VISEGRÁD GROUP SOCIETY FOR DEVELOPMENTAL BIOLOGY - V4SDB 2020
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2 n d CONFERENCE OF THE
VISEGR ÁD GROUP SOCIE T Y
FOR DE VELOPMENTAL BIOLOGY
2 – 5 September, 2021, Szeged, Hungary
IN THE ORGANIZ ATION OF THE V4 SOCIET Y FOR DEVELOPMENTAL
BIOLOGY (V4SDB) AND THE HUNGARIAN GENETICS SOCIET Y (MAGE)
ABSTR AC T S
OR AL AND POSTER PRESENTATIONS
ABSTRACTS | ORAL PRESENTATIONS 3
ABSTRACTS chromatin remodelling. In most animal, a characteristic feature of
this change is the replacement oocyte/early embryo specific linker
ORAL PRESENTATIONS histone by somatic histone H1 variants. To uncover the specific role
of this alternative linker histone in early Drosophila embryogenesis,
we established fly lines in which domains of embryo specific linker
histone, BigH1 have been replaced partially or completely with that
T1. KEYNOTE TALK 1 of somatic H1. Analysis of this mutant Drosophila lines revealed that
Cytoplasmic forces functionally reorganize nuclear conden- H1 can substitute BigH1 under normal conditions, however at low
sates in oocytes temperature, globular and C-terminal domains of BigH1 are nec-
Marie-Hélène Verlhac 1 essary for proper development. We showed that in the presence
1
Center for Interdisciplinary Research in Biology, College de France, of BigH1, nucleosome stability increased compared to H1 however
Paris, France core histone exchange during replication is more dynamic. Based on
our results we propose a model explaining how BigH1 ensures fast
Abstract was not received. paced replication cycles before zygotic genome activation.
T1. KEYNOTE TALK 2 T4.
Understanding changing architecture of the mammalian p38-MAPK-mediated translation regulation primes blastocyst
embryo development and is required for primitive endoderm differen-
Berenika Plusa 1 tiation in mice
1
University of Manchester, Division of Developmental Biology and Pablo Bora 1, Lenka Gahurova 1,2, Tomáš Mašek 3,
Medicine, United Kingdom Andrea Hauserova 1, David Potěšil 4, Denisa Jansova 2, Andrej Susor 2,
During mammalian preimplantation development, cells reduce in Zbyněk Zdráhal 4, Anna Ajduk 5, Martin Pospíšek 3,
size and the architecture of the embryo changes significantly. Such Alexander W. Bruce 1
changes are accompanied by dynamic adjustments in gene expres- 1
Laboratory of Early Mammalian Developmental Biology (LEMDB),
sion and specification of the first three embryonic lineages. Two of Department of Molecular Biology & Genetics, Faculty of Science, Univer-
them, the trophectoderm (TE) and the primitive endoderm (PrE) sity of South Bohemia, Branišovská 31, 37005 České Budějovice, Czech
will give rise to extraembryonic tissues such as placenta and part Republic; 2 Laboratory of Biochemistry and Molecular Biology of Germ
of the yolk sac respectively. At the same time, a group of cells, the Cells, Institute of Animal Physiology and Genetics, CAS, Rumburská
epiblast (Epi), needs to be protected from the differentiation signals 89, 27721 Liběchov, Czech Republic; 3 Laboratory of RNA Biochemistry,
to assure development of the foetus. Dynamic crosstalk between Department of Genetics and Microbiology, Faculty of Science, Charles
the gene regulatory networks and the evolving architecture of the University, Viničná 5, 12844 Prague 2, Czech Republic; 4 Central Euro-
mammalian embryo are notoriously difficult to study and currently, pean Institute of Technology, Masaryk University, 62500 Brno, Czech
no easily accessible open-source tool exists that would allow for Republic; 5 Department of Embryology, Institute of Developmental Biology
large-scale, quantitative assessment of the changes in the cell and Biomedical Sciences, Faculty of Biology, University of Warsaw, Miec-
microenvironment. With this in mind, we developed the quantifi- znikowa 1, 02-096, Warsaw, Poland
cation pipeline IVEN (Internal Versus External Neighbourhood), an
Keywords: mouse blastocyst, primitive endoderm differentiation,
open-source and user-friendly software that can be used in con-
regulation of translation
junction with existing open-source image analysis software like
ImageJ, as well as commercially available programs like IMARIS. Successful specification of the two mouse blastocyst inner cell mass
With the use of IVEN, we studied the changes in cell neighbour- (ICM) lineages (the primitive endoderm (PrE) and epiblast) is a pre-
hood architecture that accompany self-organisation and lineage requisite for continued development and requires active fibroblast
formation in the mammalian embryo and revealed how modulation growth factor 4 (FGF4) signaling. Previously, we identified a role
of signalling pathways can alter embryo geometry. for p38-mitogen-activated protein kinases (p38-MAPKs) during
PrE differentiation, but the underlying mechanisms have remained
T3. unresolved. Here, we report an early blastocyst window of p38-
Is bigger also better? Functional comparison of somatic and MAPK activity that is required to regulate ribosome-related gene
larger, embryo specific linker histone during early Drosophila expression, rRNA precursor processing, polysome formation and
embryogenesis. protein translation. We show that p38-MAPK inhibition-induced
László Henn 1, 2, Anikó Szabó 1,3, Balázs Vedelek 3, Imre M. Boros 1, 3 PrE phenotypes can be partially rescued by activating the transla-
1
Institute of Biochemistry, Biological Research Centre, Szeged, Hungary; tional regulator mTOR. However, similar PrE phenotypes associated
2
Institute of Genetics, Biological Research Centre, Szeged, Hungary; with extracellular signal-regulated kinase (ERK) pathway inhibition
3
Department of Biochemistry and Molecular Biology, University of targeting active FGF4 signalling are not affected by mTOR acti-
Szeged, Hungary vation. Moreover, we identify and verify translation related can-
didate p38-MAPK effectors/ substrates and confirm their role in
Keyworlds: Chromatin, linker histone, embryogenesis
blastocyst maturation. These data indicate a specific role for p38-
The earliest steps of animal development are driven by maternally MAPKs in providing a permissive translational environment during
deposited gene products since transcription program is still inac- mouse blastocyst PrE differentiation that is distinct from classically
tive. The zygotic genome activation is accompanied by a general reported FGF4-based mechanisms.
4 ABSTRACTS | ORAL PRESENTATIONS
T5. and cell fate of rabbit isolated ICMs. Our study points to significant
The detachment of the blastoderm-vitelline envelope interac- inter-species differences, most notably en extended period of ICM
tion and blastoderm chirality potency to differentiate into TE lineage.
