Dataset Information

Egg proteome changes during fertilization in sterlet Acipenser ruthenus

ABSTRACT: Sturgeons, producers of black caviar, are one of the most valuable wildlife commodities on earth (Pikitch et al., 2005). They appeared approximately 200 million years ago, and are known as living fossils (Bemis et al., 1997). Sturgeon populations dramatically declined as a result of overfishing, poaching and habitat destruction (Birstein, 1993; Billard and Lecointre, 2001; Vidotto et al., 2013). International Union for Conservation of Nature and Natural Resources (IUCN) classified over 85% of sturgeon species as at risk of extinction, more than any other group of species. Therefore, artificial reproduction techniques have been developed for sturgeons to meet the demands of aquaculture and restocking programs. Basic knowledge regarding biology of reproduction such as egg activation and fertilization can help improving the efficiency of artificial reproduction. Fertilization happens when gametes, spermatozoon and egg, are fused together. Egg activation refers to a series of morphological and molecular changes that occur in the egg during and after fertilization. Egg activation is believed to prevent polyspermy, and may also establish a micro-environment to support embryo development (Wong and Wessel, 2006; Niksirat et al., 2015a). Although, earlier researchers have illustrated egg structure and its morphological changes during egg fertilization and activation in sturgeons (Cherr and Clark, 1982, 1985a; Linhart and Kudo, 1997; Debus et al., 2008; Zelazowska, 2010), there is still a lack of molecular knowledge to improve our understanding of this stage of reproduction in these animals. Eggs of sturgeon are released to aqueous environment, where they are fertilized, activated and develop an adhesive layer after contact with freshwater (Cherr and Clark, 1984; Vorobeva and Markov, 1999). Although, egg stickiness is necessary for eggs in nature to help them find a stable position by attaching to substrates such as gravel and stone and increase chance of survival until hatching (Riehl and Patzner, 1998), but can cause high rate of mortality during artificial incubation as a result of attachment of eggs together and subsequent suffocation in limited space of incubators. Using clay suspension in water as an activation medium has been proven to be effective to block egg stickiness in sturgeon eggs (Dettlaff et al., 1993). In recent years, label-free protein quantification techniques have been developed based on the presumption of linear proportionality between peptide mass peak signal intensity and concentration of a given peptide. This approach avoids the limitations and multiple preparation steps of methods such as those based on two-dimensional gel electrophoresis (Niksirat et al., 2015b). The aim of present study was to identify and quantify proteome changes of sterlet eggs during activation in different activation media, water and clay suspension, using label-free protein quantification technique.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Danio Rerio (zebrafish) (brachydanio Rerio)

TISSUE(S): Egg

SUBMITTER: Hamid Niksirat

LAB HEAD: Peter James

PROVIDER: PXD006232 | Pride | 2017-06-12

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	140528_Hamid_10S_C18_dil5_01.RAW	Raw
	140528_Hamid_11S_C18_dil5_01.RAW	Raw
	140528_Hamid_12S_C18_dil5_01.RAW	Raw
	140528_Hamid_13S_C18_dil5_01.RAW	Raw
	140528_Hamid_14S_C18_dil5_01.RAW	Raw

Items per page:

1 - 5 of 16

Publications

Quantification of egg proteome changes during fertilization in sterlet Acipenser ruthenus.

Niksirat Hamid H Andersson Liselotte L Golpour Amin A Chupani Latifeh L James Peter P

Biochemical and biophysical research communications 20170608 2

Eggs of sterlet are discharged outside into ambient aquatic environment where egg activation and fertilization occur. Effects of different activation media including freshwater and clay suspension on protein abundances of egg were quantified in sterlet Acipenser ruthenus. In-gel digestion and high resolution mass spectrometry were used for label-free protein quantification in the eggs of five females. No significant (p > 0.05) difference was found between protein abundances in eggs activated wit ...[more]

