Project #1: Epigenetic-miRNA circuitry directing neural crest EMT/MET: Neural crest is a population of multipotent stem cells that originate in the dorsal part of the forming neural tube. Around the time of neural tube closure, they lose their epithelial characteristics and delaminate, by process known as epithelial-to-mesenchymal transition (EMT), acquiring migratory ability. Once neural crest cells reach their destination, they differentiate into numerous derivatives including neurons and glia of sensory ganglia. In a manner that is essentially the reverse of EMT, the formation of sensory ganglia requires the coalescence of neural crest cells via a process of mesenchymal-to-epithelial transition (MET). Both embryonic EMT and MET bear similarities to the molecular pathways and cellular changes undertaken by cancer cells during metastasis and establishment of secondary tumors. In recent years, there has been demonstrated that epigenetic-miRNA circuitries regulate EMT in various type of cancerous cells. However, the in vivo study of MET in cancer cells is very complex and unpredictable. In contrast, neural crest development is highly regulated and predictable, since the migratory pathways and process of condensation into sensory ganglia are well characterized. Based on this, we are studding the existence of a reversible epigenetic-microRNA regulatory networks that may occur during transitional states of neural crest cells delamination (EMT) and sensory ganglia coalescence (MET).

During embryo development, exit from pluripotency and sequential activation of distinct differentiation programs must tightly controlled to coordinate cell fate decisions with morphogenetic processes. Around the time of gastrulation ectodermal cells begin to segregate the neural and neural plate border (neural crest and placode progenitors) by activation/repressing a sequential set of genes. The miRNAs are a specific subgroup of ncRNA that are involved in myriad cellular events including the balance between proliferation and differentiation as well as the spatial-temporal modulation of gene expression during development. Development is often assumed to be hardwired in the genome, but several lines of evidence indicate that it is susceptible to environmental modulation with potential long-term consequences. In this context, it is well known that vitamins supplementation is required during the periconceptional periods in human. Specifically, folate and vitamin C (ascorbic acid) acts as the yin and yang on DNA methylation, been the first required to generate the donor of methyl groups (S-adenosylmethionine or SAM), and the second as a cofactor for demethylases (TETs). Approximately 50% of miRNA genes are embedded or associated with CpGs islands, and their expression is most often regulated by methylation of cytosines therein. Based on this, we are studding the role of vitamin C and folate availability on the epigenetic regulation of key microRNAs involved on the territorial restriction of neural and neural plate progenitors.

Folate is an essential compound for many body functions. Humans are unable to synthesize folate; thus, it has to be incorporated in the diet to meet their metabolic needs. Thus, folate deficiency in the early stages of development leads to severe neural tube and neural crest defects such as spina bifida and craniofacial disorders. Alcohol is well known to be teratogenic, causing developmental abnormalities. Fetal alcohol syndrome is characterized as pre- and postnatal growth retardation, central nerve system abnormalities, distinct facial features, and intellectual retardation. Although both folate deficiency and alcohol exposition developmental toxicity has been primarily attributed to maternal alcohol exposure during pregnancy, recent studies in human and animal models have suggested that fathers may contribute to developmental abnormalities. Very recent studies demonstrated that certain paternal traits can be inherited by the offspring, suggesting that epigenetic inheritance can occur through the sperm. The search for this “Lamarckian factors” in the sperm that transmit acquired phenotypes to the offspring has recently focused on RNAs and their modifications, particularly on small non-coding RNAs (sncRNAs) including microRNAs (miRNAs) and tRNA-derived small RNAs (tsRNAs). These sncRNAs can ultimate regulate DNA methylation, histone modifications, and mRNA transcription and are also emerging as a modulator of offspring development. Based on this in our lab we are studying the paternal effect of alcohol exposition and folate deficiency on the dysregulations of sncRNA modifications and content in the sperm that may produce intergenerational developmental defects.

