Zurich Center for Integrative Human Physiology

ZIHPNewsAnimal experiments are one of the key requisites for progress in medical research. Rodent models enable bet-ter insight into complex biological processes. Newly developed tech-niques and high-throughput pheno-typing procedures – like imaging modalities, miniaturized implantable devices and laboratory analyses – enable investigations of many pro-cesses in living animals.

All of these techniques have in com-mon that the acquisition of the equipment is costly, operation and servicing is technically demanding, and downscaling to small rodents is most often challenging. Skilled staff with appropriate know-how is nee-ded for their optimal utilization. As a consequence, the individual resear-cher most often does not have the ability to acquire all necessary equip-ment, cover its running costs and pay technical staff for operation.

→ Zurich Integrative Rodent Physio-logy (ZIRP) provides infrastructure and equipment with a focus on ana-lyses of rodent physiology on a shared resources basis, thus making complex techniques available at low cost to all researchers.

ZIRP’s mission: facilitate and improve ex-perimental workZIRP pools the researchers’ needs and facilitates their work by organizing and maintaining key infrastructure, providing training and support, and offering a number of basic services.

ZIRP’s services are organized to faci-litate customized solutions. In addi-tion to special technical platforms, a number of basic services are provi-ded. For example, • workspace, completely equipped

«ready to use» or free space to build up own equipment

• sample collection• administration of substances and• surgical services.

ZIRP’s strategy: flexible, research-driven approach - individually tailored solutionsZIRP offers flexible solutions and in-dividual strategies in close coopera-

tion with researchers by adapting procedures or combining different techniques with respect to indivi-dual requirements. ZIRP’s skilled staff utilizes a broad array of stan-dard experimental procedures, ad-apted to the characteristics of small rodent models.

ZIRP’s vision: one-stop - all services from one sourceZIRP seeks to offer all services from one common source with all experi-mental work performed within the ZIRP - from transferring the ani-mals into ZIRP’s animal housing facility and performing different experiments to having the data available ready for analysis. ZIRP continuously upgrades its services and the ZIRP staff members are per-manently refining existing tech-niques – always considering the re-searchers’ needs.

Zurich Integrative Rodent Physiology (ZIRP): Bundled competences for research

5-2014 • July 2014

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ZIHPNews 05-2014

Imaging Platform

Non-invasive imaging modalities offer numerous options for morpho-logic analyses and tracking of biolo-gical processes like disease progres-sion or metabolic pathways. The → ZIRP imaging platform provides a number of state-of-the-art imaging devices as well as associated servi-ces and support.

Micro CTIn vivo micro-computed tomogra-phy is a non-invasive tool to visua-lize the animal’s morphology. Com-mon applications include imaging of the skeleton or bone samples, lung, cardiovascular system, soft tissue and tumors.

For soft tissue imaging a multitude of contrast agents are available, e.g. for the depiction of vessels or organs (Figure 1).

High throughput imaging at low ra-diation doses is ideally suited for morphological phenotyping (Figure 2). High resolution scans are the me-thod of choice for quantitative mor-phometric image analyses (Figure 1).

Optical ImagingBioluminescence and fluorescence imaging allow the visualization and quantification of biological processes in the living animal in real-time.

Bioluminescence imaging utilizes native light emission from biolumi-nescing organisms. The DNA enco-ding the luminescent protein (for example firefly luciferase) is incor-porated into the laboratory animal. For light emission the corresponding substrate (for example D-luciferin) needs to be injected into the animal prior to imaging (Figure 3, left).

Bioluminescence imaging is com-monly used for in vivo studies of cancer progression, development or cell migration, or infection.

In addition, bioluminescence tomo-graphy allows the quantitative ana-lysis of volumes and therefore, the precise three-dimensional follow up of disease progression. This is useful for the growth and spread of a tumor (Figure 3, left).

Fluorescence imaging requires a spe-cific fluorescing agent able to absorb and emit light of a certain wave-length which needs to be injected

Figure 2: Skull phenotyping in mice: normal (left) versus malformed skull (right).

Figure 1: Visualization of kidney cysts in a rat using contrast agent, kidneys are cut longitudinally to better visualize the cysts.

Figure 3: Bioluminescence imaging of a mouse lung tumor, co-registration of 3D micro CT and bioluminescence data (left) and visualization of subcutaneous tumor vascularization with fluorescence imaging using a targeted fluorescent agent (An-gioSense 680, Perkin Elmer, right).

«Optical imaging of tumour cells is an invaluable tool for long term

quantitative monitoring of cancer growth and spread in living mice.»

