María Carmen Garza

Universidad de Zaragoza

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Juan José Badiola

Universidad de Zaragoza

Holger Wille

University of Alberta

Rosa Bolea

Universidad de Zaragoza

Eva Monleón

Universidad de Zaragoza

Cristina Acín

Universidad de Zaragoza

Belén Marín

Universidad de Zaragoza

Itziar Lamíquiz-Moneo

Hospital Universitario Miguel Servet

Leonardo M. Cortez

University of Alberta

Gültekin Tamgüney

Heinrich Heine University Düsseldorf

Judd M. Aiken

University of Alberta

Cooperative Institutions

Universidad de Zaragoza

Instituto de Investigación Sanitaria Aragón

Centro de Investigación y Tecnología Agroalimentaria de Aragón

University of Alberta

Instituto de Salud Carlos III

Centro de Investigación Biomédica en Red

Hospital Universitario Miguel Servet

Universitat Autònoma de Barcelona

German Center for Neurodegenerative Diseases

Women and Children’s Health Research Institute

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Influence of the different alleles of codon 222 of PRNP gene on susceptibility to bovine spongiform encephalopathy in goats.

XV Jornadas sobre Producción Animal, Zaragoza 14 y 15 de mayo de 2013 (2013)

José Luis Pitarch Jan Langeveld Alex Bossers Belén Marín F. Barillet

Bovine spongiform encephalopathy

PRNP

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Protein misfolding cyclic amplification corroborates the absence of PrP Sc accumulation in placenta from foetuses with the ARR/ARQ genotype in natural scrapie

Veterinary Microbiology (2017)

María Carmen Garza Hasier Eraña Joaquı́n Castilla Cristina Acín Antonia Vargas

10.1016/j.vetmic.2017.03.034

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Evaluation of ovine mesenchymal stem cells as in vitro models of scrapie.

XVIII Jornadas sobre Producción Animal, Zaragoza, España, 7 y 8 de mayo de 2019 (2019)

Adelaida Hernaiz L. Domínguez-Estallo S. Gómez-Redrado Óscar López-Pérez Mitch A. Garcia

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Toll‐like receptor‐mediated immune response inhibits prion propagation

Glia (2016)

Sang‐Gyun Kang Chiye Kim Leonardo M. Cortez María Carmen Garza Jing Yang

Prion diseases are progressive neurodegenerative disorders affecting humans and various mammals. The prominent neuropathological change in prion diseases is neuroinflammation characterized by activation of neuroglia surrounding prion deposition. The cause and effect of this cellular response, however, is unclear. We investigated innate immune defenses against prion infection using primary mixed neuronal and glial cultures. Conditional prion propagation occurred in glial cultures depending on their immune status. Preconditioning of the cells with the toll‐like receptor (TLR) ligand, lipopolysaccharide, resulted in a reduction in prion propagation, whereas suppression of the immune responses with the synthetic glucocorticoid, dexamethasone, increased prion propagation. In response to recombinant prion fibrils, glial cells up‐regulated TLRs (TLR1 and TLR2) expression and secreted cytokines (tumor necrosis factor‐α, interleukin‐1β, interleukin‐6, granulocyte‐macrophage colony‐stimulating factor, and interferon‐β). Preconditioning of neuronal and glial cultures with recombinant prion fibrils inhibited prion replication and altered microglial and astrocytic populations. Our results provide evidence that, in early stages of prion infection, glial cells respond to prion infection through TLR‐mediated innate immunity. GLIA 2016;64:937–951

Neuroglia

10.1002/glia.22973

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Bioluminescence Imaging of Neuroinflammation in Transgenic Mice After Peripheral Inoculation of Alpha-Synuclein Fibrils

Journal of Visualized Experiments (2017)