PMID: 28602699

Similar Datasets

Project description:The life cycle of flowering plants alternates between a diploid sporophytic and a haploid gametophytic generation. After fertilization of each the egg and central cells by one male gamete, the development of both fertilization products occurs coordinated with the maternally derived seed coat and carpel tissues forming the fruit. The reproduction program is likely to involve the concerted activity of many genes. To identify genes with specific functions during reproduction, we have analyzed the expression profile of more than 22,000 genes present on the Arabidopsis ATH1 microarray during three stages of flower and fruit development. We found 1,886 genes regulated during reproductive development and 1,043 genes that were specifically expressed during reproduction. When compared to cells from an Arabidopsis suspension culture, S-phase genes were underrepresented and G2 and M-phase genes were strongly enriched in the set of specific genes, indicating that important functions during reproduction are exerted in the G2 and M phases of the cell cycle. Many potential signaling components, such as receptor-like protein kinases, phosphatases, and transcription factors, were present in both groups of genes. Members of the YABBY, MADS box, and Myb transcription factor families were significantly overrepresented in the group of specific genes, revealing an important role of these families during reproduction. Furthermore, we found a significant enrichment of predicted secreted proteins smaller than 15 kD that could function directly as signaling molecules or as precursors for peptide hormones. Our study provides a basis for targeted reverse-genetic approaches aimed to identify key genes of reproductive development in plants. During evolution, both animals and plants developed distinct pathways of sexual reproduction. The haploid life cycle starts with the formation of the meiotic products, for instance arrested primary oocytes and spermatocytes in mammals or micro- and megaspores in plants. The haploid phase ends when a sperm cell fuses with an egg cell to form the diploid zygote, which initiates embryo development. In animals, gametes are derived from a population of germ cells committed early during development to their reproductive fate and which usually rest until the onset of sexual maturity. By contrast, stem cells in shoot apical meristems of flowering plants divide continuously during postembryonic development. Only after formation of vegetative organs, the meristem produces specialized reproductive organs in which meiosis occurs, the stamens and carpel-borne ovules of the flower (for review, see Bowman and Eshed, 2000; Carles and Fletcher, 2003). As early as 1790, Goethe suggested that floral organs are derived from leaves and adapted to their specialized tasks (Goethe, 1790). Animal gametes differentiate directly from the products of meiosis, whereas in plants the meiotic products (spores) typically undergo two or three mitotic divisions to give rise to the male or female gametophytes, respectively (Drews et al., 1998; Grossniklaus and Schneitz, 1998). In several plant species, including Arabidopsis, the female gametophyte consists of seven cells: three antipodal cells, two synergid cells, one egg cell, and one central cell that contains two polar nuclei (Drews et al., 1998; Grossniklaus and Schneitz, 1998). The mature male gametophyte comprises two sperm cells contained within a vegetative cell (McCormick, 1993; da Costa-Nunes and Grossniklaus, 2003). During fertilization one sperm cell nucleus fuses with the egg cell nucleus, giving rise to the diploid embryo, whereas the other sperm cell fuses with the homodiploid central cell nucleus, generating the triploid endosperm (Drews and Yadegari, 2002). Subsequent seed and fruit development are highly coordinated processes between the embryo and endosperm, as well as the maternally derived testa (seed coat) and carpels. Because of the significant economical importance of flowers, seeds, and fruits, the complex processes involved in plant reproduction are not only of academic interest but also of high economic relevance. Many genes have been identified genetically that have a function in plant reproduction. The establishment of floral organ identity and initiation of seed development have been studied extensively, and many regulators of these processes have been identified (McElver et al., 2001; Zik and Irish, 2003; for review, see Chaudhury et al., 2001). High-throughput RNA profiling technologies can now complement the genetic and molecular approaches to provide new insights into plant reproduction. In particular, oligonucleotide-based microarrays can produce reliable, high-quality data (Lockhart et al., 1996; Hennig et al., 2003) to establish new biological knowledge on the transcriptional programs that are active during developmental processes (Spellman et al., 1998; Harmer et al., 2000; Becker et al., 2003; Honys and Twell, 2003). Here, we report a comprehensive Affymetrix GeneChip analysis of the Arabidopsis transcriptome at key steps of reproductive development. We identified 1,043 genes that were specifically expressed during reproduction. Among those genes were many potential signaling components, such as receptor-like protein kinases, phosphatases, and transcription factors. In addition, we found a significant enrichment of predicted secreted proteins smaller than 15 kD that could function directly as signaling molecules or as precursors for peptide hormones. Our study provides the basis for targeted reverse-genetic approaches aimed to identify important regulators of reproductive development in plants. Experimenter name = Lars Hennig Experimenter phone = +41 1 632 22 44 Experimenter fax = +41 1 632 10 44 Experimenter department = Plant Science C Experimenter institute = Institute of Plant Sciences and Zurich-Basel Experimenter address = Swiss Federal Institute of Technology Experimenter address = ETH Center Experimenter address = Zurich Experimenter zip/postal_code = CH-8092 Experimenter country = Switzerland Keywords: organism_part_comparison_design;