Giulia Serafini 1, Marina Cuenca 1, Pavel Tomančak 1,2 We first identify GATA3 as an early marker of rabbit TE and CDX2
1
Max Planck Institute of Molecular Cell Biology and Genetics – as a marker of more mature TE, as CDX2 expression is initiated at
MPI-CBG, Dresden, Germany; 2 Central European Institute of Technology mid-blastocyst stage. We then analyse developmental potential
– CEITEC, Brno, Czech Republic of rabbit ICMs isolated by immunosurgery and subsequently cul-
tured in vitro. ICMs originating from early- to mid-blastocyst stage
Keywords: gastrulation, attachment, embryonic chirality embryos are able to re-form a blastocyst-like structure, with a func-
Gastrulation is a complex and well-coordinated process that, tional TE, and an ICM containing both SOX2-positive epiblast cells
through a precise combination of tissue rearrangements and cel- and SOX17-positive primitive endoderm cells. We further observe
lular migrations, leads to the segregation of the germ layers in the that rabbit ICMs isolated form later blastocyst stages lose the ability
developing embryo. Its principal issue is movement: both single cells for TE specification. Instead, these ICMs form a halo-like cavity with
and tissues change dramatically their relative positions over time, an outer layer of SOX17-positive cells, indicating that the potential
being strongly influenced by the interaction with their surround- for TE differentiation gives way to formation of a different type of
ings. Of particular interest to us is the role played by the vitelline epithelial extraembryonic layer.
envelope, the innermost layer of the eggshell. Our group recently
described that integrin-mediated attachment of the blastoderm T7.
to the vitelline envelope is required for the proper gastrulation of The role of activin A in the regulation of preimplantation
both Drosophila melanogaster and Tribolium castaneum. The dis- development of mouse embryo
ruption of the attachment dramatically alters gastrulation in both Eliza Winek 1,2, Katarzyna Szczepańska 1, Aneta Suwińska 1
organisms and in Drosophila the defect is scoreable as a twisted 1
Department of Embryology, Institute of Developmental Biology and Bio-
gastrulation (TG) phenotype. Exploiting this, we searched for addi- medical Research, Faculty of Biology, University of Warsaw, Warsaw,
tional molecular players involved in the attachment besides integ- Poland; 2 Doctoral School of Exact and Natural Sciences, Warsaw, Poland
rins. Surprisingly, when quantifying the TG phenotype, we detected
Keywords: activin A, mouse, blastocyst, cell lineages
a bias in the handedness of the twist. This led us to hypothesize
that the attachment could have the role of keeping the embryo Activin A, encoded by InhbA gene, is a protein belonging to the
symmetric, preventing the germ band from following the intrinsic transforming growth factor β (TGFβ) family, which is known for its
chirality of the tissue during germ band extension. Since the ear- involvement in the patterning of the embryo after implantation.
liest developmental process showing chirality is gut formation, our Although the expression of activin A is also detected in the preim-
results suggest that the left/right asymmetry could be established plantation stages of mouse (and human) embryo development, its
much earlier, during gastrulation. Moreover, the deletion screening role in this period of embryogenesis remains undiscovered.
on mutants showing the TG phenotype provided a few interesting We aimed to determine whether activin A pathway is engaged
candidate genes, which will allow us to look at left/right asymme- in the commitment of primary cell lineages in the mouse pre-
try establishment in Drosophila embryo from a novel and surprising implantation embryo. To this end, we analyzed development of
point of view. activin A-deficient mouse embryos (InhbA-/-) from the zygote to the
blastocyst stage using time-lapse microscopy and compared them
T6. with the stage-matched heterozygotes and wild-type embryos. We
Rabbit inner cell mass retains potential to differentiate into the revealed that the absence of zygotic activin A did not affect the rate
trophectoderm lineage of development of InhbA-/- embryos (pronuclear envelope break-
Katarzyna Filimonow 1, Anna Chołoniewska 1, Jan Chołoniewski 2, down, cleavages, compaction, cavitation, hatching, duration of cell
Zofia Eliza Madeja 3, Katarzyna Barłowska 1, Elżbieta Wenta- cycles and synchrony of cleavage rounds). However, we observed
Muchalska 1, Berenika Plusa 4, Anna Piliszek 1 that the absence of zygotic activin A disrupts the correct propor-
1
Department of Experimental Embryology, Institute of Genetics and Ani- tion of cells within the inner cell mass (ICM), increases the partici-
mal Biotechnology of the Polish Academy of Sciences, Jastrzebiec, Poland; pation of epiblast (EPI) at the expense of primitive endoderm (PE)
2
Center of Excellence for Complex Systems Research, Faculty of Physics, cells. Thus, our preliminary results indicate that activin A may be
Warsaw University of Technology, Warsaw, Poland; 3 Faculty of Veter- engaged in establishment of the final EPI:PE ratio during preimplan-
inary Medicine and Animal Sciences, Poznan University of Life Sciences, tation period of mouse development.
Poznan, Poland; 4 Division of Developmental Biology & Medicine, Univer- Since InhbA-/- embryos were able to reach the blastocyst stage, our
sity of Manchester, Manchester, United Kingdom next goal is to verify whether maternal activin A can partially com-
pensate for the loss of zygotic gene function.
Keywords: lineage specification, trophectoderm, rabbit Acknowledgement: The project is financed by the grant OPUS 17
In the course of mammalian development initial state of totipo- 2019/33/B/NZ3/02906 from the National Science Centre (Poland).
tency has to be lost to allow acquisition of a specific cell fate. The
first differentiation event results in the formation of trophectoderm
(TE) and the inner cell mass (ICM). In the mouse embryo the cell
fate of these two compartments is set quickly after formation of
a blastocyst. Here we present a detailed analysis of development
ABSTRACTS | ORAL PRESENTATIONS 5
T8. KEYNOTE TALK 3 T10.