Sara Breid María E. Bernis Julius B. Tachu María Carmen Garza Holger Wille

To study the prion-like behavior of misfolded alpha-synuclein, mouse models are needed that allow fast and simple transmission of alpha-synuclein prionoids, which cause neuropathology within the central nervous system (CNS). Here we describe that intraglossal or intraperitoneal injection of alpha-synuclein fibrils into bigenic Tg(M83+/-:Gfap-luc+/-) mice, which overexpress human alpha-synuclein with the A53T mutation from the prion protein promoter and firefly luciferase from the promoter for glial fibrillary acidic protein (Gfap), is sufficient to induce neuropathologic disease. In comparison to homozygous Tg(M83+/+) mice that develop severe neurologic symptoms beginning at an age of 8 months, heterozygous Tg(M83+/-:Gfap-luc+/-) animals remain free of spontaneous disease until they reach an age of 22 months. Interestingly, injection of alpha-synuclein fibrils via the intraperitoneal route induced neurologic disease with paralysis in four of five Tg(M83+/-:Gfap-luc+/-) mice with a median incubation time of 229 ±17 days. Diseased animals showed severe deposits of phosphorylated alpha-synuclein in their brains and spinal cords. Accumulations of alpha-synuclein were sarkosyl-insoluble and colocalized with ubiquitin and p62, and were accompanied by an inflammatory response resulting in astrocytic gliosis and microgliosis. Surprisingly, inoculation of alpha-synuclein fibrils into the tongue was less effective in causing disease with only one of five injected animals showing alpha-synuclein pathology after 285 days. Our findings show that inoculation via the intraglossal route and more so via the intraperitoneal route is suitable to induce neurologic illness with relevant hallmarks of synucleinopathies in Tg(M83+/-:Gfap-luc+/-) mice. This provides a new model for studying prion-like pathogenesis induced by alpha-synuclein prionoids in greater detail.

Gliosis

Astrogliosis

Intraperitoneal injection

Synucleinopathies

10.3791/55503-v

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ADAPTACIÓN DE UN HALL ESCAPE PARA EL ESTUDIO DE LA ANATOMÍA HUMANA

Revista educación investigación innovación y transferencia (2022)

Alberto García‐Barrios Ana Isabel Cisneros Gimeno Itziar Lamíquiz-Moneo María Carmen Garza Jaime Whyte

Introducción: Las nuevas generaciones de alumnos, inmersas en un “mundo digital”, han promovido la aparición de nuevas metodologías docentes. En este trabajo, planteamos una serie de actividades a modo de Escape Room en la asignatura de Anatomía Humana II (Esplacnología) del Grado de Medicina de la Universidad de Zaragoza. Métodos: Se planteó una actividad Hall Escape Room basada en 5 retos secuenciales relacionados con los contenidos de la asignatura. La evaluación de la experiencia se planteó a los participantes mediante una encuesta de respuesta voluntaria de 5 preguntas basadas en la escala Likert (con 5 opciones de respuesta siendo 1 totalmente en desacuerdo y 5 totalmente de acuerdo). Resultados: La encuesta de valoración fue respondida por el 85 % del alumnado (76/89), aportando que estas actividades son positivas para la motivación, aprendizaje e integración de los contenidos teóricos y prácticas, así como fomentar la relación entre el alumnado. Conclusión: Las actividades de aprendizaje basado en juego tipo “Escape Room” mejoran la motivación y el aprendizaje del alumno, fomentando además la participación y comunicación entre ellos.

10.26754/ojs_reiit/eiit.202216761

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Detection of PrPres in Genetically Susceptible Fetuses from Sheep with Natural Scrapie

PLoS ONE (2011)

María Carmen Garza Natalia Fernández‐Borges Rosa Bolea Juan José Badiola Joaquı́n Castilla

Scrapie is a transmissible spongiform encephalopathy with a wide PrPres dissemination in many non-neural tissues and with high levels of transmissibility within susceptible populations. Mechanisms of transmission are incompletely understood. It is generally assumed that it is horizontally transmitted by direct contact between animals or indirectly through the environment, where scrapie can remain infectious for years. In contrast, in utero vertical transmission has never been demonstrated and has rarely been studied. Recently, the use of the protein misfolding cyclic amplification technique (PMCA) has allowed prion detection in various tissues and excretions in which PrPres levels have been undetectable by traditional assays. The main goal of this study was to detect PrPres in fetal tissues and the amniotic fluid from natural scrapie infected ewes using the PMCA technique. Six fetuses from three infected pregnant ewes in an advanced clinical stage of the disease were included in the study. From each fetus, amniotic fluid, brain, spleen, ileo-cecal valve and retropharyngeal lymph node samples were collected and analyzed using Western blotting and PMCA. Although all samples were negative using Western blotting, PrPres was detected after in vitro amplification. Our results represent the first time the biochemical detection of prions in fetal tissues, suggesting that the in utero transmission of scrapie in natural infected sheep might be possible.