Project description:The biology and physiology of Siberian sturgeon reproduction differ substantially from teleost fish. Seminal plasma support and protect the viability, motility and fertilizing capacity of spermatozoa by creating the optimal environment for the storage. The present study for the first time described in-depth proteomic characterization of Siberian sturgeon seminal plasma using gel based and gel-free proteomic approaches: LC-MS/MS, 2DE and 2D blue native (BN)/SDS-PAGE. LC-MS/MS allowed to identify 657 proteins, which were classified according to their functions and pathways. 2DE visualized 339 spots corresponding to 76 unique proteins (almost 60% were present in various proteoforms). For the first time we demonstrated interactions between seminal proteins; four (C1-C4) multiprotein complexes were identified after 2D BN/SDS-PAGE; composed of (C1) serotransferrin-2 (TF) and fish-egg lectin (FEL); (C2) serum albumin 2 (ALB) and retinol-binding protein 4 (RBP4); (C3) ALB and TF (C3) and (C4) apolipoprotein A-I (APOA1), riboflavin-binding protein (RfBP) and myoglobin (MB). BN-SDS-PAGE also revealed that immunoglobulin and TF formed homomultimeric (dimer, trimer, tetramer and pentamer) complexes in seminal plasma. ALB, TF, Beta-Ala-His dipeptidase, glyceraldehyde-3-phosphate dehydrogenase, IGH, APOA1, hemopexin, type-4 ice-structuring protein, FEL, RfBP and glycogen phosphorylase, liver form-like were selected as major seminal plasma proteins. The functional analysis of identified proteins indicated their involvement mainly in immune system process and response to stimulus, metabolism, vesicle-mediated transport, proteolysis, and catherin binding involved in cell-cell adhesion. Moreover, 67 proteins were associated with reproductive processes, including spermatogenesis, fertilization, acrosomal reaction and sperm motility. Comparative proteomic analysis of sturgeon seminal plasma showed common proteins with fish and human and 150 proteins specific for sturgeon which may reflect the specificity of sturgeon reproduction. To conclude, this integrative proteomic view lead to deeper insight into physiological function of seminal plasma and processes occurred in sturgeon reproductive tract.

