Felipe Perecin

Universidade de São Paulo

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Flávio Vieira Meirelles

Universidade de São Paulo

105

Juliano Coelho da Silveira

Universidade de São Paulo

Juliano Rodrigues Sangalli

Universidade de São Paulo

Maite del Collado

Universidade de São Paulo

Gabriella Mamede Andrade

Universidade de São Paulo

Fabiana Fernandes Bressan

Universidade de São Paulo

Lawrence C. Smith

Université de Montréal

Christina R. Ferreira

Purdue University West Lafayette

Tiago Henrique Camara De

Universidade de São Paulo

Marcos Roberto Chiaratti

Universidade Federal de São Carlos

Cooperative Institutions

Universidade de São Paulo

248

Division of Undergraduate Education

247

Institute of Developmental Physiology

107

Universidade Brasil

Universidade Estadual Paulista (Unesp)

Universidade Estadual de Campinas (UNICAMP)

Hospital Universitário da Universidade de São Paulo

Université de Montréal

Fundação de Estudo e Pesquisa em Medicina Veterinária e Zootecnia

Universidade Federal de São Paulo

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Steps forward canine primordial germ cell isolation and characterization

Reproduction Abstracts (2014)

Souza Aline Fernanda de Naira Caroline Godoy Pieri Juliana Barbosa Cassals Ana Carolina Furlanetto Mançanares Kelly Cristine Santos Roballo

Searchable abstracts of presentations at key conferences on reproductive biology and medicine ISSN 2052-1472 (online)

Isolation

Characterization

10.1530/repabs.1.p195

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Association of in vivo fertility of bulls with in vivo production of embryos

Animal Reproduction (2018)

Guilherme Machado Zanatta Flávia Morag Elliff A Brid Gabriel Armond Crepaldi Rodolfo Daniel Mingoti

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Exogenous OCT4 and SOX2 Contribution to In Vitro Reprogramming in Cattle

Biomedicines (2023)

Lucas Simões Machado Camila Martins Borges Marina Amaro de Lima Juliano Rodrigues Sangalli Jacinthe Therrien

Mechanisms of cell reprogramming by pluripotency-related transcription factors or nuclear transfer seem to be mediated by similar pathways, and the study of the contribution of OCT4 and SOX2 in both processes may help elucidate the mechanisms responsible for pluripotency. Bovine fibroblasts expressing exogenous OCT4 or SOX2, or both, were analyzed regarding the expression of pluripotency factors and imprinted genes H19 and IGF2R, and used for in vitro reprogramming. The expression of the H19 gene was increased in the control sorted group, and putative iPSC-like cells were obtained when cells were not submitted to cell sorting. When sorted cells expressing OCT4, SOX2, or none (control) were used as donor cells for somatic cell nuclear transfer, fusion rates were 60.0% vs. 64.95% and 70.53% vs. 67.24% for SOX2 vs. control and OCT4 vs. control groups, respectively; cleavage rates were 66.66% vs. 81.68% and 86.47% vs. 85.18%, respectively; blastocyst rates were 33.05% vs. 44.15% and 52.06% vs. 44.78%, respectively. These results show that the production of embryos by NT resulted in similar rates of in vitro developmental competence compared to control cells regardless of different profiles of pluripotency-related gene expression presented by donor cells; however, induced reprogramming was compromised after cell sorting.

Reprogramming

Homeobox protein NANOG

XIST

Rex1

10.3390/biomedicines11092577

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Citations (2)

Low levels of exosomal-miRNAs in maternal blood are associated with early pregnancy loss in cloned cattle

Scientific Reports (2017)

Tiago Henrique Camara De Juliano Coelho da Silveira Rafael Vilar Sampaio Juliano Rodrigues Sangalli M. L. F. Oliveira

Abstract Nuclear reprogramming mediated by somatic cell nuclear transfer (SCNT) has many applications in medicine. However, animal clones show increased rates of abortion and reduced neonatal viability. Herein, we used exosomal-miRNA profiles as a non-invasive biomarker to identify pathological pregnancies. MiRNAs play important roles in cellular proliferation and differentiation during early mammalian development. Thus, the aim of this study was to identify exosomal-miRNAs in maternal blood at 21 days of gestation that could be used for diagnosis and prognosis during early clone pregnancies in cattle. Out of 40 bovine-specific miRNAs, 27 (67.5%) were with low abundance in the C-EPL (Clone - Early pregnancy loss) group compared with the C-LTP (Clone - Late pregnancy) and AI-LTP (Artificial Insemination - Late pregnancy) groups, which had similar miRNAs levels. Bioinformatics analysis of the predicted target genes demonstrated signaling pathways and functional annotation clusters associated with critical biological processes including cell proliferation, differentiation, apoptosis, angiogenesis and embryonic development. In conclusion, our results demonstrate decreased exosomal-miRNAs in maternal blood at 21 days of gestation in cloned cattle pregnancies that failed to reach term. Furthermore, the predicted target genes regulated by these 27 miRNAs are strongly associated with pregnancy establishment and in utero embryonic development.