Prof. Ian Frew, ZIHP Assistant Professor Institute of Physiology, University of Zurich

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ZIHPNews 05-2014

Telemetry Platform

Radiotelemetric transmitters enable the continuous and contact-free col-lection of physiological data in con-scious and freely moving animals. For example respiration rate, body temperature, activity, pressure or biopotential signals can be monito-red (Figure 4). The implanted teleme-try device transmits the digitalized data via radio frequency signals to a nearby receiver plate.

The advantages are multifaceted: Since the miniaturized transmitter and all catheters and leads are com-pletely internalized, the risk of wound infection is eliminated, animals can be group housed in their home cage di-

rectly after implantation and hand-ling of animals is minimized in order to avoid stress related artefacts.

Telemetric measurements can be combined with other techniques like running wheels, climate chambers etc. Furthermore, instrumented ani-mals can serve as their own controls.

The → ZIRP telemetry platform offers four different transmitter types for the acquisition of pressure and bio-potential signals, body temperature and activity in mice and rats as well as the relevant hard- and software system for data acquisition and ana-lysis. Other transmitters can be im-planted on request. ZIRP’s entire service includes transmitter refur-bishment, surgical implantation and postoperative care with the animal handed over «ready to use».

«ZIRP’s telemetry platform allows us to match in vitro organ chamber experiments with in vivo blood pressure measurement.»

Dr. Elvira Haas, Research Unit, Div. of Internal Medicine, University Hospital Zurich

«Telemetry is a key technology in hypertension research. ZIRP provi-des not only the equipment but also the skills and the know-how for successful telemetry.»

Dr. Branko Simic Center for Molecular Cardiology,

University Hospital Zurich

prior to imaging. Targeted fluores-cent markers actively bind to target structures, thus allowing the selec-tive visualization of specific tissues or processes like tumor vasculariza-tion or inflammatory processes (Fi-gure 3, right).

The ZIRP imaging platform is equip-ped with the following instruments:• IVIS Spectrum optical imaging

system (bioluminescence and flu-orescence, Perkin Elmer)

• Quantum Fx micro CT (Perkin El-mer)

• Body composition analyzer (EchoMRI) for the measurement of whole body fat, lean, free water, and total water masses in living awake rodents.

Additionally, ZIRP offers solutions for contrast agent application, e.g. catheter placement and perfusion pumps for continuous intravenous infusion.

Corresponding image analysis soft-ware and server space for data ex-change and storage is offered in co-operation with the Center for Microscopy and Image Analysis (ZMB) of the University of Zurich.

Figure 4: Course of body temperature, motor acti-vity and blood pressure in a rat during 30 days after transmitter implantation.

«MicroCT imaging gave us a wealth of phenotypic information about our mice - including new aspects that we didn’t even realise.»

Prof. Steven A. Brown Institute of Pharmacology and Toxicology,

University of Zurich

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ZIHPNews 05-2014

Laboratory Platform

Biochemical analyses of biological fluids offer insight into the function of various organ systems. Animal models can be assessed by analy-zing a multitude of different para-meters in one sample.

In rodents such an evaluation is often hampered by the small volume of their body fluids – a limitation which is even more critical in case of repea-ted measurements during longitudi-nal studies or the analysis of a large number of parameters in one sample.

High quality analyses of small samplesThe → ZIRP laboratory platform of-fers high quality analyses for a wide range of biochemical and toxicologi-cal parameters in small volumes of any biological fluid. All analyses are tailored to the researcher’s specific needs. Consistency, high accuracy and technical attention to precious samples are the essential priorities.

All methods are ideally suited for the analysis in rodent models. Additio-nally, the laboratory platform has the capacity for high-throughput analy-sis of human samples from large po-pulation cohorts where small sample size could be also a limitation (e.g. genotype-phenotype correlations).

The platform utilizes liquid che-mistry technologies, which allow the acquisition of a large array of reliable biochemical analyses in minimal vo-lumes (usually < 10-15 µl) with a high throughput (> 200 analyses per hour) in combination with an automated calibration system.

The platform provides the following equipment:• biochemical analyser (Beckman

UniCel DxC 800 Synchron) • osmometers (Advanced Osmome-

ter 2020)• Mettler-Toledo T50 microtitrator• Radiometer ABL 80 CO-OX blood

gas analyzer The platform also offers a series of special protocols such as those for quantification of bicarbonate, uro-modulin, cyclic AMP, vasopressin, ammonium as well as the possibility to develop new assays.