Keynote lecture: Aberrant activation of developmental pro- Unique stem cell subpopulation which ensures mesenchymal
grams in adult disease regeneration of continuously growing teeth contributes to
Ángela Nieto various tissue organogenesis
Instituto de Neurociencias (CSIC-UMH), Alicante, Spain Jan Krivanek 1, Josef Lavicky 1, Petr Taus 2, Marcos Gonzalez Lopez 1,
Vladislav Rakultsev 1, Tereza Kurucova 2, Aneta Anna Dunajova 1,
Epithelial homeostasis is crucial to maintain tissue architecture, and
Marcela Buchtova 3,4, Marie Sulcova 3,4
therefore, it needs to be tightly regulated in the adult. By contrast, 1
Department of Histology and Embryology, Faculty of Medicine, Masaryk
embryonic cells show a high degree of epithelial plasticity required
University, Brno, Czech Republic; 2 Central European Institute of Technol-
for proper morphogenesis and, in particular, for the implementation
ogy, Masaryk University, Brno, Czech Republic; 3 L aboratory of Molecular
of massive cell movements that occur during gastrulation and neu-
Morphogenesis, Institute of Animal Physiology and Genetics, Czech Acad-
ral crest delamination among other processes. We have been inter-
emy of Science, Brno, Czech Republic; 4 Department of Experimental Biol-
ested in the analysis of cell movements, plasticity and epithelial to
ogy, Faculty of Science, Masaryk University, Brno, Czech Republic
mesenchymal transitions (EMT) for many years, and found that the
aberrant activation of developmental EMT-like programs in adult Keywords: Stem cells, development, odontogenesis, regeneration
cells leads to several pathologies including tumor progression and
In rodents and several other species, a continuously growing teeth
organ degeneration. While the epithelial and mesenchymal cells are
have evolved. This adaptation on specialized style of living ensures
usually considered as extreme phenotypes, intermediate EMT states
permanent replenishing of dental tissues worn by constant gnawing
also exist. Under those circumstances cells depict a hybrid pheno-
and provides an attractive system for studying of stem cell niche,
type expressing both epithelial and mesenchymal markers and from
cell differentiation or injury-induced regeneration. Recent advances
which they can reverse to the original state or move towards a more
in single cells RNA sequencing and lineage tracing methods enabled
mesenchymal phenotype. Hybrid transitory states can favor coordi-
to perform an unbiased and reliable analysis of this organ and to
nated cell migration or wound healing but they can also enable the
study different stem cell populations responsible for permanent
formation of clusters of migratory cancer cells with increased met-
growth.
astatic potential. However, in contrast to the situation in cancer, the
Using this approach, we found a novel, quiescent and long-lasting
intermediate phenotype holds promise for new antifibrotic ther-
population of mesenchymal stem cells contributing to the perma-
apeutic approaches, as inhibiting EMT can attenuate established
nent tooth growth. This, up to now unknown, stem cell population
fibrosis. I will discuss different scenarios in which this intermediate
is spatially restricted and gives rise to all mesenchymal parts of
phenotype is observed both in development and in disease.
dental pulp, including different types of dental pulp cells and den-
tin-producing odontoblasts. Further analyses showed a multipotent
T9.
characteristic of this unique population and uncovered molecular
Pharyngeal component in the premandibular segment of the background responsible for differentiation (bifurcations) into dis-
vertebrate head tinct terminally differentiated cell states. Based on our detailed
Horackova, A., Stundl, J., Minarik, M., Fabian, P. & Cerny, R. study of this exemplary model system on single cell level we
Dept. Zoology, Charles University in Prague uncovered the role of the same type of stem cell population during
The segmental formation of the pharynx represents a fundamental embryonic development of several organ systems.
part of the metameric organization of the vertebrate head and face. Taken together we discovered a novel, highly specific mesenchymal
Pharyngeal pouches maintain populations of the craniofacial mes- stem cells which plays role during permanent adult tissue growth
enchyme separate, and signalling from the endodermal pouches is and contribute to formation of different organs during development.
essential for induction and pattern of serially arranged cranial skel-
etal elements. Segmentation of the vertebrate head is thus orches- T11.
trated around pouching of the primitive gut cavity, which forward Developmental anoikis: A novel, protective safeguarding
expansion is thought to be limited by the oral (stomodeal) invag- mechanism in the embryonic neocortex
ination during the mouth formation. Recently, we ascertained the Zsolt Lele 1*, Zsófia I. László 1*, Fruzsina Mógor 1, Dániel Nagy 1,
existence of the preoral gut (POG), the rostral-most foregut pouch Dárius Leszkó 1, Susanne Prokop 1, Gabriel M. Simon 2, Ken Mackie 3,
located dorsoanterior to the forming mouth, and thus extending to Benjamin F. Cravatt 2 and István Katona 1,3
the pre mandibular segment. POG represents a classical, yet forgot- *=contributed equally
ten textbook embryonic domain, and we argue that it presents a 1
Institute of Experimental Medicine, Hungarian Academy of Science, Buda-
deep deuterostome heritage of the pan-vertebrate pharynx. Devel- pest, Hungary; 2 Department of Chemical Physiology, The Scripps Research
opmental and evolutionary significance of the pharyngeal pouch- Institute, La Jolla, California, USA; 3 Gill Institute and Department of
like component in the rostral-most, premandibular head segment Psychological and Brain Sciences, Indiana University, Bloomington, Indi-
will be discussed in the context of the segmental nature of the ver- ana, USA
tebrate head.
Proliferating progenitor cells in higher organisms are generally
located in designated stem cell niches where they are immobilized
via cell-to-cell and cell-to-ECM connections. In the embryonic cor-
tex the ventricular zone (VZ) is such a neurogenic niche occupied
mostly by modified neuroepithelial cells called radial glia progen-
6 ABSTRACTS | ORAL PRESENTATIONS
itor cells (RGPCs). Asymmetric division of RGPCs produce a radial T13.
glia cell and an intermediate progenitor cell (IPCs) or fate-comit- Shedding light on the details of hemocyte transdifferentia-
ted neuroblast which migrate radially to their proper destination. tion in Drosophila melanogaster with an artificial intelligence
To escape the VZ, these cells need to loose their cadherin-based based technology
adherens junctions to neighbor cells at the apical, and laminin-inte- Erika Gábor 1, Aliz Géczi 1, Ede Migh 2, Attila Beleon 2,
grin based cell-ECM connections at the pial surface. This process is Péter Horváth 2, Bence Széplaki 3, Márton Enyedi 3, Lajos Pintér 3,
largely reminiscent of the classic epithelial-mesenchymal transition Lajos Haracska 3, Viktor Honti 1
(EMT). EMT is essential during certain steps of development (gas- 1
Drosophila Blood Cell Differentiation Group, Genetic Institute, ELKH,
trulation, neural crest) but can be a harmful process in the adult Szeged, Hungary; 2 Laboratory of Mutagenesis and Carcinogenesis
organism (fibrosis, metastasis). To prevent unwanted migration Research, Genetic Institute, ELKH, Szeged, Hungary; 3 Laboratory of
of cells in adult epithelial tissues, loss of cell to ECM connections Microscopic Image Analysis and Machine Learning, Genetic Institute,
usually evokes a specific apoptotic process called anoikis. In this ELKH, Szeged, Hungary
talk I’ll present data from a project in which we sought to establish
whether there is a similar protective cell death process in the devel- Keywords: Drosophila, hemocyte, differentiation, transdifferentia-
oping brain and share some details about the molecular machinery tion, immunity
behind this phenomenon. Organisms are highly adaptive to the ever-changing environment.
