|Last Name:||Witte||Position:||Wissenschaftlicher Mitarbeiter|
|Akademischer Titel:||Dr. rer. nat.||Tel.:||+49-(0)551/39-66876|
- Seit August 2010
Postdoc an der Universitätsmedizin Göttingen der Georg-August-Universität, Zentrum Anatomie, Institut Neuroanatomie unter der Leitung von Prof. Dr. J. Staiger
- 2010 Promotion
in der Arbeitsgruppe Neurobiologie an der Universität Leipzig unter Leitung von Prof. Dr. R. Rübsamen, Thema: "Differenzierung und Reifung GABAerger und glycinerger Neurotransmission im anteroventralen Cochleariskern der Wüstenrennmaus (Meriones unguiculatus)"
- 2008 - 2009
wissenschaftl. Mitarbeiter an der Universität Leipzig in der Arbeitsgruppe Neurobiologie von Prof. Dr. R. Rübsamen
- April 2005 - April 2010
Mitglied im Internationales Promotionsprogramm (IPP) "Von der Signalverarbeitung zum Verhalten"http://www.uni-leipzig.de/zhs/ipp/
- April 2005 - April 2010
Mitglied im Graduiertenkolleg GRK 1097/1 "Interneuro" http://www.uni-leipzig.de/~ineuro/
- 2005 Diplomarbeit
in der Arbeitsgruppe Neurobiologie an der Universität Leipzig unter Leitung von Prof. Dr. R. Rübsamen, Thema:"Entwicklung inhibitorischer Einflüsse im Cochleariskern: eine Patch-clamp Studie"
- 2003 Projektarbeit
in der Arbeitsgruppe Verhaltensbiologie an der Universität Leipzig unter Leitung von PD Dr. P. A. Stevenson und Prof. Dr. K. Schildberger, Thema: "Die aminerge Modulation des Mandibel-Schließermuskels bei der Grille (Gryllus bimaculatus)"
- 2001 - 2005
Hauptstudium im Fach Biologie an der Universität Leipzig mit den Hauptfächern: Neurobiologie, Genetik, Verhaltensphysiologie, Spezielle Zoologie und Biochemie
- 1999 - 2001
Grundstudium im Fach Biologie an der Universität Leipzig
(II) Etablierung der Methode zur Identifizierung von verbundenen GABAergen Interneuronen in den ausgewählten (I) neokortikalen Schichten im akuten Hirnschnitt
(III) Simultane "gepaarte" Patch-Clamp-Ableitungen von Martinotti-Zellen und den vorgeschalteten inhibitorischen Interneuronen, sowie deren Charakterisierung nach der Petilla Terminologie
Nicotine reverses hypofrontality in animal models of addiction and schizophrenia.
Koukouli,F.; Rooy,M.; Tziotis,D.; Sailor,K.A.; O'Neill,H.C.; Levenga,J.; Witte,M.; Nilges,M.; Changeux,J.P.; Hoeffer,C.A.; Stitzel,J.A.; Gutkin,B.S.; DiGregorio,D.A.; Maskos,U..
Nature Medicine, 2017.
Intracortical Network Effects Preserve Thalamocortical Input Efficacy in a Cortex Without Layers.
Guy,J.; Sachkova,A.; Möck,M.; Witte,M.; Wagener,R.J.; Staiger,J.F..
Cerebral Cortex DOI 10.1093/cercor/bhw281, 2016.
Layer IV (LIV) of the rodent somatosensory cortex contains the somatotopic barrel field. Barrels receive much of the sensory input to the cortex through innervation by thalamocortical axons from the ventral posteromedial nucleus. In the reeler mouse, the absence of cortical layers results in the formation of mispositioned barrel-equivalent clusters of LIV fated neurons. Although functional imaging suggests that sensory input activates the cortex, little is known about the cellular and synaptic properties of identified excitatory neurons of the reeler cortex. We examined the properties of thalamic input to spiny stellate (SpS) neurons in the reeler cortex with in vitro electrophysiology, optogenetics, and subcellular channelrhodopsin-2-assisted circuit mapping (sCRACM). Our results indicate that reeler SpS neurons receive direct but weakened input from the thalamus, with a dispersed spatial distribution along the somatodendritic arbor. These results further document subtle alterations in functional connectivity concomitant of absent layering in the reeler mutant. We suggest that intracortical amplification mechanisms compensate for this weakening in order to allow reliable sensory transmission to the mutant neocortex
Parvalbumin- and vasoactive intestinal polypeptide-expressing neocortical interneurons impose differential inhibition on Martinotti cells.