Challenging sample collectionSample collection can be challenging in small rodents. Pre-analytical er-rors occurring during sample collec-tion or post-processing are a major source of missed or wrong diagno-ses. Therefore, ZIRP not only offers help with any type of sample collec-tion but also a collection of tubes sui-ted for optimal processing of small amounts of different types of biolo-gical fluids.

Figure 5: Automated analysis of serum samples

Zurich Integrative Rodent Physiology (ZIRP)Zurich Integrative Rodent Physiology (ZIRP) is an interdisciplinary core facility at the Irchel campus of the University of Zurich. Its mission is to support and strengthen research activities in the field of integrative physiology by providing infrastructure and know-how for the advanced analysis of rodent physiology. ZIRP is open to all researchers from the University of Zurich, the ETH, other research institutions and companies. ZIRP is currently supported by the ZIHP, the NCCR Kid-ney.CH and the Institute of Physiology, UZH.

General contact→ www.zirp.uzh.ch 044 635 50 95Dr. P. Seebeck → petra.seebeck@uzh.chWinterthurerstrasse 190, 8057 Zürich

Imaging and Telemetry PlatformS. Pfundstein → svende.pfundstein@uzh.chLaboratory PlatformN. Nägele → nadine.naegele@uzh.ch

«This platform enables analyses of numerous parameters in small sample volumes with high accuracy.»

Prof. Olivier Devuyst Institute of Physiology, University of Zurich

«ZIRP's mission is to facilitate and improve experimental work in rodent models.»

Dr. Petra Seebeck , ZIRP Manager

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ZIHPNews 05-2014

Activity Report 2013The → Activity Report 2013 of the University Research Priority Program «Integrative Human Physiology» can now be downloaded from the ZIHP website.

10th ZIHP Symposium: August 29, 2014

The spacious foyer at the Häldeliweg opens its doors again for an exciting day of science where basic resear-chers and clinical scientists get to-gether to present and discuss their research in fields related to human physiology. In particular, young re-searchers will have the opportunity to present their cutting-edge results in oral and poster presentations.

A major part of the presentations is given by students of the → imMed PhD Program and originates from ZIHP funded → cooperative projects.

Cash prizes will be awarded for the best presentation and the best pos-ters. The ceremony of the ZIHP Award will take place at the sympo-sium.

Don’t miss the opportunity for an intensive scientific exchange in a re-laxed atmosphere!

→ Website and program of the symposium

Deadline for → registration is Friday, August 15, 2014.

Keynote speakers:→ Prof. Friedo Dekker, Dept. of Cli-nical Epidemiology, Leiden Univer-sity Medical Center, NetherlandsFraud, plagiarism, and other «minor» problems in science

→ Prof. Heribert Schunkert, German Heart Center, Munich, Germany A look into the genome of myocardial infarction

→ Prof. Bengt Kayser, Institute of Sports Sciences, University of Lau-sanne, SwitzerlandStill the better polypill: exercise

Recent publicationsBaruffol C, Jordi J, Camargo S, Radovic T, Herzog B, Fried M, Schwizer W, Verrey F, Lutz TA, Steingoetter A: → L-lysine dose dependently delays gastric emptying and increases intestinal fluid volume in humans and rats. [Epub ahead of print], 2014

Daryadel A, Bogdanova A, Gassmann M, Mueller X, Zünd G, Seifert B, Lehalle C, Frossard N, Tavakoli R: → Multikinase in-hibitor sorafenib prevents pressure over-load-induced left ventricular hypertro-phy in rats by blocking the c-Raf/ERK1/2 signaling pathway. J Cardiothorac Surg 9(1): 81, 2014

Myakala K, Motta S, Murer H, Wagner CA, Koesters R, Biber J, Hernando N: → Renal-specific and inducible depletion of NaPi-IIc/Slc34a3, the cotransporter mutated in HHRH, does not affect phos-phate or calcium homeostasis in mice. Am J Physiol Renal Physiol 306(8): F833-43, 2014

Wueest S, Item F, Boyle CN, Jirkof P, Cesa-rovic N, Ellingsgaard H, Böni-Schnetzler M, Timper K, Arras M, Donath MY, Lutz TA, Schoenle EJ, Konrad D: → Interleu-kin-6 contributes to early fasting-indu-ced free fatty acid mobilization in mice. Am J Physiol Regul Integr Comp Physiol 306(11): R861-7, 2014

EventsJuly 8, 2014 - PhD Thesis Defense → The role of white adipose tissue in the development of obesity-associated insu-lin resistance and endogenous fat mass control Michael Wiedemann, University Children's Hospital Zurich

July 11, 2014 - Wagi Seminars → Mechanisms and modulation of he-moglobin toxicity Prof. Dominik Schaer, Lab of Internal Medicine, USZ

21.-22. August 2014 → Rheuma-Top 2014

August 29, 2014 → 10th Symposium of the ZIHP

1.-5. September 2014 → Medidays Zürich

imMed PhD Program graduate courseJuly 10/11, 2014 → Mouse physiology and patho-physiology Please register asap by e-mail to preisig@zihp.uzh.ch.