To establish a fast and efficient response, reusing the already active
T12. energy sources is a requirement. A good example for this phenom-
Primary Cilia and Kidney Cysts Development in Nek8-deficient enon is transdifferentiation, during which a functional cell further
Mice differentiates into a functionally distinct one without the involve-
Kristekova Daniela 1,2, Buchtova Marcela 1,2 ment of stem cells. Both in vertebrates and invertebrates, blood
1
Laboratory of Molecular Morphogenesis, Institute of Animal Physiol- cells are capable for transdifferentiation under certain conditions.
ogy and Genetics, v.v.i., Czech Academy of Sciences, Brno 602 00, Czech For detailed investigation of transdifferentiation, the fruit fly (Dro-
Republic; 2 Section of Animal Physiology and Immunology, Department of sophila melanogaster) is a perfect model: the vast majority of its
Experimental Biology, Faculty of Science, Masaryk University, Brno 625 signaling pathways, transcription- and epigenetic factors regulating
00, Czech Republic hematopoiesis are evolutionary conserved, and its hemocytes show
remarkable similarities in their function and differentiation to the
Keywords: primary cilia, kidney, cyst formation
vertebrate myeloid cells.
Polycystic kidney diseases are a family of genetic disorders man- Previously we discovered that the phagocytic plasmatocytes
ifested by renal cystic growth, resulting in kidney failure. Mouse are capable of converting into encapsulating lamellocytes upon
line C57BL/6J-Nek8jck/J carries the Nek8 mutation, which in reces- immune induction (e.g. wasp infestation, wounding). The latest sin-
sive homozygotes leads to juvenile cystic kidneys-like phenotype. gle-cell transcriptome data proved that plasmatocytes - which were
The first small cysts, mostly located in the kidney cortex, can be believed to be homogeneous - form several functional clusters.
observed at 4 weeks with typical progressive growth, which can We established a novel method to explore the plasmatocyte-lamel-
be observed with the age of animals. The lifespan of the Nek8-mu- locyte transdifferentiation. In an ex vivo culture, we identify and
tant mice is around 20 – 25 weeks. Nek8-mutation was previously follow hemocytes by their cell-type specific transgenic fluorescent
associated with longer cilia and the accumulation of polycystin-1 reporter signal and take time-lapse images, which are analyzed with
and polycystin-21 in the cilia. The dysfunctional serine/threonine an artificial intelligence-based machine learning technology.
kinase Nek8 leads to disruption of cystogenesis by altering the We would like to shed light on the details of transdifferentiation
cilia morphology and function in the distal tubule of nephrons. In upon immune induction, as well as in tumorous mutants. Due to
our study, we focus on the changes in the expression of the genes evolutionary conservation, our results can open up new fields in the
associated with FGF/FGFR signalling and SHH pathway during the investigation of mammalian hematopoiesis and blood cell originated
development of the cysts with the aim to reconstruct the aber- cancer research.
rant processes in Nek8-mutants leading to kidney failure and their
association to primary cilia function failure. Kidneys from various T14. KEYNOTE TALK 4
stages of both mutant and wild-type mice were collected, analyzed Posttranscriptional control of the germline-soma dichotomy
for histopathological changes and alterations in gene expression. Rafal Ciosk
Moreover, we examined the possible curative effect of FGFR inhib- Dept. of Biosciences, University of Oslo, Norway
itor, Derazantinib (ARQ-087) on cyst formation in Nek8-deficient
mice. In summary, our study can provide us with deeper insight into Cell fate commitment and reprogramming are fundamental for
molecular regulations leading to the cyst formation caused by the development and tissue homeostasis. A profound cellular repro-
disruption of primary cilia and also lead to the design of new treat- gramming occurs during the oocyte-to-embryo transition, when
ments, which effect can be easily tested on the postnatal kidney specialized reproductive cells - egg and sperm - give rise to a totipo-
cysts development in our model. tent embryo. The majority of reported “roadblocks” controlling this
Acknowledgement: This work was supported by the Ministry of reprogramming are transcription factors and chromatin modifiers.
Health (NU20-08-00205). However, we demonstrated that RNA-binding proteins (RBPs) fulfill
a similar role in the oocyte cytoplasm. Using the C. elegans model,
we showed that conserved RBPs function as components of a
ABSTRACTS | ORAL PRESENTATIONS 7
posttranscriptional network that prevents oocytes from precocious T16.
reprogramming into embryonic-like cells. In the absence of this layer Golden hamster piRNAs are necessary for zygote development
of regulation, oocytes prematurely undergo embryonic-like differ- and establishment of spermatogonia
entiation and form teratomas. We are interested in understanding Zuzana Loubalova 1†, Helena Fulka 1,†‡, Filip Horvat 1,2,
the underlying molecular mechanisms, the connection between Josef Pasulka 1, Radek Malik 1, Michiko Hirose 3, Atsuo Ogura 3,4,
cytoplasmic and nuclear processes, and the coordination between Petr Svoboda 1
cellular reprogramming and other developmental events. 1
Institute of Molecular Genetics of the Czech Academy of Sciences,
Videnska 1083, 142 20 Prague 4, Czech Republic; 2 Bioinformatics Group,
T15. Division of Molecular Biology, Department of Biology, Faculty of Science,
Cytoplasmic RNA exosome is essential for post-implantation University of Zagreb, 10000, Zagreb, Croatia; 3 Bioresource Engineering
mouse embryo development Division, RIKEN BioResource Research Center, 305-0074 Ibaraki, Japan;
Michał Brouze 1, Marcin Szpila 1, Tomasz Kuliński 1, Wiktor Antczak 2, 4
Bioresource Engineering Laboratory, RIKEN Cluster for Pioneering
Areta Czerwińska 4, Jakub Gruchota 1, Ewa Borsuk 1,3, Research, 351-0198 Saitama, Japan.
