scholarly journals Targeted two-photon chemical apoptotic ablation of defined cell types in vivo

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Robert A. Hill ◽  
Eyiyemisi C. Damisah ◽  
Fuyi Chen ◽  
Alex C. Kwan ◽  
Jaime Grutzendler
Keyword(s):  
Cell Types ◽  
Two Photon

scholarly journals Distinct features of brain perivascular fibroblasts and mural cells revealed by in vivo two-photon imaging

2021 ◽  
pp. 0271678X2110685
Author(s):  
Stephanie K Bonney ◽  
Liam T Sullivan ◽  
Timothy J Cherry ◽  
Richard Daneman ◽  
Andy Y Shih
Keyword(s):  
Cell Types ◽  
Structural Features ◽  
Main Trunk ◽  
Perivascular Niche ◽  
Two Photon ◽  
Mural Cells ◽  
Adult Mice ◽  
Photon Imaging ◽  
Two Photon Imaging

Perivascular fibroblasts (PVFs) are recognized for their pro-fibrotic role in many central nervous system disorders. Like mural cells, PVFs surround blood vessels and express Pdgfrβ. However, these shared attributes hinder the ability to distinguish PVFs from mural cells. We used in vivo two-photon imaging and transgenic mice with PVF-targeting promoters (Col1a1 or Col1a2) to compare the structure and distribution of PVFs and mural cells in cerebral cortex of healthy, adult mice. We show that PVFs localize to all cortical penetrating arterioles and their offshoots (arteriole-capillary transition zone), as well as the main trunk of only larger ascending venules. However, the capillary zone is devoid of PVF coverage. PVFs display short-range mobility along the vessel wall and exhibit distinct structural features (flattened somata and thin ruffled processes) not seen with smooth muscle cells or pericytes. These findings clarify that PVFs and mural cells are distinct cell types coexisting in a similar perivascular niche.

scholarly journals Chronic imaging of mitochondria in the murine cerebral vasculature using in vivo two-photon microscopy

2020 ◽  
Vol 318 (6) ◽  
pp. H1379-H1386
Author(s):  
Ibolya Rutkai ◽  
Wesley R. Evans ◽  
Nikita Bess ◽  
Tomas Salter-Cid ◽  
Siniša Čikić ◽  
...  
Keyword(s):  
Cerebral Circulation ◽  
Three Dimensional ◽  
Cell Types ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Dimensional Reconstruction ◽  
Different Cell Types

We introduce an innovative in vivo approach to study mitochondria in the cerebral circulation in their physiological environment by demonstrating the feasibility of long-term imaging and three-dimensional reconstruction. We postulate that the appropriate combination of Cre/Lox system and two-photon microscopy will contribute to a better understanding of the role of mitochondria in not only endothelium but also the different cell types of the cerebral circulation.

scholarly journals Temporal dynamics of inhalation-linked activity across defined subpopulations of mouse olfactory bulb neurons imaged in vivo

2019 ◽  
Author(s):  
Shaina M. Short ◽  
Matt Wachowiak
Keyword(s):  
Olfactory Bulb ◽  
Temporal Dynamics ◽  
Cell Types ◽  
Response Patterns ◽  
Two Photon ◽  
Cell Type Specific ◽  
Olfactory Processing ◽  
Tufted Cells ◽  
Cell Subpopulations

ABSTRACTIn mammalian olfaction, inhalation drives the temporal patterning of neural activity that underlies early olfactory processing, and a single inhalation of odorant is sufficient for odor perception. However, how the neural circuits that process incoming olfactory information are activated in the context of inhalation-linked dynamics remains poorly understood. To better understand early olfactory processing in vivo, we used an artificial inhalation paradigm combined with two-photon calcium imaging to compare the dynamics of activity evoked by odorant inhalation across major cell types of the mouse olfactory bulb. Transgenic models and cell-type specific genetic tools were used to express GCaMP6f or jRGECO1a in mitral and tufted cell subpopulations, olfactory sensory neurons and two major juxtaglomerular interneuron classes, and responses to a single inhalation of odorant were compared. Activity in all cell types was strongly linked to inhalation, and all cell types showed some variance in the latency, rise-times and durations of their inhalation-linked response patterns. The dynamics of juxtaglomerular interneuron activity closely matched that of sensory neuron inputs, while mitral and tufted cells showed the highest diversity in dynamics, with a range of latencies and durations that could not be accounted for by heterogeneity in the dynamics of sensory input. Surprisingly, temporal response patterns of mitral and superficial tufted cells were highly overlapping such that these two subpopulations could not be distinguished on the basis of their inhalation-linked dynamics, with the exception of a subpopulation of superficial tufted cells expressing the peptide transmitter cholecystokinin. Overall, these results support a model in which diversity in inhalation-linked patterning of OB output arises first at the level of OSN inputs to the OB and is enhanced by feedforward inhibition from juxtaglomerular interneurons which differentially impacts different subpopulations of OB output neurons.