Reprogramming

Early Pregnancy Loss

10.1038/s41598-017-14616-1

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Citations (41)

The embryo non-invasive pre-implantation diagnosis era: how far are we?

Animal Reproduction (2023)

Maite del Collado Gabriella Mamede Andrade N Gonçalves Samuel Fortini Felipe Perecin

Advancements in assisted reproduction (AR) methodologies have allowed significant improvements in live birth rates of women who otherwise would not be able to conceive. One of the tools that allowed this improvement is the possibility of embryo selection based on genetic status, performed via preimplantation genetic testing (PGT). Even though the widespread use of PGT from TE biopsy helped to decrease the interval from the beginning of the AR intervention to pregnancy, especially in older patients, in AR, there are still many concerns about the application of this invasive methodology in all cycles. Therefore, recently, researchers started to study the use of cell free DNA (cfDNA) released by the blastocyst in its culture medium to perform PGT, in a method called non-invasive PGT (niPGT). The development of a niPGT would bring the diagnostics power of conventional PGT, but with the advantage of being potentially less harmful to the embryo. Its implementation in clinical practice, however, is under heavy discussion since there are many unknowns about the technique, such as the origin of the cfDNA or if this genetic material is a true representative of the actual ploidy status of the embryo. Available data indicates that there is high correspondence between results observed in TE biopsies and the ones observed from cfDNA, but these results are still contradictory and highly debatable. In the present review, the advantages and disadvantages of niPGT are presented and discussed in relation to tradition TE biopsy-based PGT. Furthermore, there are also presented some other possible non-invasive tools that could be applied in the selection of the best embryo, such as quantification of other molecules as quality biomarkers, or the use artificial intelligence (AI) to identify the best embryos based on morphological and/or morphokitetic parameters.

10.1590/1984-3143-ar2023-0069

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Mice born to females with oocyte-specific deletion of mitofusin 2 have increased weight gain and impaired glucose homeostasis

Molecular Human Reproduction (2020)

Bruna Martins Garcia Thiago Simões Machado Karen Freire Carvalho Patrícia Nolasco Ricardo Perecin Nociti

Abstract Offspring born to obese and diabetic mothers are prone to metabolic diseases, a phenotype that has been linked to mitochondrial dysfunction and endoplasmic reticulum (ER) stress in oocytes. In addition, metabolic diseases impact the architecture and function of mitochondria-ER contact sites (MERCs), changes which associate with mitofusin 2 (MFN2) repression in muscle, liver and hypothalamic neurons. MFN2 is a potent modulator of mitochondrial metabolism and insulin signaling, with a key role in mitochondrial dynamics and tethering with the ER. Here, we investigated whether offspring born to mice with MFN2-deficient oocytes are prone to obesity and diabetes. Deletion of Mfn2 in oocytes resulted in a profound transcriptomic change, with evidence of impaired mitochondrial and ER function. Moreover, offspring born to females with oocyte-specific deletion of Mfn2 presented increased weight gain and glucose intolerance. This abnormal phenotype was linked to decreased insulinemia and defective insulin signaling, but not mitochondrial and ER defects in offspring liver and skeletal muscle. In conclusion, this study suggests a link between disrupted mitochondrial/ER function in oocytes and increased risk of metabolic diseases in the progeny. Future studies should determine whether MERC architecture and function are altered in oocytes from obese females, which might contribute toward transgenerational transmission of metabolic diseases.

MFN2

10.1093/molehr/gaaa071

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Mass spectrometry fingerprinting of media used for in vitro production of bovine embryos

Rapid Communications in Mass Spectrometry (2009)

Christina R. Ferreira Gustavo H.M.F. Souza Maria Francesca Riccio Rodrigo Ramos Catharino J. H. F. Pontes