Andrzej Dziembowski 1,2 † These authors contributed equally
1
Laboratory of RNA Biology, International Institute of Molecular and ‡ Current address: Institute of Experimental Medicine of the Czech
Cell Biology, Warsaw, Poland; 2 Laboratory of RNA Biology, Institute of Academy of Sciences, Videnska 1083, 142 20 Prague 4, Czech Republic
Genetics and Biotechnology, University of Warsaw, Poland; 3 Department
Keywords: piRNA, PIWI, small RNAs, Syrian hamster, retrotrans-
of Developmental Biology, Institute of Developmental Biology and Bio-
posons, oogenesis, spermatogenesis
medical Sciences, Faculty of Biology, University of Warsaw; 4 Department
of Cytology, Institute of Developmental Biology and Biomedical Sciences, PIWI-associated RNAs (piRNAs) maintain the integrity of the ger-
Faculty of Biology, University of Warsaw, Poland mline genome by suppressing retrotransposon activity, and their
functions in gene regulation have recently emerged. The under-
Keywords: mouse development, RNA, exosome
standing of the role of piRNA pathway in mammals was shaped by a
RNA exosome is an evolutionary conserved ribonucleolytic complex. mouse model in which piRNAs are essential for male but not female
In mammals, cytoplasmic and nuclear exosomes have a non-over- fertility. We report that the role of the piRNA pathway substan-
lapping set of catalytic subunits. Nuclear physiological processes tially differs in golden hamsters, whose piRNA pathway setup more
such as degradation of pervasive transcription products are medi- closely resembles that of other mammals, including humans. We
ated by DIS3 and EXOSC10. The role of the cytoplasmic exosome, show that the loss of Mov10l1 helicase, an essential factor initiating
equipped with DIS3L as a catalytic subunit, is less clear. piRNA biogenesis, generates novel phenotypes in hamsters. Unlike
As a part of our effort to study this role, we used CRISPR/Cas9 mice, female Mov10l1–/– hamsters are sterile. Mutant fully-grown
technology to create a transgenic mouse line with knock-out (KO) oocytes show minor changes in the transcriptome and moderate
mutation in Dis3l. This mutation causes embryolethality between to low increase in retrotransposon transcripts. Although Mov10l1–/–
day 6,5 and 7,5 of embryo development, with the ratio of KO to hamster oocytes appear largely normal and give rise to zygotes,
wild-type embryos being reduced after day 4,5. At blastocyst stage they cannot support normal embryonic development. In another
KO embryo growth rate is not impaired, but the development of contrast to mice, Mov10l1–/– male hamsters have impaired estab-
particular cell lineages is affected as indicated by changed cell num- lishment of spermatogonia accompanied by transcriptome dysregu-
ber of trophectoderm and primitive endoderm as well as the low lation and a surge of expression of a specific young retrotransposon
efficiency with which KO blastocyst produce embryonic stem cells subfamily, which is the primary target of retrotransposon-derived
(ES cells). Supplementation of 4-cell KO embryos with tetraploid piRNAs in early spermatogenesis. In addition, we examined groups
wild-type embryos didn’t produce any offspring, pointing to the of piRNAs during different stages of spermatogenesis and demon-
issue with inner cell mass as a source of embryolethality. strate that the regulation of specific retrotransposon subfamilies
While sequencing of RNA from KO ES cell lines suggested downreg- is stage-dependent. Our results not only refute the notion that in
ulation of 21 transcripts, RNA sequencing of single KO blastocysts mammals the piRNA pathway is important only in males but also
revealed the opposite trend of 11 deregulated transcripts out of demonstrate the adaptive nature of the mammalian piRNA path-
which 9 were upregulated. way, which allows to confront emerging genomic threats and
Taken together, these results allow us to describe the role of DIS3L acquire new critical roles in both sexes.
and cytoplasmic exosome in the development of mouse embryo
and its specific cell lineages, as well as identify transcripts regulated T17.
by DIS3L activity in developing embryo. The snoRNAome of zebrafish (Danio rerio)
Renáta Hamar and Máté Varga
Department of Genetics, ELTE Eötvös Loránd University, Budapest, Hungary
Small nucleolar RNAs (snoRNAs) are one of the most abundant
and evolutionary ancient group of functional non-coding RNAs.
They were originally described as guides of post-transcriptional
rRNA modifications, but snoRNAs fulfill an impressive variety of
cellular functions. These range from guiding site-specific chemical
modifications in several RNA classes and affecting the nucleolytic
8 ABSTRACTS | ORAL PRESENTATIONS
processing of ribosomal RNA (rRNA) to their involvement in the reg- Biologically, the most meaningful way to register the images is by
ulation of chromatin architecture and alternative splicing. Previous identifying cellular correspondences between these two imaging
analysis suggests that snoRNAs and the modifications they mediate modalities. In this way, one can bring the two sources of infor-
are highly conserved across species. mation into a single domain and combine dynamic information on
Based on common sequence motifs and structural features snoRNAs morphogenesis with static gene expression data. Here we propose
are classified in two major families, box C/D and box H/ACA snoR- a new computational pipeline for identifying cell-to-cell correspon-
NAs. The identification of snoRNAs based on homology search is dences between images from multiple modalities and for using
often difficult due to the lack of overall sequence conservation, small these correspondences to register 3D images within and across
size (60-300bp) and short sequence motifs. Here, we use size-frac- imaging modalities. We demonstrate this pipeline by combining
tionated RNA sequencing data from adult zebrafish tissues to define four-dimensional recording of embryogenesis of Spiralian annelid
the snoRNAome for this species. Our approach allowed us to identify ragworm Platynereis dumerilii with three-dimensional scans of fixed
several hitherto unannotated snoRNAs in the zebrafish genome. Platynereis dumerilii embryos stained for the expression of a variety
We created an analysis pipeline where the raw reads were aligned of important developmental genes. The complete pipeline is avail-
to the latest zebrafish genome assembly (GRCz11) using Bowtie2 able for public use through a napari plugin.
aligner and we identified putative snoRNA sequences using the
blockbuster algorithm. Using several snoRNA predictor methods we T19.
were able to confirm the presence of multiple previously predicted How to choose the most suitable increased fidelity SpCas9
snoRNAs in the data and also identified ~80 new snoRNA-like nuclease for high specificity genome editing
sequences missing from the current Ensembl database (v99). Based Péter István Kulcsár 1,2, András Tálas 1,3, Sarah L Krausz 1,3,
on our preliminary survey our snoRNAome dataset represents the Zoltán Ligeti 1,4, Vanessza L Végi 1, Eszter Tóth 1, Zsófia Rakvács 1,
most reliable set of snoRNAs to date in this species. Krisztina Huszár 1, Ervin Welker 1,5
We also compared all putative, novel snoRNAs with ncRNA sets 1
Research Centre for Natural Sciences, Institute of Enzymology, Budapest,
from zebrafish and other species via RNAcentral to identify already Hungary; 2 Department of Biophysics and Radiation Biology, Semmelweis
annotated zebrafish snoRNAs. We noted that four of these were University, Budapest, Hungary; 3 Semmelweis University, School of Ph.D.
already included in some (non-Ensemble) databases. We also noted Studies, Budapest, Hungary; 4 University of Szeged, Szeged, Hungary;
that another six of them were in zebrafish long-non-coding RNA 5
Biological Research Centre, Institute of Biochemistry, Szeged, Hungary
genes and yet another six snoRNA sequences could be found in the
ncRNA sets of closely related species. The propensity for off-target activity of the Streptococcus pyogenes
Finally, we also present our results that show the dynamic expres- Cas9 (SpCas9) has been considerably lowered by increased fidelity
sion of some snoRNAs during the early stages of zebrafish devel- (high-fidelity) variants, such as e-1, -HF12, Hypa-3, evo-4, Sniper-5,
opment and tissue-specific expression patterns for others in adults. HeF-6 and Blackjack variants7. Although exhibiting decreased
off-target propensity, they still cleave many target sequences with
T18. off-targets, which are difficult to predict and have been shown to
contain mismatches at even up to 6 positions. Furthermore, they
Multi-modal Volumetric Image Registration by Establishing
frequently show decreased on-target activities at several target
Cell Correspondence
sites that are otherwise cleaved by the wild-type SpCas9.