Walker F, Möck M, Feyerabend M, Guy J, Wagener RJ, Schubert D, Staiger JF, Witte M.
Nature Comunications 7:13664 (DOI: 10.1038/ncomms13664, 2016.
Disinhibition of cortical excitatory cell gate information flow through and between corticalcolumns. The major contribution of Martinotti cells (MC) is providing dendritic inhibition toexcitatory neurons and therefore they are a main component of disinhibitory connections.Here we show by means of optogenetics that MC in layers II/III of the mouse primarysomatosensory cortex are inhibited by both parvalbumin (PV)- and vasoactive intestinalpolypeptide (VIP)-expressing cells. Paired recordings revealed stronger synaptic inputonto MC from PV cells than from VIP cells. Moreover, PV cell input showed frequencyindependentdepression, whereas VIP cell input facilitated at high frequencies. Thesedifferences in the properties of the two unitary connections enable disinhibition with distincttemporal features.
Thalamocortical Connections Drive Intracortical Activation of Functional Columns in the Mislaminated Reeler Somatosensory Cortex .
Robin J. Wagener, Mirko Witte, Julien Guy, Nieves Mingo-Moreno, Sebastian Kügler, Jochen F. Staiger.
Cerebral Cortex, 2015.
Neuronal wiring is key to proper neural information processing. Tactile information from the rodent's whiskers reaches the cortex via distinct anatomical pathways. The lemniscal pathway relays whisking and touch information from the ventral posteromedial thalamic nucleus to layer IV of the primary somatosensory "barrel" cortex. The disorganized neocortex of the reeler mouse is a model system that should severely compromise the ingrowth of thalamocortical axons (TCAs) into the cortex. Moreover, it could disrupt intracortical wiring. We found that neuronal intermingling within the reeler barrel cortex substantially exceeded previous descriptions, leading to the loss of layers. However, viral tracing revealed that TCAs still specifically targeted transgenically labeled spiny layer IV neurons. Slice electrophysiology and optogenetics proved that these connections represent functional synapses. In addition, we assessed intracortical activation via immediate-early-gene expression resulting from a behavioral exploration task. The cellular composition of activated neuronal ensembles suggests extensive similarities in intracolumnar information processing in the wild-type and reeler brains. We conclude that extensive ectopic positioning of neuronal partners can be compensated for by cell-autonomous mechanisms that allow for the establishment of proper connectivity. Thus, genetic neuronal fate seems to be of greater importance for correct cortical wiring than radial neuronal position.
Characterizing VIP Neurons in the Barrel Cortex of VIPcre/tdTomato Mice Reveals Layer-Specific Differences.
Prönneke A, Scheuer B, Wagener RJ, Möck M, Witte M, and Staiger JF.
Cereb. Cortex (2015) 25 (12): 4854-4868. doi: 10.1093/cercor/bhv202 , 2015.
abstract pdf link
Neocortical GABAergic interneurons have a profound impact on cortical circuitry and its information processing capacity. Distinct subgroups of inhibitory interneurons can be distinguished by molecular markers, such as parvalbumin, somatostatin, and vasoactive intestinal polypeptide (VIP). Among these, VIP-expressing interneurons sparked a substantial interest since these neurons seem to operate disinhibitory circuit motifs found in all major neocortical areas. Several of these recent studies used transgenic Vip-ires-cre mice to specifically target the population of VIP-expressing interneurons. This makes it necessary to elucidate in detail the sensitivity and specificity of Cre expression for VIP neurons in these animals. Thus, we quantitatively compared endogenous tdTomato with Vip fluorescence in situ hybridization and αVIP immunohistochemistry in the barrel cortex of VIPcre/tdTomato mice in a layer-specific manner. We show that VIPcre/tdTomato mice are highly sensitive and specific for the entire population of VIP-expressing neurons. In the barrel cortex, approximately 13% of all GABAergic neurons are VIP expressing. Most VIP neurons are found in layer II/III (∼60%), whereas approximately 40% are found in the other layers of the barrel cortex. Layer II/III VIP neurons are significantly different from VIP neurons in layers IV-VI in several morphological and membrane properties, which suggest layer-dependent differences in functionality.
