In Vivo Microscopy Unit (IVMU)

Research

For decades, mechanistic studies on brain circuits had to be performed in reduced preparations: acute brain slices, organotypic slice cultures, dissociated neurons and glial cells. While these in vitro and ex vivo approaches may greatly simplify the research and yield a wealth of valuable data, they may also introduce a variety of artifacts because brain cells behave very differently in the intact brain as compared to the reduced preparations. With recent advances in deep brain imaging/recording/manipulation techniques (such as two-photon microscopy, multichannel electrophysiology and optogenetic stimulation), it has finally become possible to monitor and activate individual cells, and even individual synapses, within the brain of anesthetized rodents. Furthermore, since anesthesia is known to distort the brain function, the most recent trend is to maximize the relevance of in vivo research by performing imaging and recording without anesthesia, i.e. in awake and behaving mice. In order to provide access to these advanced techniques for its research groups, the Neuroscience Center has established in 2013 a new facility - In Vivo Microscopy Unit (IVMU).
The IVM Unit has a chief purpose of enabling advanced research of NC groups by providing expertise in the methodologies of in vivo microscopy (primarily, two-photon microscopic imaging), animal surgery and data analysis. The Unit is organized as a collaborative core facility, whereby the IVMU personnel provides consultancy and training to NC personnel and closely supervises their in vivo microscopy experiments.  The Unit is coordinated by a part-time Principal Investigator (Leonard Khiroug) and is staffed with a full-time Animal Surgery Technician (Marina Tibeikina) and a part-time Postdoctoral Researcher (Evgeny Pryazhnikov). Key equipment of the IVMU consists of a two-photon microscopy setup, an animal surgery setup, rodent housing rigs (Scantainers) and supporting equipment for standard lab procedures (biochemistry and molecular biology).

People

Unit leader: Docent Leonard Khiroug, PhD, principal investigator
Animal Surgery Technician: Marina Tibeykina, PhD
Postdoctoral Researcher: Evgeny Pryazhnikov, MD, PhD

Selected Publications

Kislin M, Mugantseva E, Molotkov D, Kulesskaya N, Khirug S, Kirilkin I, Pryazhnikov E, Kolikova J, Toptunov D, Yuryev M, Giniatullin R, Voikar V, Rivera C, Rauvala H, Khiroug L. (2014) Flat-floored air-lifted platform: a new method for combining behavior with microscopy or electrophysiology on awake freely moving rodents. J Vis Exp. (88):e51869. [abstract]

Paveliev M, Kislin M, Molotkov D, Yuryev M, Rauvala H, Khiroug L. (2014) Acute brain trauma in mice followed by longitudinal two-photon imaging. J Vis Exp. (86). [abstract]

Pryazhnikov E, Kislin M, Tibeykina M, Toptunov D, Ptukha A, Shatillo A, Gröhn O, Giniatullin R, Khiroug L. (2014) Opposite reactivity of meningeal versus cortical microvessels to the nitric oxide donor glyceryl trinitrate evaluated in vivo with two-photon imaging. PLoS One. 9(2):e89699. [abstract]

Molotkov D, Zobova S, Arcas JM, and Khiroug L. (2013) Calcium-induced outgrowth of astrocytic peripheral processes requires actin binding by Profilin-1. Cell Calcium 53(5-6):338-48. [abstract]

Kremneva E, Kislin M, Kang X, Khiroug L. (2012) Motility of astrocytic mitochondria is arrested by Ca(2+)-dependent interaction between mitochondria and actin filaments. Cell Calcium doi:pii: S0143-4160(12)00174-1. [abstract]

Rapinen S., Pryazhnikov E., Khiroug L., Ericson M., Yliperttula M and Urtti A. (2012) Organotypic cell cultures and two-photon imaging: tools for in vitro and in vivo assessment of percutaneous drug delivery and skin toxicity. J Control Release 161(2):656-67. [abstract]

Yuryev M and Khiroug L. (2012) Dynamic longitudinal investigation of individual nerve endings in the skin of anesthetized mice using in vivo two-photon microscopy. J Biomed Opt 7(4):046007. [abstract]

Kolikova J, Afzalov R, Surin A, Lehesjoki A-E and Khiroug L. (2011) Deficient mitochondrial Ca2+ buffering in the Cln8mnd mouse model of neuronal ceroid lipofuscinosis. Cell Calcium 50: 491-501. [abstract]

Pryazhnikov E, Fayuk D, Niittykoski M, Giniatullin R and Khiroug L. (2011) Unusually strong temperature dependence of P2X3 receptor traffic to the plasma membrane. Front Cell Neurosci 5: 27. [abstract]

Molotkov D, Yukin A, Afzalov R and Khiroug L. (2010) Gene delivery to postnatal rat brain by non-ventricular plasmid injection and electroporation. J Vis Exp pii: 2244. doi: 10.3791/2244.[abstract]

Khirug S, Pryazhniko E, Colema, SK, Jeromin A, Keinänen K and Khiroug L. (2009) Dynamic visualization of membrane-inserted fraction of pHluorin- tagged channels using repetitive acidification technique. BMC Neurosci. 10: 141.  [abstract]
   
Pryazhnikov E and Khiroug L. (2008). Sub-micromolar increase in [Ca2+]i triggers delayed exocytosis of ATP in cultured astrocytes. Glia 56: 38-49. [abstract]

Kolikova J, Afzalov R, Giniatullina A, Surin A, Giniatullin R, and Khiroug L. (2006). Calcium-dependent trapping of mitochondria near plasma membrane in stimulated astrocytes. Brain Cell Biol. 35: 75-86. [abstract]

Khirug S, Huttu K, Ludwig A, Smirnov S, Voipio J, Rivera C, Kaila K, and Khiroug L. (2005). Distinct properties of functional KCC2 expression in immature mouse hippocampal neurons in culture and in acute slices. Eur. J. Neurosci. 2: 899-904. [abstract]