The regulation of the proliferation of germ cells is a fundamental process for the differentiation of the gonads as well as the reproduction of any species. This regulation is carried out by the optimal balance between proliferation, necessary for the production of an adequate number of gametes, and apoptosis or programmed cell death, necessary for the elimination of defective gametes and the control of proliferation. The loss of this balance can produce the hyper-proliferation of germ cells, producing testicular or ovarian neoplasms or cancer, or on the other hand, an excess in apoptosis would lead to the sterility of the individual. This balance is regulated at the endocrine and molecular level. This is why we are interested in knowing the molecular, epigenetic and endocrine mechanisms that regulate this fundamental process for reproductive success.

Recent studies reveal the existence of a possible cross-talk mechanism between HPT and HPG, where, for example, thyroid hormones (HTs) would positively regulate androgen synthesis at environmentally relevant concentrations in the masculinization observed in fish and frogs exposed to these hormones. We intend to determine the effects of HTs in the gonadal development of two models of aquatic vertebrates: Oryzias latipes and Silurana tropicalis, to elucidate the molecular mechanisms involved in this cross-regulation. With this project, the regulation of HTs in the synthesis of androgens in a key development process, such as gonadal differentiation, will be known, establishing an interesting molecular and endocrinological comparative proposal.

Bernardi YE, Sanchez-Vasquez E, Márquez RB, Piacentino ML, Urrutia H, Rossi I, Alcântara Saraiva KL, Pereira-Neves A, Ramirez MI, Bronner ME, de Miguel N, Strobl- Mazzulla PH. miR-203 secreted in extracellular vesicles mediates the communication between neural crest and placode cells required for trigeminal ganglia formation. Plos Biology. En prensa. https://doi.org/10.1371/journal.pbio.3002074

Boan AF, Delgadin TH, Canosa LF, Fernandino JI. Loss of function in somatostatin receptor 5 has no impact on the growth of medaka fish due to compensation by the other paralogs. General and Comparative Endocrinology 351, 114478. 2024. https://doi.org/10.1016/j.ygcen.2024.114478

Lopez-Landavery EA, Urquizo-Rosado Á, Saavedra-Flores A, Tapia-Morales S, Fernandino JI, Zelada-Mázmela E. Cellular and transcriptomic response to pathogenic and non-pathogenic Vibrio parahaemolyticus strains causing acute hepatopancreatic necrosis disease (AHPND) in Litopenaeus vannamei. Fish and Shellfish Immunology 148, 109472. 2024. https://doi.org/10.1016/j.fsi.2024.109472

Sacatturice LA, Vázquez N, Strobl-Mazzulla PH. miR-137 conferred robustness to the territorial restriction of the neural plate border. Development. En prensa. https://doi.org/10.1242/dev.202344

Torres-Martínez A, Hattori RS, Fernandino JI, Somoza MG, Hung SD, Yamamoto Y, Strüssmann CA. Temperature- and genotype-dependent stress response and activation of the hypothalamus-pituitary-interrenal axis during temperature-induced sex reversal in pejerrey Odontesthes bonariensis, a species with genotypic and environmental sex determination. Molecular and Celular Endocrinology 582, 112114. 2024. doi:10.1016/j.mce.2023.112114

Alata Jimenez NA, Casterllano M, Santillan EM, Boulias K, Boan A, Padilla LF, Fernandino JI, Greer EL, Tosar JP, Cochella L, Strobl-Mazzulla PH. Paternal methotrexate exposure affects sperm small RNA content and causes craniofacial defects in the offspring. Nature Communications  14, 1617. 2023. https://doi.org/10.1038/s41467-023-37427-7

Carriquiriborde P, Fernandino JI, López CG, de San Benito E, Gutierrez-Villagomez JM, Cristos D, Trudeay VL, Somoza GM. Atrazine alters early sexual development of the South American silverside, Odontesthes bonariensis. Aquatic Toxicology 254, 106366. 2023. doi: doi:10.1016/j.aquatox.2022.106366