The neocortex is regarded as the brain structure responsible for mediating higher brain functions, like conscious perception of sensory signals, learning and memory or programming of goal-directed behavior. Cortical circuits that enable these functions are formed by, first, a larger population of excitatory so-called principal cells (i.e., glutamatergic pyramidal cells; ca. 80–85 %), which issue long-distance projections, in addition to local recurrent collaterals, which form the major part of local cortical excitatory circuits. A second, smaller population of inhibitory also called local or short-axoned interneurons (i.e., GABAergic neurons; ca. 15–20 %), however, contribute heavily to intracortical microcircuits too. They can be subdivided by their location in specific areas, layers, or columns, which possess specific input–output relationships, but also in terms of morphology, electrophysiology, molecular expression profiles, and subcellular target specificity. Here it is proposed that, at present, in the rodent neocortex this population of GABAergic neurons can be reasonably divided into six different types, mainly due to their unique axonal patterns and subcellular target specificity: (i) axo-axonic cells, (ii) basket cells, (iii) Martinotti cells, (iv) bipolar/bitufted cells, (v) neurogliaform cells, and (vi) projection neurons. These different types of GABAergic neurons strongly govern the working of cortical circuits for meaningful behavior by feed-forward and feedback inhibition as well as disinhibition. Thus, they keep excitation in check, perform gain modulation, and open temporal or spatial windows for input control or output generation.
Depolarizing chloride gradient in developing cochlear nucleus neurons: Underlying mechanism and implication for calcium signaling.
Witte M, Reinert T., Dietz B., Nerlich J., Rübsamen R., Milenkovic I..
Neuroscience 261: 207-222 , 2014.
Precise regulation of the chloride homeostasis crucially determines the action of inhibitory transmitters GABA and glycine and thereby endows neurons or even discrete neuronal compartments with distinct physiological responses to the same transmitters. In mammals, the signaling mediated by GABAA/glycine receptors shifts during early postnatal life from depolarization to hyperpolarization, due to delayed maturation of the chloride homeostasis system. While the activity of the secondary active, K+-Cl--extruding cotransporter KCC2, renders GABA/glycine hyperpolarizing in auditory brainstem nuclei of altricial rodents, the mechanisms contributing to the initially depolarizing transmembrane gradient for Cl- in respective neurons remained unknown. Here we used gramicidin-perforated patch recordings, non-invasive Cl- and Ca2+ imaging, and immunohistochemistry to identify the Cl--loading transporter that renders depolarizing effects of GABA/glycine in early postnatal life of spherical bushy cells in the cochlear nucleus of gerbil. Our data identify the 1Na+:1K+:2Cl- cotransporter 1 (NKCC1) as the major Cl--loader responsible for depolarizing action of GABA/glycine at postnatal days 3-5 (P3-5). Extracellular GABA/muscimol elicited calcium signaling through R-, L-, and T-type channels, which was dependent on bumetanide- and [Na+]e-sensitive Cl- accumulation. The "adult like", low intracellular Cl- concentration is established during the second postnatal week, through a mechanism engaging the NKCC1-down regulation between P5 and P15 and ongoing KCC2-mediated Cl--extrusion.
Presynaptic and postsynaptic origin of multicomponent extracellular spike waveforms at the endbulb of held/spherical bushy cell synapse..
Typlt M., Haustein M., Dietz B., Steinert J., Witte M., Englitz B., Milenkovic I., Kopp-Scheinpflug C., Forsythe I., Rübsamen R..
Eur J Neuroscience 31(9):1574-81 , 2010.
Extracellular signals from the endbulb of Held-spherical bushy cell (SBC) synapse exhibit up to three component waves ('P', 'A' and 'B'). Signals lacking the third component (B) are frequently observed but as the origin of each of the components is uncertain, interpretation of this lack of B has been controversial: is it a failure to release transmitter or a failure to generate or propagate an action potential? Our aim was to determine the origin of each component. We combined single- and multiunit in vitro methods in Mongolian gerbils and Wistar rats and used pharmacological tools to modulate glutamate receptors or voltage-gated sodium channels. Simultaneous extra- and intracellular recordings from single SBCs demonstrated a presynaptic origin of the P-component, consistent with data obtained with multielectrode array recordings of local field potentials. The later components (A and B) correspond to the excitatory postsynaptic potential (EPSP) and action potential of the SBC, respectively. These results allow a clear interpretation of in vivo extracellular signals. We conclude that action potential failures occurring at the endbulb-SBC synaptic junction largely reflect failures of the EPSP to trigger an action potential and not failures of synaptic transmission. The data provide the basis for future investigation of convergence of excitatory and inhibitory inputs in modulating transmission at a fully functional neuronal system using physiological stimulation.