Project description:The life cycle of flowering plants alternates between a diploid sporophytic and a haploid gametophytic generation. After fertilization of each the egg and central cells by one male gamete, the development of both fertilization products occurs coordinated with the maternally derived seed coat and carpel tissues forming the fruit. The reproduction program is likely to involve the concerted activity of many genes. To identify genes with specific functions during reproduction, we have analyzed the expression profile of more than 22,000 genes present on the Arabidopsis ATH1 microarray during three stages of flower and fruit development. We found 1,886 genes regulated during reproductive development and 1,043 genes that were specifically expressed during reproduction. When compared to cells from an Arabidopsis suspension culture, S-phase genes were underrepresented and G2 and M-phase genes were strongly enriched in the set of specific genes, indicating that important functions during reproduction are exerted in the G2 and M phases of the cell cycle. Many potential signaling components, such as receptor-like protein kinases, phosphatases, and transcription factors, were present in both groups of genes. Members of the YABBY, MADS box, and Myb transcription factor families were significantly overrepresented in the group of specific genes, revealing an important role of these families during reproduction. Furthermore, we found a significant enrichment of predicted secreted proteins smaller than 15 kD that could function directly as signaling molecules or as precursors for peptide hormones. Our study provides a basis for targeted reverse-genetic approaches aimed to identify key genes of reproductive development in plants. During evolution, both animals and plants developed distinct pathways of sexual reproduction. The haploid life cycle starts with the formation of the meiotic products, for instance arrested primary oocytes and spermatocytes in mammals or micro- and megaspores in plants. The haploid phase ends when a sperm cell fuses with an egg cell to form the diploid zygote, which initiates embryo development. In animals, gametes are derived from a population of germ cells committed early during development to their reproductive fate and which usually rest until the onset of sexual maturity. By contrast, stem cells in shoot apical meristems of flowering plants divide continuously during postembryonic development. Only after formation of vegetative organs, the meristem produces specialized reproductive organs in which meiosis occurs, the stamens and carpel-borne ovules of the flower (for review, see Bowman and Eshed, 2000; Carles and Fletcher, 2003). As early as 1790, Goethe suggested that floral organs are derived from leaves and adapted to their specialized tasks (Goethe, 1790). Animal gametes differentiate directly from the products of meiosis, whereas in plants the meiotic products (spores) typically undergo two or three mitotic divisions to give rise to the male or female gametophytes, respectively (Drews et al., 1998; Grossniklaus and Schneitz, 1998). In several plant species, including Arabidopsis, the female gametophyte consists of seven cells: three antipodal cells, two synergid cells, one egg cell, and one central cell that contains two polar nuclei (Drews et al., 1998; Grossniklaus and Schneitz, 1998). The mature male gametophyte comprises two sperm cells contained within a vegetative cell (McCormick, 1993; da Costa-Nunes and Grossniklaus, 2003). During fertilization one sperm cell nucleus fuses with the egg cell nucleus, giving rise to the diploid embryo, whereas the other sperm cell fuses with the homodiploid central cell nucleus, generating the triploid endosperm (Drews and Yadegari, 2002). Subsequent seed and fruit development are highly coordinated processes between the embryo and endosperm, as well as the maternally derived testa (seed coat) and carpels. Because of the significant economical importance of flowers, seeds, and fruits, the complex processes involved in plant reproduction are not only of academic interest but also of high economic relevance. Many genes have been identified genetically that have a function in plant reproduction. The establishment of floral organ identity and initiation of seed development have been studied extensively, and many regulators of these processes have been identified (McElver et al., 2001; Zik and Irish, 2003; for review, see Chaudhury et al., 2001). High-throughput RNA profiling technologies can now complement the genetic and molecular approaches to provide new insights into plant reproduction. In particular, oligonucleotide-based microarrays can produce reliable, high-quality data (Lockhart et al., 1996; Hennig et al., 2003) to establish new biological knowledge on the transcriptional programs that are active during developmental processes (Spellman et al., 1998; Harmer et al., 2000; Becker et al., 2003; Honys and Twell, 2003). Here, we report a comprehensive Affymetrix GeneChip analysis of the Arabidopsis transcriptome at key steps of reproductive development. We identified 1,043 genes that were specifically expressed during reproduction. Among those genes were many potential signaling components, such as receptor-like protein kinases, phosphatases, and transcription factors. In addition, we found a significant enrichment of predicted secreted proteins smaller than 15 kD that could function directly as signaling molecules or as precursors for peptide hormones. Our study provides the basis for targeted reverse-genetic approaches aimed to identify important regulators of reproductive development in plants. Experimenter name = Lars Hennig; Experimenter phone = +41 1 632 22 44; Experimenter fax = +41 1 632 10 44; Experimenter department = Plant Science C; Experimenter institute = Institute of Plant Sciences and Zurich-Basel; Experimenter address = Swiss Federal Institute of Technology; Experimenter address = ETH Center; Experimenter address = Zurich; Experimenter zip/postal_code = CH-8092; Experimenter country = Switzerland Experiment Overall Design: 6 samples were used in this experiment