; Lipoprotein Lipase; Mice; Oocytes; Receptor, Cannabinoid, CB1
PubMed: 33398893
DOI: 10.1111/joa.13392 -
The Journal of Cell Biology Jul 2020Although mitochondrial DNA (mtDNA) is prone to accumulate mutations and lacks conventional DNA repair mechanisms, deleterious mutations are exceedingly rare. How the...
Although mitochondrial DNA (mtDNA) is prone to accumulate mutations and lacks conventional DNA repair mechanisms, deleterious mutations are exceedingly rare. How the transmission of detrimental mtDNA mutations is restricted through the maternal lineage is debated. Here, we demonstrate that mitochondrial fission, together with the lack of mtDNA replication, segregate mtDNA into individual organelles in the Drosophila early germarium. After mtDNA segregation, mtDNA transcription begins, which activates respiration. Mitochondria harboring wild-type genomes have functional electron transport chains and propagate more vigorously than mitochondria containing deleterious mutations in hetreoplasmic cells. Therefore, mtDNA expression acts as a stress test for the integrity of mitochondrial genomes and sets the stage for replication competition. Our observations support selective inheritance at the organelle level through a series of developmentally orchestrated mitochondrial processes. We also show that the Balbiani body has a minor role in mtDNA selective inheritance by supplying healthy mitochondria to the pole plasm. These two mechanisms may act synergistically to secure the transmission of functional mtDNA through Drosophila oogenesis.
Topics: Animals; Cell Respiration; DNA Replication; DNA, Mitochondrial; Drosophila melanogaster; Electron Transport; Electron Transport Chain Complex Proteins; Female; Gene Expression Regulation, Developmental; Genes, Mitochondrial; Genome, Mitochondrial; Male; Mitochondria; Mitochondrial Dynamics; Mutation; Oocytes; Oogenesis
PubMed: 32375181
DOI: 10.1083/jcb.201905160