Congratulations!ZIHP-Mitglied Beatrice Beck Schimmer erhielt von der Deutschen Gesellschaft für Anästhesiologie und Intensivmedizin (DGAI) die → Manfred-Specker-Medaille verlie-hen für ihre herausragenden Ver-dienste um den wissenschaftlichen Nachwuchs der Anästhesiologie als Mentorin im Programm der DGAI.

ZIHP-Mitglied Raghvendra Dubey wurde auf den 1. August 2014 zum → ausserordentlichen Professor ad personam für Molekulare Reproduk-tionsendokrinologie an der Medi- zinischen Fakultät der Universität Zürich ernannt.

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ZIHPNews 05-2014

ZIHP cooperative sprint projects starting by July 2014

Four new ZIHP cooperative sprint projects start by July 2014. At least two ZIHP full or junior members joined forces and defined projects with a balanced representation of basic and clinical research. Funding of these projects is limited to one position for a PhD or a MD-PhD student and consumables.

→ Translational studies of endocrine and brain controls of eating in rats and pre and postmenopausal womenObesity is a world-wide epidemic associated with severe co-morbidi-ties such as cardiovascular disease and type 2 diabetes. Women are more vulnerable than men to deve-lop obesity, and among women, the postmenopausal population is at even higher risk. In working towards a cure for obesity in women, it is im-perative to know (a) how eating is controlled in premenopausal women and (b) based on this physiology, what is amiss in postmenopausal women. Translational studies will be performed to investigate the peri-pheral and brain controls of eating in pre- and postmenopausal women.

→ Erythropoietin regulates erythro-ferron regulates hepcidin regulates iron uptake: testing this hypothesis in man and mouseHypoxia induces erythropoiesis, a process requiring high iron levels. Upon hypoxia, erythropoietin (Epo) is secreted and reaches the bone mar-row where it binds to its receptor present on erythroid precursors. On the other hand, the hepatic hormone hepcidin regulates iron homeostasis. Very recently, a novel erythroid fac-tor termed erythroferron (ErFe) was presented. Evidence was provided that Epo induces ErFe expression in erythroblasts that ultimately inhibi-ted hepcidin production in hepato-cytes, thereby enabling iron uptake in mice. In already available blood samples it can now be investigated

as whether Epo controls ErFe (i) in volunteers acutely exposed to 4559m of altitude, (ii) in volunteers that re-ceived a single high Epo dosis and (iii) in constitutively Epo-overex-pressing mice.

→ Role of islet amyloid polypeptide (IAPP) derived deposits in vascular dysfunction in diabetesThe deposition of amyloid in specific tissues is a feature of diseases like type 2 diabetes (T2DM; in the pan-creas) and Alzheimer disease (in the brain). The presence of amyloid de-posits has cytotoxic consequences and seems to be the unifying patho-

physiological mechanism in very different conditions such as TDM2 or Alzheimer’s. Amyloid in T2DM is derived from islet amyloid polypep-tide (IAPP) and has recently been found in the wall of blood vessels. In this project, the potential functional implications of these deposits in the vessel wall for endothelial dysfunc-tion in T2DM will be studied using rodent models that develop human-IAPP derived amyloid.

→ A protective role of SIRT6 in arte-rial thrombosis? – From mice to menArterial thrombosis may lead to myocardial infarction, a major cause of mortality in Western countries. Endothelial pro-inflammatory pro-cesses involving NF-κB are driving forces of thrombogenesis. Sirtuin 6 (Sirt6) is a nuclear histone deacety-lase with beneficial effects on inflam-mation. Its role in thrombosis and its protagonist cells are unknown. It is hypothesized that Sirt6 diminishes arterial thrombosis by decreasing in-flammatory NF-κB and c-Jun-medi-ated signaling in endothelial cells. This hypothesis will be tested by ful-filling translational aims at the cellu-lar and organismal level using Sirt6 loss-of-function models, as well as samples from patients with acute co-ronary syndromes (ACS).