Transmissible spongiform encephalopathy

10.1371/journal.pone.0027525

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Fibronectin: a natural barrier against prion infection

bioRxiv (Cold Spring Harbor Laboratory) (2023)

María Carmen Garza Sang Gyun Kang Chiye Kim Eva Monleón Jacques van der Merwe

Abstract A distinctive signature of the prion diseases is the accumulation of the pathogenic isoform of the prion protein, PrP Sc , in the central nervous system of prion-affected humans and animals. PrP Sc is also found in peripheral tissues, raising concerns about the potential transmission of pathogenic prions through human food supplies and posing a significant risk to public health. Although muscle tissues are considered to contain levels of low prion infectivity, it has been shown that myotubes in culture efficiently propagate PrP Sc . Given the high consumption of muscle tissue, it is important to understand what factors could influence the establishment of a prion infection in muscle tissue. Here we used in vitro myotube cultures, differentiated from the C2C12 myoblast cell line (dC2C12), to identify factors affecting prion replication. A range of experimental conditions revealed that PrP Sc is tightly associated with proteins found in the systemic extracellular matrix (ECM), mostly fibronectin (FN). The interaction of PrP Sc with FN decreased prion infectivity, as determined by standard scrapie cell assay. Interestingly, the prion-resistant reserve cells in dC2C12 cultures displayed a FN-rich ECM while the prion-susceptible myotubes expressed FN at a low level. In agreement with the in vitro results, immunohistopathological analyses of tissues from sheep infected with natural scrapie demonstrated a prion susceptibility phenotype linked to an extracellular matrix with undetectable levels of FN. Conversely, PrP Sc deposits were not observed in tissues expressing FN. These data indicate that extracellular FN may act as a natural barrier against prion replication and that the extracellular matrix composition may be a crucial feature determining prion tropism in different tissues. Author summary Prion diseases are complex fatal neurodegenerative disorders caused by a misfolded form of the cellular protein PrP C (PrP Sc ). Due to the potential zoonotic transmission of these disorders through animal-based food intake, it is crucial to identify the tissues in which PrP Sc can accumulate and what might influence its tropism. Animal muscle (or meat) and related food products are highly consumed, raising concern of the involvement of muscle cells in prion replication. Muscle tissue from prion-affected animals contains low levels of infectivity and PrP Sc is mostly associated with nerve structures rather than myofibers, whereas C2C12 myotubes, a muscle-derived cell type, efficiently replicate prions in vitro and generate high levels of infectivity compared with other cell cultures. We demonstrate a fibronectin-mediated interference with prion infection in differentiated C2C12 cultures that correlates with the findings in tissues from naturally scrapie-infected animals. Our results suggest that extracellular matrix composition, specifically regarding the presence of fibronectin, might determine prion tropism and dissemination.

C2C12

Infectivity

10.1101/2023.08.31.555658

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Online Teaching Alternative in Human Anatomy

Anatomia (2022)

Alberto García‐Barrios Ana Isabel Cisneros Gimeno María Carmen Garza Itziar Lamíquiz-Moneo Jaime Whyte

The objective was to implement a “Breakout” activity using an online platform (Genially®) similar to those carried out in person to increase interactivity, motivation, and teamwork. The activity was proposed during the 2021–2022 academic year in the Human Anatomy II (Splanchnology) course taken in the second semester of the Bachelor’s Degree in Medicine (University of Zaragoza, Zaragoza, Spain) and was carried out with the participation of 89 students enrolled in the course. The evaluation of the experience by the students was carried out by means of an online questionnaire that comprised four questions (based on the Likert scale) and by the teaching staff through a coordination meeting. In total, 86% of the students agreed regarding the positive effect of these kinds of activities on interactivity and motivation, with 65% agreeing on the usefulness of these tools. Around 70% agree that this activity helped them to integrate course content and to enhance teamwork.

Interactivity

Bachelor

Human anatomy

Computer-assisted web interviewing

10.3390/anatomia1010009

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Detection of PrPSc in the Skin of Sheep Infected Naturally by Scrapie

Journal of Comparative Pathology (2013)

María Carmen Garza Cristina Acín Marta Monzón Rosa Bolea Juan José Badiola

10.1016/j.jcpa.2012.11.109

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