Manan Lalit 1,2, Mette Handberg-Thorsager 1,2, Florian Jug 3,
A systematic comparison of 20 increased fidelity nucleases revealed
Pavel Tomancak 1,2,4
that they can be ranked according to their increasing fidelity and
1
Max Planck Institute of Molecular Cell Biology and Genetics, Dresden,
their concomitant decreasing average cleavage efficiency. More
Germany; 2 Center for Systems Biology, Dresden, Germany; 3 Fondatione
instrestingly, we also found that target sequences can be ranked
Human Technopole, Milano, Italy; 4 IT4Innovations, VŠB – Technical Uni-
according to their cleavability by these nuclease variants. At one
versity of Ostrava, Ostrava-Poruba, Czech Republic
end of the target ranking there are easy-to-cleave sequences that
Keywords: Platynereis dumerilli, multi-modal registration, instance all variants can cleave, but only the highest fidelity variant cleaves
segmentation, tracking them with minimal off-target. At the other end are the hard-to-
cleave sequences that are cleaved efficiently (and without off-tar-
Early development of an animal from an egg involves a rapid
get) only by the ‘less’ increased fidelity variants.
increase in cell number and several cell fate specification events
This tendency of targets makes efforts to develop a “general” nucle-
accompanied by dynamic morphogenetic changes. In order to cor-
ase variant that would cleave every target with both high efficiency
relate the morphological changes with the genetic events, one
and high specificity pointless. Building on the above knowledge, we
typically needs to monitor the living system with several imaging
show that apparently all targets can be edited without any detect-
modalities offering different spatial and temporal resolution. Live
able, genome-wide off-target by matching the right increased fidel-
imaging allows monitoring the embryo at a high temporal resolu-
ity nuclease with the rank of the target sequence. Furhermore we
tion and observing the morphological changes. On the other hand,
demonstrate a method of choosing the most suitable SpCas9 vari-
confocal images of specimens fixed and stained for the expression
ant when high precision editing is required.
of certain genes enable observing the transcription states of an 1
Slaymaker IM et al.; Science (2016); 2 Kleinstiver BP et al.; Nature
embryo at specific time points during development with high spa-
(2016); 3Chen JS, Dagdas YS, Kleinstiver BP et al.; Nature (2017);
tial resolution. The two imaging modalities cannot, by definition, be 4
Casini A et al.; Nature Biotechnology (2018); 5Lee et al. Nature
applied to the same specimen and thus, separately obtained images
Communications (2018); 6 Kulcsár PI et al.; Genome Biology (2017);
ofdifferent specimens need to be registered. 7
Kulcsár PI et al.; Nature Communications (2020)ABSTRACTS | ORAL PRESENTATIONS 9
T20. KEYNOTE TALK 5 T22.
Whole-body connectomics and the study of motor control in The paradise fish (Macropodus opercularis) as a complemen-
larval Platynereis tary model species of behavioral and developmental genetics
Gáspár Jékely Erika Fodor 1, Tony Patelunas 2, Nóra Szabó 1, Zoltán K. Varga
Living Systems Institute, University of Exeter, UK 3
, Kata Szabó 1, Dávid Czimer 1, Anita Rácz 1, Ádám Miklósi 3,
Shawn Burgess 2 and Máté Varga 1,*
We are interested in how whole-body neural circuits coordinate 1
Department of Genetics, ELTE Eötvös Loránd University, Budapest, Hun-
the movement of an animal body. To understand this, we study
gary; 2 Translational and Functional Genomics Branch, National Human
the experimentally accessible and small larval stages of the marine
Genome Research Institute, Bethesda, MD, USA; 3 Laboratory of Transla-
annelid Platynereis. The nereid has recently emerged as a powerful
tional Behavioral Neuroscience, Institute of Experimental Medicine, Eötvös
new experimental system for neural circuits and development. With
Loránd Research Network , Budapest Hungary; 4 Department of Ethology,
serial electron microscopy, we have reconstructed the entire ner-
ELTE Eötvös Loránd University, Budapest, Hungary
vous and effector systems of the larva. Through activity imaging,
behavioural experiments, transgenes and CRISPR manipulations we In the last half century a small number of model species – each
can link genes to neurons and behaviours. In the talk I will discuss representative to a bigger taxa – have been dominating the field
the coordination of locomotor cilia, the startle circuit and the pos- of biological research. In regards to fish, zebrafish (Danio rerio) has
tural control system of the larva. The nereid is an ideal system for outshined almost every other species and while it is an extremely
connectome mapping and experiments and offers a fresh perspec- useful and convenient model organism, it has its own limitations.
tive on how the brain coordinates an entire body. The advent of new transgenic and genome editing techniques has
opened up the possibility of using nontraditional model species to
T21. address specific research hypotheses and the scientific community
Revealing secrets of Proteus anguinus by 3D X-ray imaging also realize the potential in using more unconventional experimental
Markéta Tesařová 1, Lucia Mancini 2, Edgardo Mauri 3, organisms. Broadening the field of research would allow us to have
Gregor Aljančič 4, Magdalena Năpăruş-Aljančič 4,5, Tomáš Zikmund 1, a more complex view on evolution, learn more about development
Markéta Kaucká 6, Federica Papi 3, Igor Adameyko 7, Jozef Kaiser 1 and behavior and to look for better alternatives if a question cannot
1
Central European Institute of Technology, Brno University of Technology, be answered with traditional models.
Czech Republic; 2 Elettra-Sincrotrone Trieste S.C.p.A., Basovizza, Trieste, Our aim is to develop a previously popular behavioral genetics
Italy; 3 Speleovivarium Erwin Pichl - Adriatic Speleology Society, Trieste, organism, the paradise fish (Macropodus operularis) into a new
Italy; 4 Tular Cave Laboratory, Society for Cave Biology, Kranj, Slovenia; complementary model species of developmental and behavioral
5
Research Centre of the Slovenian Academy of Sciences and Arts: Karst genetics. Recently we have established breeding protocols and
Research Institute, Postojna, Slovenia; 6 Max Planck Institute for Evolution- adapted molecular methods to work with paradise fish, and assem-
ary Biology, Plon, Germany; 7 Medical University of Vienna, Vienna, Austria bled and annotated the draft genome of this obligate air-breather
species. We present a brief summary of our results together with
Keywords: Proteus, salamander, microCT, microtomography, 3D imaging
some intriguing behavioral observations that underscore the poten-
The darkness of caves harbour an astonishing biodiversity. Cave tial of paradise fish for behavioral research.
animals show diverse morphological and physiological adaptations
which enabled them to survive in lightless and low food environ- T23.