Castañeda-Cortés DC, Rosa IF, Boan AF, Marrone D, Pagliaro N, Oliveira MA, Rodrigues MS, Doretto LB, Silva C, Tavares-Júnior J, Costa DF, Dodds MS, Strobl-Mazzulla PH, Langlois VS, Nóbrega RH, Fernandino JI. Thyroid axis participates in high-temperature-induced male sex reversal through its activation by the stress response. Cellular and Molecular Life Sciences 80, 253. 2023. https://doi.org/10.1007/s00018-023-04913-6

Marín A, Alonso AM, Delgadín TH, López-Landavery EA, Lise J.A.R.A. Cometivos, Saavedra-Flores A, Reyes-Flores LE, Yzásiga-Barrera C, Fernandino JI, Zelada-Mázmela E. Analysis of truncated growth hormone receptor 1 in the differential growth of fine flounder Paralichthys adspersus. Aquaculture 574. 739691. 2023. https://doi.org/10.1016/j.aquaculture.2023.739691

Moreno Acosta OD, Boan AF, Hattori RS, Fernandino JI. Notch pathway is required for protection against heat stress in spermatogonial stem cells in medaka. Fish Physiology and Biochemistry 49, 487-500. 2023. https://doi.org/10.1007/s10695-023-01200-w

Alata Jimenez N, Strobl-Mazzulla P.Folate carrier deficiency drives differential methylation and enhanced cellular potency in the neural plate border. Frontiers in Cell and Developmental Biology 10, 834625.2022. https://doi.org/10.3389/fcell.2022.834625

Delbes G, Blázquez M, Fernandino JI, Grigorova P, Hales BF, Metcalfe C, Navarro-Martín L, Parent L, Robaire B, Rwigemera A, Van Der Kraak G, Wade M, Marlatt V. Effects of endocrine disrupting chemicals on gonad development: Mechanistic insights from fish and mammals. Environmental Research 204B,  112040. 2022. https://doi.org/10.1016/j.envres.2021.112040

Goikoetxea A, Servill A, Houdelet C, Mouchel O, Hermet S, Clota F, Aerts J, Fernandino JI, Allal F, Vandeputte, Blondeau‑Bidet E, Geffroy B. Natural cortisol production is not linked to the sexual fate of European sea bass.  Fish Physiol Biochem 48, 1117-1135. 2022.  https://doi.org/10.1007/s10695-022-01104-1

López-Landavery EA, Corona-Herrera GA, Santos-Rojas LE, Herrera-Castilo NM, Delgadin TH, Tapia-Morales S, González-Martínez S, Reyes–Flores LE, Marín A, Yzásiga–Barrera CG, Fernandino JI, Zelada–Mázmela E. Non-invasive sex genotyping of paiche Arapaima gigas by qPCR: An applied bioinformatic approach for identifying sex differences. Aquaculture 546, 737388 2022. https://doi.org/10.1016/j.aquaculture.2021.737388

Arias Padilla LF, Castañeda-Cortés DC, Rosa IF, Moreno Acosta OD, Hattori RS, Nóbrega RH, Fernandino JI. Cystic proliferation of germline stem cells is necessary to reproductive success and normal mating behavior in medaka. Elife 10, e62757.2021. https://doi.org/10.7554/eLife.62757

Bernardi Y, Strobl-Mazzulla PH. What we can learn from embryos to understand the mesenchymal-to-epithelial transition in tumor progression. Biochemical Journal 478, 1809-1825. 2021. https://doi.org/10.1042/BCJ20210083

Butzge AJ, Yoshinaga TT, Moreno Acosta OD, Fernandino JI, Sanches EA, Tabata YA, de Oliveira C, Takahashi NS, Hattori RS. Early warming stress on rainbow trout juveniles impairs male reproduction but contrastingly elicits intergenerational thermotolerance. Scientifc Reports 11, 17053. 2021.  https://doi.org/10.1038/s41598-021-96514-1.