P2 receptor-mediated signaling in spherical bushy cells of the mammalian cochlear nucleus..
Milenkovic, I., Rinke I, Witte M, Dietz, B., Rübsamen R..
J Neurophysiol. 102(3):1821-1833, 2009.
Purinoreceptors of the P2 family contribute strongly to signaling in the cochlea, but little is known about the effects of purinergic neurotransmission in the central auditory system. Here we examine P2 receptor-mediated signaling in the large spherical bushy cells (SBCs) of Mongolian gerbils around the onset of acoustically evoked signal processing (P9-P14). Brief adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS) application evoked inward current, membrane depolarization, and somatic Ca2+ signals. Moreover, ATPgammaS changed the SBCs firing pattern from phasic to tonic, when the application was synchronized with depolarizing current injection. This bursting discharge activity was dependent on [Ca2+]i and Ca2+-dependent protein kinase (PKC) activity and is presumably caused by modulation of low-threshold K+ conductance. Activation of P2Y1 receptors could not evoke these changes per se, thus it was concluded that the involvement of P2X receptors seems to be necessary. Ca2+ imaging data showed that both P2X and P2Y1 receptors mediate Ca2+ signals in SBCs where P2Y1 receptors most likely activate the PLC-IP3 (inositol trisphosphate) pathway and release Ca2+ from internal stores. Immunohistochemical staining confirmed the expression of P2X2 and P2Y1 receptor proteins in SBCs, providing additional evidence for the involvement of both receptors in signal transduction in these neurons. Purinergic signaling might modulate excitability of SBCs and thereby contribute to regulation of synaptic strength. Functionally, the increase in firing rate mediated by P2 receptors could reduce temporal precision of the postsynaptic firing, e.g., phase locking, which has an immediate effect on signal processing related to sound localization. This might provide a mechanism for adaptation to the ambient acoustic environment.
Development of chloride-mediated inhibition in neurons of the anteroventral cochlear nucleus of gerbil (Meriones unguiculatus).
Milenkovic I., Witte M., Turecek R., Heinrich M., Reinert T., Rübsamen R..
J Neurophysiol 98: 1634-1644, 2007.
At the initial stages in neuronal development, GABAergic and glycinergic neurotransmission exert depolarizing responses, assumed to be of importance for maturation, which in turn shift to hyperpolarizing in early postnatal life due to development of the chloride homeostasis system. Spherical bushy cells (SBC) of the mammalian cochlear nucleus integrate excitatory glutamatergic inputs with inhibitory (GABAergic and glycinergic) inputs to compute signals that contribute to sound localization based on interaural time differences. To provide a fundamental understanding of the properties of GABAergic neurotransmission in mammalian cochlear nucleus, we investigated the reversal potential of the GABA-evoked currents (E GABA) by means of gramicidin-perforated-patch recordings in developing SBC. The action of GABA switches from depolarizing to hyperpolarizing by the postnatal day 7 due to the negative shift in E GABA. Furthermore, we studied the expression pattern of the K+-Cl(-)-extruding cotransporter KCC2, previously shown to induce a switch from neonatal Cl(-) efflux to the mature Cl(-) influx in various neuron types, thereby causing a shift from depolarizing to hyperpolarizing GABA action. The KCC2 protein is expressed in SBC already at birth, yet its activity is attained toward the end of the first postnatal week as indicated by pharmacological inhibition. Interruption of the Cl(-) extrusion by [(dihydroindenyl)oxy] alkanoic acid or furosemide gradually shifted E(GABA) in positive direction with increasing maturity, suggesting that KCC2 could be involved in maintaining low [Cl(-)]i after the postnatal day 7 thereby providing the hyperpolarizing Cl(-)-mediated inhibition in SBC.