ment. In our work, we look at the endangered Proteus anguinus Blastomeres Derived from the Vegetal Pole are the Extraem-
- the only European amphibian that lives exclusively in caves. Pro- bryonic Nutrition in Sturgeon Embryo: Transition from Holo-
teus has been under the attention of scientists for centuries. It is blastic to Meroblastic Cleavage
a top predator in the subterranean food chain, hence an excellent Mujahid Ali Shah 1, Viktoriia Iegorova 2, Roman Franěk 1,
indicator of groundwater pollution. The use of contrast-enhanced Martin Pšenička 1, Taiju Saito 1,3
X-ray computed microtomographic imaging allowed the 3D seg- 1
University of South Bohemia in Ceske Budejovice, Faculty of Fisheries
mentation of soft tissues in the head of both larval and adult spec- and Protection of Waters, South Bohemian Research Center of Aquacul-
imens to help understanding how an amphibian adapted to harsh ture and Biodiversity of Hydrocenoses, Z atisi 728/II, 389 25 Vodnany,
conditions of caves. The synergy of rare material and life sciences Czech Republic; 2 Laboratory of Gene Expression, Institute of Biotechnol-
has impacted research by proposing new tools for investigation. ogy of the Czech Academy of Sciences, Vestec, Czech Republic;
Multi-disciplinary team composed of clinicians, biologists, engineers 3
South Ehime Fisheries Research Center, Ehime University, Ainan, Ehime,
and imaging experts are currently pushing forward the understand- 798-4206, Japan.
ing of biological questions using 3D approaches. We show cartilage
of chondrocranium, the position and shape of the brain, remnant Keywords: Evolution, Fate-mapping, Vertebrates, Vegetal blasto-
eyes and olfactory epithelium with possible analysis that can be mere, Yolk cells, Extraembryonic
conducted on such 3D models. Above that, we compare Proteus Generally, amphibian (holoblastic) embryos in which all blastomeres
anguinus with Ambystoma mexicanum as an exemplary comparison contribute to one of the germ layers are preserved as a stem lineage
between cave- and surface-dwelling salamanders. Proteus angui- of vertebrates, and meroblastic cleavage has evolved independently
nus has shrouded its ecology, evolution and physiology in mysteries. in each vertebrate lineage. The increasing egg size / yolk volume is
Our high-resolution images and accurate segmentation processing the key factor for transition from holoblastic to meroblastic cleav-
could help to reveal some of these secrets and further the research age pattern. Sturgeon (Acipenser) eggs are two times larger than
on very accurate 3D microtomography data.10 ABSTRACTS | ORAL PRESENTATIONS
Xenopus laevis (amphibian); despite varying size, sturgeon embryos T25.
retain nearly same developmental characteristics as Xenopus lae- Insight into the maternal gene complement of Priapulus cau-
vis. Interestingly, due to increased yolk volume, sturgeon’s vegetal datus embryos
blastomeres are bigger and divide slower than animal blastomeres– Ferenc Tibor Kagan, Andreas Hejnol
mimic the meroblastic cleavage pattern. Therefore, detailed under- University of Bergen
standing of sturgeon cleavage pattern might reveal whether the
transition of cleavage pattern (holoblastic-meroblastic) in the acti- Within Metazoa the early development of different species is partly
nopterygian lineage is correlated with egg size / yolk volume. In orchestrated by maternally provided factors. The impact of their
this study, we found that sturgeon vegetal blastomeres formed only regulation can be highly variable (maternal factors set up body axes
primordial germ cells, and the rest of the descendants were larger in several invertebrates, but not in mammals). Their crucial role in
vegetal macromeres „yolk cells (YCs).” Morphological and pheno- development has been demonstrated in several model systems.
typic characteristics revealed that after the 1 k-cell / mid-blastula Ecdysozoan models such as Drosophila melanogaster and Caenor-
transition, YCs became transcriptionally inactive and served only habditis elegans have been rich sources of information, but their
for nutrition as larvae developed. Furthermore, inhibition of vegetal development is highly derived and therefore make it difficult to build
blastomeres showed embryogenesis without producing YCs instead hypotheses of the evolutionary ancient maternal gene repertoire.
formed acellular yolk mass similar to teleosts, which could suggest Priapulids are marine worms forming likely the sister group to
that meroblastic cleavage in the actinopterygians, teleosts, might all remaining Ecdysozoa. Their fossil records indicate their abun-
evolved by the fusion of vegetal blastomeres. dant presence in the Early Cambrian and also show a conserved
body plan. Combined with a slow nuclear and mitochondrial gene
T24. sequence evolution indicate that some priapulids represent a slowly
evolving branch.
Allele distribution modelling predicts climate change adapta-
Using RNA sequencing we sampled the early development of Pri-
tion in the bank vole
apulus caudatus. This dataset has led to an insight into the maternal
Marco Alejandro Escalante 1, Silvia Marková 1, Jeremy Searle 2 and
factors in this species. We were able to identify maternal genes that
Petr Kotlík 1
are also required for Drosophila melanogaster and Caenorhabditis
1
Laboratory of Molecular Ecology, Institute of Animal Physiology
elegans development. Furthermore, from the PAR system used by
and Genetics of the Czech Academy of Sciences, Rumburská 89, 277 21
the nematode to establish anterior-posterior axis we found only the
Liběchov, Czech Republic.; 2 Department of Ecology and Evolutionary
par3-par6-cdc42-apkc axis and not its antagonist axis par1-par2-
Biology, Cornell University, Ithaca, NY 14853, USA.
lgl1. Our analysis of the maternal gene complement shows that the
Evolutionary adaptation is an important mechanism of species’ overall conservative evolution in priapulids, is also reflected on the
response to climate variation whereby populations develop physio- level of maternal genes during the development.
logical alterations to adjust to new conditions. Therefore, evolution-
ary adaptation will likely be important for the ability of organisms T26.
to cope with future climate change. This is especially relevant to the X-ray Micro computed tomography imaging based collection of
species occurring on islands, with limited opportunities to respond reptile skull models for morphological analysis of tooth attach-
to changing environmental conditions by changing their distribu- ment
tions. Bank voles are widespread in Britain and carry two func- Michaela Kavková 1, Marie Šulcová 2, Tomáš Zikmund 1, Martin
tionally distinct haemoglobin variants HbS and HbF with a striking Pyszko 4, Jozef Kaiser 1 and Marcela Buchtová 3
north-south geographic pattern thought to be driven by environ- 1
Central European Institute of Technology, Brno University of Technol-
mental selection. The HbF variant confers increased erythrocyte ogy, Brno, Czech Republic; 2 Department of Experimental Biology, Faculty
resistance to a free-radical attack compared to HbS and is thus of Science, Masaryk University, Brno, Czech Republic; 3 L aboratory of
likely adaptive under environmental conditions favouring oxida- Molecular Morphogenesis, Institute of Animal Physiology and Genetics,
tive stress. This is important, for example, because exposure dur- v.v.i., Czech Academy of Sciences, Brno, Czech Republic; 4 Department of
ing early development to environmentally induced oxidative stress Anatomy, Faculty of Veterinary Medicine, University of Veterinary Sci-
can constrain the offspring phenotype. We therefore modelled the ences Brno, Brno, Czech Republic
current distributions of the alleles encoding HbS and HbF based
on their response to environmental factors and forecasted their Keywords: microCT, 3D imaging, reptiles, tooth-bone attachment
distributions under different future climate change scenarios. The The superorder of Lepidosaurian reptiles presents a large hetero-
areas with the currently most favourable environmental conditions genity in the teeth morphology. The main variability is based on
for HbS were predicted to be in northern Britain, while for HbF in the different tooth shape and on the type of tooth attachment
the south of the island, in agreement with their present ranges. to the underlaying bone. There are at least four different types of
However, under the future climate warming scenarios, it was fore- tooth-bone attachment ranging from mammalian-like thecodont
casted that HbF will replace HbS throughout northern Britain, which attachment found in crocodilian to the simple acrodont implanta-
indicates the species’ capacity for evolutionary adaptation to future tion occurring in some lizards. The type of tooth attachment to the
global warming. jawbone is also associated with the number of tooth generations,
which is another interesting feature varying in reptilian species.
