A pig multi-tissue normalised cDNA library: large-scale sequencing, cluster analysis and 9K micro-array resource generation

Bonnet, Agnès; Iannuccelli, Eddie; Hugot, Karine; Benne, Francis; Bonaldo, Maria F.; Soares, Marcelo B.; Hatey, François; Tosser‐Klopp, Gwenola

doi:10.1186/1471-2164-9-17

Cited by 23 publications

(30 citation statements)

References 41 publications

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“…The existence of wide spectra of genes whose expres- sion is typical only of oocytes was demonstrated by Bonnet et al (2008). nNOS mRNA was detected in oocytes, but not in their granulosa or cumulus cells.…”

Section: Discussionmentioning

confidence: 99%

Expression and localization of nitric oxide synthase isoforms during porcine oocyte growth and acquisition of meiotic competence

Eva¹,

Sedmı́ková²,

Petr³

et al. 2009

CZECH J ANIM SCI

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The intensive development of biotechnologies such as in vitro embryo production and cloning by nuclear transfer requires a great number of oocytes with fully-developed meiotic competence.During in vitro culture these oocytes are able to reach the stage of second meiotic metaphase and complete their meiotic maturation. However, the number of these oocytes in the ovary is limited. ABSTRACT: Reproduction biotechnologies depend on the use of fully meiotically competent oocytes. Growing oocytes without full meiotic competence are an interesting potential source due to their quantity, but the mechanisms regulating the processes of acquisition of meiotic competence have not been clarified to date. Nitric oxide synthase (NOS) and its product, nitric oxide (NO), may possibly play a role. Understanding the precise NO regulatory mechanism is therefore important for the development of in vitro growth methods. The objective of this work was to detect changes in the expression of NOS isoforms and their mRNA expression and changes in the intracellular localization of separate NOS isoforms during the growth period of the porcine oocyte, and also to determine whether these changes are related to the process of meiotic competence acquisition. mRNA for all NOS isoforms was already detected in oocytes at the beginning of their growth and was present in them until they completed their growth period. mRNA for iNOS and eNOS was also observed in granulosa and cumulus cells from these oocytes. But nNOS mRNA was not demonstrated in these types of cells. Pig oocytes and their surrounding cells contained all NOS proteins. Their amounts increased and localization changed with the acquisition of meiotic competence. nNOS was localized mainly in the cortex in meiotically incompetent oocytes, while meiotically competent oocytes contained nNOS in the nucleus as well. iNOS protein was distributed in the cytoplasm and nucleus in all oocytes, and meiotically incompetent oocytes contained iNOS in the nucleolus as well. eNOS protein was distributed in oocytes in the form of fine granules with a strong fluorescence signal. Protein was concentrated in the nuclear area in meiotically incompetent oocytes and also in the periphery in oocytes with partially and fully-developed meiotic competence. All these findings indicate that NOS isoforms may significantly influence the acquisition of meiotic competence in porcine oocytes.

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Section: Discussionmentioning

confidence: 99%

Expression and localization of nitric oxide synthase isoforms during porcine oocyte growth and acquisition of meiotic competence

Eva¹,

Sedmı́ková²,

Petr³

et al. 2009

CZECH J ANIM SCI

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“…The micro-arrays contained PCR products from 2849 pig cDNA clones coming from 1697 clones selected after SSH/macroarray experiments, 1056 clones of the AGENAE pig normalized multi-tissue cDNA library (Bonnet et al 2008) and 96 clones used as controls. The multi-tissue library was used to allow a proper normalization of the data.…”

Section: Micro-arrays Design and Hybridizationmentioning

confidence: 99%

In vivo gene expression in granulosa cells during pig terminal follicular development

Bonnet¹,

Cao²,

SanCristobal³

et al. 2008

Reproduction

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Ovarian antral follicular development is clearly dependent on pituitary gonadotrophins FSH and LH. Although the endocrine mechanism that controls ovarian folliculogenesis leading to ovulation is quite well understood, the detailed mechanisms and molecular determinants in the different follicular compartments remain to be clarified. The aim of this study was to identify the genes differentially expressed in pig granulosa cells along the terminal ovarian follicle growth, to gain a comprehensive view of these molecular mechanisms. First, we developed a specific micro-array using cDNAs from suppression subtractive hybridization libraries (345 contigs) obtained by comparison of three follicle size classes: small, medium and large antral healthy follicles. In a second step, a transcriptomic analysis using cDNA probes from these three follicle classes identified 79 differentially expressed transcripts along the terminal follicular growth and 26 predictive genes of size classes. The differential expression of 18 genes has been controlled using real-time PCR experiments validating the micro-array analysis. Finally, the integration of the data using Ingenuity Pathways Analysis identified five gene networks providing descriptive elements of the terminal follicular development. Specifically, we observed: (1) the down-expression of ribosomal protein genes, (2) the genes involved in lipid metabolism and (3) the down-expression of cell morphology and ion-binding genes. In conclusion, this study gives new insight into the gene expression during pig terminal follicular growth in vivo and suggested, in particular, a morphological change in pig granulosa cells accompanying terminal follicular growth. Reproduction (2008) 136 211-224

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“…The genotyping was realized with 170 microsatellite markers and LM expression profiles were obtained with a microarray of 9000 PCR products from a multi-tissues library (Bonnet et al, 2008). 272 genes were identified to be genetically regulated by 335 eQTL (Liaubet et al, 2011).…”

Section: Eqtl In Pigmentioning

confidence: 99%

Recent advances in omic technologies for meat quality management

et al. 2015

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A pig multi-tissue normalised cDNA library: large-scale sequencing, cluster analysis and 9K micro-array resource generation

Cited by 23 publications

References 41 publications

Expression and localization of nitric oxide synthase isoforms during porcine oocyte growth and acquisition of meiotic competence

Expression and localization of nitric oxide synthase isoforms during porcine oocyte growth and acquisition of meiotic competence

In vivo gene expression in granulosa cells during pig terminal follicular development

Recent advances in omic technologies for meat quality management

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