Abstract Using the bovine species as a biological model, direct infusion chip‐based nano‐electrospray ionization mass spectrometry (nano‐ESI‐MS) fingerprinting in the positive ion mode is used to obtain fast chemical profiles of media used for in vitro production of bovine embryos. Nano‐ESI‐MS fingerprinting is useful for characterization and routine quality control requiring no sample pre‐separation, being able to differentiate four different media (IVM, IVF, SOF and HSOF) via principal component analysis (PCA). For media stored at +4°C for up to 45 days, no significant ( p > 0.05) variation was observed in cleavage and blastocyst rate development, as well as in the nano‐ESI‐MS chemical profiles. For media exposed to a heat shock (60°C for 3 h), no significant decrease ( p > 0.05) in embryo development rates was observed, but nano‐ESI‐MS profiles were quite distant from fresh control media in the PCA. For frozen media (−70°C for 2 months), again no significant variation ( p > 0.05) in embryo development was noticed, but nano‐ESI‐MS profiles from all media were significantly affected. These results indicate that nano‐ESI(+)‐MS fingerprinting was able to characterize different media based on their specific chemical profile. The technique seems therefore applicable as a routine quality control assay, detecting, for example, compositional changes after temperature variations that may affect post‐transfer embryo viability. Copyright © 2009 John Wiley & Sons, Ltd.

Control sample

10.1002/rcm.3995

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Citations (21)

Bovine in vitro oocyte maturation and embryo culture in liquid marbles 3D culture system

PLoS ONE (2023)

Giuliana de Avila Ferronato Carolina Mônica dos Santos Paola Maria da Silva Rosa Alessandra Bridi Felipe Perecin

Despite the advances in in vitro embryo production (IVP) over the years, the technique still has limitations that need to be overcome. In cell cultures, it is already well established that three-dimensional culture techniques are more physiological and similar to the in vivo development. Liquid marble (LM) is a three-dimensional system based on the use of a hydrophobic substance to create in vitro microbioreactors. Thus, we hypothesized that the LM system improves bovine in vitro oocyte maturation and embryo culture. In experiment I, bovine cumulus-oocyte complexes (COCs) were placed for in vitro maturation for 22h in two different groups: control (conventional 2D culture) and LM (three-dimensional culture). We found that oocyte nuclear maturation was not altered by the LM system, however it was observed a decrease in expression of genes important in the oocyte maturation process in cumulus cells of LM group ( BCL2 , EIF4E , and GAPDH ). In experiment II, the COCs were conventionally matured and fertilized, and for culture, they were divided into LM or control groups. There was a decrease in blastocyst rate and cell counting, a down-regulation of miR-615 expression, and an increase in the DNA global methylation and hydroxymethylation in embryos of LM group. Therefore, for the bovine in vitro embryo production, this specific three-dimensional system did not present the advantages that we expected, but demonstrated that the embryos changed their development and epigenetics according to the culture system.

In vitro maturation

10.1371/journal.pone.0284809

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Efeitos da obesidade sobre a expressão de genes relacionados ao metabolismo lipídico e de proteínas carreadoras de ácidos graxos em ovários murinos

Jaqueline Natalia Pizzaia Felipe Perecin Maite Barrondo Del Collado

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Corpus luteum presence in the bovine ovary increase intrafollicular progesterone concentration: consequences in follicular cells gene expression and follicular fluid small extracellular vesicles miRNA contents

Journal of Ovarian Research (2024)

Paola Maria da Silva Rosa Alessandra Bridi Giuliana de Ávila Ferronato Cibele M. Prado Natália Marins Bastos

Abstract Background It is well described that circulating progesterone (P4) plays a key role in several reproductive events such as oocyte maturation. However, during diestrus, when circulating P4 is at the highest concentrations, little is known about its local impact on the follicular cells such as intrafollicular P4 concentration due to corpus luteum (CL) presence within the same ovary. Based on that, our hypothesis is that the CL presence in the ovary during diestrus alters intrafollicular P4 concentrations, oocyte competence acquisition, follicular cells gene expression, and small extracellular vesicles (sEVs) miRNAs contents. Results P4 hormonal analysis revealed that ipsilateral to the CL follicular fluid (iFF) presented higher P4 concentration compared to contralateral follicular fluid (cFF). Furthermore, oocyte maturation and miRNA biogenesis pathways transcripts ( ADAMTS-1 and AGO2 , respectively) were increased in cumulus and granulosa cells of iFF, respectively. Nevertheless, a RT-PCR screening of 382 miRNAs showed that three miRNAs were upregulated and two exclusively expressed in sEVs from iFF and are predicted to regulate cell communication pathways. Similarly, seven miRNAs were higher and two exclusively expressed from cFF sEVs and are predicted to modulate proliferation signaling pathways. Conclusion In conclusion, intrafollicular P4 concentration is influenced by the presence of the CL and modulates biological processes related to follicular cell development and oocyte competence, which may influence the oocyte quality. Altogether, these results are crucial to improve our knowledge about the follicular microenvironment involved in oocyte competence acquisition.

Follicular fluid

Granulosa cell

10.1186/s13048-024-01387-3

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Citations (5)