Here, we introduce a collection of micro CT based scans of skulls of
various reptilian species. We selected model species, which possessABSTRACTS | ORAL PRESENTATIONS 11
unique, dental features among reptiles with aim to evaluate differ- MMP inhibitors. We revealed a negative effect of MMPs inhibition
ent types of tooth to bone attachment across the lepidosaurian on formation of actin ring and laminin layer resulting in wound clo-
reptiles. We introduce the analysis of tooth-bone attachment in 3D sure defects. In addition, we performed detailed expression ana-
view, which brings the new insights into evaluation of this area. The lyzes of mmps and healing markers at the whole embryo and single
obtained collection of data will be also valuable for the research of cell levels.
skull morphology in lepidosaurians and it will be suitable for the 3D
print and follow up use of the 3D printed models in education. T29.
This work was supported by CzechNanoLab Research Infrastruc- hiPSC derived cardiomyocytes: struggles and hopes
ture supported by MEYS CR (LM2018110) and Grant Agency of the Radaszkiewicz Katarzyna, Streďanská Katarína, Kohoutková Eliška,
Czech Republic grant 21-05146S. Pacherník Jiří
Department of Experimental Biology, Faculty of Science, Masaryk Univer-
T27. KEYNOTE TALK 6 sity, Brno, Czech Republic
Newt regeneration – evolution and regulation
Keywords: hiPSC, cardiomyocytes, maturation, regeneration, car-
András Simon
diovascular disease
Karolinska Institutet, Department of Cell and Molecular Biology, Stock-
holm, Sweden Cardiovascular diseases, specifically ischemic disease caused by
coronary obstruction, are the leading cause of mortality and mor-
The ability to regenerate lost body structures is present in diverse
bidity worldwide. The inadequate blood supply to heart leads to the
animal species ranging from simple organisms, such as the hydra, to
loss of cardiomyocytes and subsequently to severe structural and
complex vertebrates, such as salamanders. We aim to understand
functional defects in the myocardium, which in turn, induces heart
how animals with outstanding regenerative capabilities sense what
failure. Current therapies usually can slow down the progression of
and how much is missing in relation to the normal homeostatic state,
the disease, however standard approaches do not solve the major
and how they translate that information to the appropriate regen-
issue: depletion of cardiomyocytes and its replacement by a fibrotic
erative responses. We primarily study an aquatic salamander, the
scar. Cardiac regeneration requires new cardiomyocytes, but heart
newt, which possesses exceptional regenerative capacities among
has an extremely low regenerative ability. Therefore, cardiomyo-
adult vertebrates. Recent technical advance from our lab, such as
cytes derived from human induced pluripotent stem cells (hiPSC)
genome sequencing and in vivo editing, allow now to understand
have emerged as a potential source of therapeutic cells as well as
newt regeneration at the molecular level. In my presentation I will
a platform for disease modelling and drug discovery. Despite the
focus on mechanisms of limb regeneration with particular focus on
great promise of hiPSC-derived cardiomyocytes in medicine, many
the role of skeletal muscle.
challenges remain to be solved. Current methods of differentiation
usually generate a heterogenous mix of cardiac subtypes and non-
T28.
cardiac cells. Moreover, in vitro derived cardiomyocytes have an
The role of matrix metalloproteinases during embryonic wound immature phenotype corresponding to foetal cardiomyocytes with
healing dramatically different structural and functional properties. Due to
Paulina Kikinderova, Ravindra Naraine, Pavel Abaffy, Mikael Kubista, these major differences in hiPSC-derived and native cardiomyo-
Radek Sindelka cytes, the development of novel maturation protocols remains a
Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech major research goal. Here we show how different approaches such
Republic as long-term culture, metabolic restriction, or 3D culture affect hiP-
Keywords: wound healing, gene expression, Xenopus, matrix SC-derived cardiomyocyte phenotype.
metalloproteinases
T30.
Two types of cutaneous wound healing could be distinguished:
Sertoli cell progenitors as a promising tool for muscle regen-
embryonic and adult. Embryonic healing in contrast to adult is
eration
faster and scar-less and consists of early, middle, and late phases.
Vladimír Krylov 1, Aneta Wróblová 2, Jakub Onhajzer 3,
Actin ring is formed during the early phase and de novo expression
Tereza Tlapáková 1
of healing specific genes is initiated also during middle phase. Our 1
Charles University, Faculty of Science, Department of Cell Biology,
understanding about late phase remains elusive. Essential role in
Viničná 7, Prague 2, Czech Republic; 2 Clinic for Reproductive Medicine
the wound healing is given to matrix metalloproteinases (MMPs).
IVF Cube, Evropská 423, Prague 6, Czech Republic; 3 Institute of Molec-
These remodeling enzymes are important for releasing the cyto-
ular Genetics of the Czech Academy of Sciences, L aboratory of Cell and
kines, inflammatory cells, apoptosis, and degradation of extracel-
Developmental Biology, Vídeňská 1083, Prague 4, Czech Republic
lular matrix. Our laboratory performed temporal RNA sequencing
of the healing tissue using tailbud and swimming tadpole embryos. Keywords: XtiSCs, Xenopus tropicalis, muscle regeneration
Results showed predominant expression of four mmps: 1, 7, 8 and 9.
Muscle regeneration is a limited process in higher vertebrates
Injury or amputation caused the upregulation and their expression
including human. Striated musculature is renewed from satellite
level peaked at 3-6 hours post injury, which corresponds to late
Pax7+ cells localized beneath the basal lamina surrounding individ-
phase of healing. We used in situ hybridization and scRNA-Seq to
ual myofibers. Heart healing after ischemia or infarct is restricted
visualize cells expressing mmps. We also studied functional relation-
to formation of fibrotic scar without substantial proliferation of car-
ships between each selected mmps by using specific and general
diomyocytes. We succeeded in the establishment of culture of Ser-You can also read