scholarly journals Distinct features of brain perivascular fibroblasts and mural cells revealed by in vivo two-photon imaging

2021 ◽  
Author(s):  
Stephanie K Bonney ◽  
Liam T Sullivan ◽  
Timothy Joel Cherry ◽  
Richard Daneman ◽  
Andy Y Shih
Keyword(s):  
Cell Types ◽  
Structural Features ◽  
Main Trunk ◽  
Perivascular Niche ◽  
Two Photon ◽  
Mural Cells ◽  
Adult Mice ◽  
Photon Imaging ◽  
Two Photon Imaging

Perivascular fibroblasts (PVFs) are recognized for their pro-fibrotic role in many central nervous system disorders. Like mural cells, PVFs surround blood vessels and express Pdgfrβ. However, these shared attributes hinder the ability to distinguish PVFs from mural cells. We used in vivo two-photon imaging and transgenic mice with PVF-targeting promoters (Col1a1 or Col1a2) to compare the structure and distribution of PVFs and mural cells in cerebral cortex of healthy, adult mice. We show that PVFs localize to all cortical penetrating arterioles and their pre-capillary offshoots, as well as the main trunk of only larger ascending venules. However, the capillary zone is devoid of PVF coverage. PVFs display short-range mobility along the vessel wall and exhibit distinct structural features (flattened somata and thin ruffled processes) not seen with smooth muscle cells or pericytes. These findings clarify that PVFs and mural cells are distinct cell types coexisting in a similar perivascular niche.

scholarly journals Developmental Divergence of Sensory Stimulus Representation in Cortical Interneurons

2020 ◽  
Author(s):  
Rahel Kastli ◽  
Rasmus Vighagen ◽  
Alexander van der Bourg ◽  
Ali Ozgur Argunsah ◽  
Asim Iqbal ◽  
...  
Keyword(s):  
Barrel Cortex ◽  
Cell Types ◽  
Thalamic Nuclei ◽  
Two Photon ◽  
Stimulus Representation ◽  
Adult Mice ◽  
Cortical Interneurons ◽  
Direct Input ◽  
Developmental Divergence

AbstractTwo inhibitory cell types involved in modulating barrel cortex activity and perception during active whisking in adult mice, are the VIP+ and SST+ interneurons. Here we identify a developmental transition point of structural and functional rearrangements onto these interneuron types around the start of active sensation at P14. Using in vivo two-photon Ca2+ imaging, we find that before P14, both interneuron types respond stronger to a multi-whisker stimulus, whereas after P14 their responses diverge, with VIP+ cells losing their multi-whisker preference and SST+ neurons enhancing theirs. Rabies virus tracings followed by tissue clearing, as well as photostimulation-coupled electrophysiology reveal that SST+ cells receive higher cross-barrel inputs compared to VIP+ at both time points. In addition, we also uncover that whereas prior to P14 both cell types receive direct input from the sensory thalamus, after P14 VIP+ cells show reduced inputs and SST+ cells largely shift to motor-related thalamic nuclei.

scholarly journals NaNuTrap, a technique for in vivo cell nucleus labelling using nanobodies

Development ◽  
2021 ◽  
Author(s):  
Zsuzsa Ákos ◽  
Leslie Dunipace ◽  
Angelike Stathopoulos
Keyword(s):  
Fluorescent Proteins ◽  
Cell Types ◽  
Cell Counting ◽  
Cell Labelling ◽  
Cell Nuclei ◽  
Maturation Time ◽  
Developmental Processes ◽  
Fluorescent Signal ◽  
Two Photon

In vivo cell labelling is challenging in fast developmental processes because many cell types differentiate more quickly than the maturation time of fluorescent proteins making visualization of these tissues impossible with standard techniques. Here we present a nanobody-based method, Nanobody Nuclear Trap (NaNuTrap), which works with the existing Gal4/UAS system in Drosophila and allows for early in vivo cell nuclei labelling independent of the fluorescent protein's maturation time. This restores the utility of fluorescent proteins that have longer maturation times, such as those used in two-photon imaging, for live imaging of fast or very early developmental processes. We also present a more general application of this system, whereby NaNuTrap can convert cytoplasmic GFP expressed in any existing transgenic fly line into a nuclear label. This nuclear re-localization of the fluorescent signal can improve the utility of the GFP label, for example in cell counting, as well as resulting in a general increase in intensity of the live fluorescent signal. We demonstrate these capabilities of NaNuTrap by effectively tracking subsets of cells during the fast movements associated with gastrulation.

scholarly journals Developmental divergence of sensory stimulus representation in cortical interneurons