Castañeda-Cortés DC, Fernandino JI. Stress and sex determination in fish: From brain to gonads. The International Journal of Developmental Biology 65, 207-214. 2021. https://doi:10.1387/ijdb.200072jf 

Lizarraga A, Muñoz D, Strobl-Mazzulla PH, de Miguel N. Toward incorporating epigenetics into regulation of gene expression in the parasite Trichomonas vaginalis. Molecular Microbiology. En prensa. https://doi.org/10.1111/mmi.14704

Marín A, López–Landavery E, González–Martinez S, Reyes-Flores L, Corona-Herrera G, Tapia-Morales S, Yzásiga-Barrera C, Fernandino JI, Zelada-Mázmela E. The complete mitochondrial genome of the fine flounder Paralichthys adspersus revealed by next-generation sequencing.  Mitochondrial DNA Part B 6, 2785-2787. 2021. https://doi.org/10.1080/23802359.2021.1914217

Strüssmann CA, Yamamoto Y, Hattori RS, Fernandino JI, Somoza GM. Where the ends meet: an overview of sex determination in atheriniform fishes. Sexual Development. On-line. https://doi.org/10.1159/000515191

Castañeda-Cortés DC, Zhang J, Boan A, Langlois VS, Fernandino JI. High temperature stress response is not sexually dimorphic at the whole-body level and is dependent on androgens to induce sex reversal. General and Comparative Endocrinology. 299,113605. 2020. https://doi.org/10.1016/j.ygcen.2020.113605

Delgadin TH, Castañeda-Cortés DC, Sacks C, Breccia A, Fernandino JI, Vissio PG. Morphological colour adaptation during development: Involvement of Growth Hormone Receptor 1. Journal of Experimental Biology. 223. 2020. https://doi:10.1242/jeb.230375

González A, Kroll KJ, Silva-Sánchez C, Carriquiriborde P, Fernandino JI, Denslow ND, Somoza GM. Steroid hormones and estrogenic activity in the wastewater outfall and receiving waters of the Chascomús chained shallow lakes system (Argentina). Sci Total Environ 743: 140401. 2020.  https://doi.org/10.1016/j.scitotenv.2020.140401

Hattori RS, Castañeda-Cortés DC, Arias Padilla LF, Strobl-Mazzulla PH, Fernandino, JI.Activation of stress response axis as a key process in environment-induced sex plasticity in fish. Cell Mol Life Sci. 2020. https://doi.org/10.1007/s00018-020-03532-9

Lizarraga A, Klapholz O’Brown Z, Boulias K, Roach L, Greer E, Johnson PJ, Strobl-Mazzulla, PH, de Miguel N.Adenine DNA methylation, 3D genome organization and gene expression in the parasite Trichomonas vaginalis. P Natl Acad Sci USA. En prensa.

Castañeda Cortés DC, Arias Padilla LF, Langlois VS, Somoza GM, Fernandino JI. The central nervous system acts as a transducer of stress-induced masculinization through corticotropin-releasing hormone B. Development. 146: dev172866. 2019. https://doi.org/10.1242/dev.172866

Fernandino JI, Hattori RS. Sex determination in Neotropical fish: Implications ranging from aquaculture technology to ecological assessment. 2019. Gen Comp Endocrinol. 273: 172-183. 2019. https://doi.org/10.1016/j.ygcen.2018.07.002

González A, Fernandino JI, Elisio M, Chalde T, Miranda LA, Hammond GL, Somoza GM. Sex hormone binding globulin during an annual reproductive cycle in the hepatopancreas and ovary of pejerrey (Odontesthes bonariensis). Gen Comp Endocrinol. 272: 52-56. 2019. https://doi.org/10.1016/j.ygcen.2018.11.010

Hattori RS, Somoza GM, Fernandino JI, Colautti DC, Miyoshi K, Gong Z, Yamamoto Y, Strüssmann CA. The duplicated Y-specific amhy gene is conserved and linked to maleness in silversides of the genus Odontesthes. Genes (Basel). 10: 679. 2019. https://doi.org/10.3390/genes10090679

Sánchez-Vásquez E, Bronner ME, Strobl-Mazzulla PH. Epigenetic inactivation of miR-203 as a key step in neural crest epithelial-to-mesenchymal transition. Development. 146: dev171017. 2019. https://doi.org/10.1242/dev.171017