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ZIHPNews 05-2014

New open PhD positions→ Molecular mechanisms and inter-actions in methylmalonic aciduria Division of Metabolism, Children’s Research Center, University Children's Hospital Zurich

→ In vitro fabrication and bio-func-tional characterization of a native-analogous endothelialized bio-multi-layer superimposed onto synthetic hyperelastic PDMS-memb-ranes Swiss Center for Regenerative Medicine, USZ

→ The role of hypoxia inducible fac-tors at the blood brain barrier Institute of Veterinary Physiology, UZH

→ The role for innate lymphoid cells in the pathogenesis of Crohn’s di-sease associated fistulae Division of Gastroenterology and Hepatology, USZ

→ Pancreatic regeneration Division of Visceral and Transplant Surgery Research, USZ

→ The interplay between angioten-sin II, Wnt signalling and TGF-beta in myocardial fibrogenesis and heart dysfunction in systemic sclerosis Research of Systemic Autoimmune Diseases, Divison of Rheumatology, USZ

→ Molecular regulation of stem cell-mediated bone regeneration Center for Applied Biotechnology and Molecular Medicine, UZH

→ Bone regeneration by titanium/magnesium based scaffolds Division of Cranio-Maxillofacial and Oral Surgery/Bioengineering, USZ

Press review

→ «Die langfristige Entwicklung ei-nes Kindes kann man nicht voraus-sagen»An der Universität Zürich Irchel tref-fen sich etwa 400 Fachleute zur tra-ditionellen Tagung der Abteilung Entwicklungspädiatrie des Kinder-spitals Zürich. Oskar Jenni, ZIHP-Mitglied und Leiter der Abteilung, warnt vor vorschnellen Diagnosen und überhöhten gesellschaftlichen Erwartungen an das Kind. UZH News, 24. Juni 2014

→ Den Tumor durchlöchertEin Schweizer Ärzteteam - unter ih-nen ZIHP-Mitglied Ernst Martin - hat erstmals einen Patienten, der an ei-nem bösartigen Hirntumor leidet, mit fokussiertem Ultraschall behandelt.NZZ am Sonntag, 15. Juni, 2014

→ Wenn Lebensmittel krank machenWeizen-Unverträglichkeiten sind weit verbreitet. Eine Zöliakie zu er-kennen, erfordert mitunter erhebli-chen detektivischen Spürsinn, er-klärt der ZIHP-Forscher und Gastroenterologe Michael Fried.Neue Zürcher Zeitung, 11. Juni 2014

→ Brasilien 2014: Weltmeisterschaft im Sauna-DampfbadEs sind die klimatischen Bedingun-gen, die vielen Fussballspielern an der Weltmeisterschaft in Brasilien besonders zu schaffen machen. ZIHP-Mitglied Carsten Lundby zeigt, wie unser Organismus auf sol-che Extremsituationen reagiert.10vor10, SRF, 6. Juni 2014

→ Den Weg zur Forschung ebnenDie Medizinische Fakultät der Uni-versität Zürich hat ein neues Förde-rungsprogramm für Medizinerinnen und Mediziner initiiert. Geleitet wird es von ZIHP-Mitglied Beatrice Beck Schimmer. Unter dem Label «Filling the Gap» erhalten in diesem Jahr 24 Nachwuchsforschende – da-von 17 Frauen – die Möglichkeit, ihre Arbeit in der Klinik zu reduzieren. Zugunsten ihrer Forschung.UZH News, 4. Juni 2014

→ Mathematik hilft PsychiatrieKönnen mathematische Modelle von Gehirnvorgängen die Behandlung psychiatrischer Erkrankungen ver-bessern? Die geladenen Fachleute am ersten Zürcher Meeting zur «Com-putational Psychiatry» zeigten sich davon überzeugt. Einer der Wegbe-reiter dieser Forschungsrichtung ist ZIHP-Mitglied Klaas E. Stephan.UZH News, 28. Mai 2014

Imprint Zurich Center for Integrative Human Physiology (ZIHP), Winterthurerstr. 190, 8057 Zurich Editorial staff: M. Seebauer, P. Seebeck, C. Giger, H. Preisig, G. Roth and M. Gassmann URL: http://www.zihp.uzh.ch, E-Mail: info@zihp.uzh.ch, Tel +41 44 635 50 88/47

Other open positions→ Koordinator/in 50% (Mutter-schaftsvertretung, befristet auf 5 Monate) Zürcher Zentrum für Integrative Hu-manphysiologie (ZIHP)

→ Lab internship in metabolic medi-cine for a summer student University Children's Hospital Zurich