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Rahel Kastli ◽  
Rasmus Vighagen ◽  
Alexander van der Bourg ◽  
Ali Özgür Argunsah ◽  
Asim Iqbal ◽  
...  
Keyword(s):  
Barrel Cortex ◽  
Cell Types ◽  
Thalamic Nuclei ◽  
Two Photon ◽  
Stimulus Representation ◽  
Signaling Dynamics ◽  
Cortical Interneurons ◽  
Direct Input ◽  
Developmental Divergence

AbstractVasocative-intestinal-peptide (VIP+) and somatostatin (SST+) interneurons are involved in modulating barrel cortex activity and perception during active whisking. Here we identify a developmental transition point of structural and functional rearrangements onto these interneurons around the start of active sensation at P14. Using in vivo two-photon Ca2+ imaging, we find that before P14, both interneuron types respond stronger to a multi-whisker stimulus, whereas after P14 their responses diverge, with VIP+ cells losing their multi-whisker preference and SST+ neurons enhancing theirs. Additionally, we find that Ca2+ signaling dynamics increase in precision as the cells and network mature. Rabies virus tracings followed by tissue clearing, as well as photostimulation-coupled electrophysiology reveal that SST+ cells receive higher cross-barrel inputs compared to VIP+ neurons at both time points. In addition, whereas prior to P14 both cell types receive direct input from the sensory thalamus, after P14 VIP+ cells show reduced inputs and SST+ cells largely shift to motor-related thalamic nuclei.

In vivo spectral imaging of different cell types in the small intestine by two-photon excited autofluorescence

2011 ◽  
Vol 16 (11) ◽  
pp. 116025 ◽  
Author(s):  
Regina Orzekowsky-Schroeder ◽  
Antje Klinger ◽  
Björn Martensen ◽  
Maike Blessenohl ◽  
Andreas Gebert ◽  
...  
Keyword(s):  
Small Intestine ◽  
Spectral Imaging ◽  
Cell Types ◽  
Two Photon ◽  
Different Cell Types

scholarly journals Versatile Surface Electrodes for Combined Electrophysiology and Two-Photon Imaging of the Mouse Central Nervous System

2021 ◽  
Vol 15 ◽  
Author(s):  
Michael Schweigmann ◽  
Laura C. Caudal ◽  
Gebhard Stopper ◽  
Anja Scheller ◽  
Klaus P. Koch ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Imaging Techniques ◽  
Brain Activity ◽  
Cell Types ◽  
High Signal ◽  
Neural Function ◽  
Two Photon ◽  
Cranial Window ◽  
Additional Stabilization

Understanding and modulating CNS function in physiological as well as pathophysiological contexts remains a significant ambition in research and clinical applications. The investigation of the multifaceted CNS cell types including their interactions and contributions to neural function requires a combination of the state-of-the-art in vivo electrophysiology and imaging techniques. We developed a novel type of liquid crystal polymer (LCP) surface micro-electrode manufactured in three customized designs with up to 16 channels for recording and stimulation of brain activity. All designs include spare central spaces for simultaneous 2P-imaging. Nanoporous platinum-plated contact sites ensure a low impedance and high current transfer. The epidural implantation of the LCP micro-electrodes could be combined with standard cranial window surgery. The epidurally positioned electrodes did not only display long-term biocompatibility, but we also observed an additional stabilization of the underlying CNS tissue. We demonstrate the electrode’s versatility in combination with in vivo 2P-imaging by monitoring anesthesia-awake cycles of transgenic mice with GCaMP3 expression in neurons or astrocytes. Cortical stimulation and simultaneous 2P Ca2+ imaging in neurons or astrocytes highlighted the astrocytes’ integrative character in neuronal activity processing. Furthermore, we confirmed that spontaneous astroglial Ca2+ signals are dampened under anesthesia, while evoked signals in neurons and astrocytes showed stronger dependency on stimulation intensity rather than on various levels of anesthesia. Finally, we show that the electrodes provide recordings of the electrocorticogram (ECoG) with a high signal-to noise ratio and spatial signal differences which help to decipher brain activity states during experimental procedures. Summarizing, the novel LCP surface micro-electrode is a versatile, convenient, and reliable tool to investigate brain function in vivo.

Ultrastructural Study of a differentiating human colon carcinoma cell line

1990 ◽  
Vol 48 (3) ◽  
pp. 208-209
Author(s):  
Sylvie Polak-Charcon ◽  
Mehrdad Hekmati ◽  
Yehuda Ben Shaul
Keyword(s):  
Cell Line ◽  
Morphological Changes ◽  
Secretory Granules ◽  
Human Colon ◽  
Cell Types ◽  
Culture Conditions ◽  
Morphological Evidence ◽  
Human Colon Carcinoma Cell ◽  
Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

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