Two Peeling Methods for the Isolation of Photoreceptor Cell Compartments in the Mouse Retina for Protein Analysis

10.3791/62977 ◽  
2021 ◽  
Author(s):  
Kasey Rose ◽  
Sowmya Lokappa ◽  
Jeannie Chen
Keyword(s):  
Photoreceptor Cell ◽  
Protein Analysis ◽  
Mouse Retina ◽  
Cell Compartments

scholarly journals Enhancer transcription identifies cis-regulatory elements for photoreceptor cell types

2019 ◽  
Author(s):  
Rangarajan D. Nadadur ◽  
Carlos Perez-Cervantes ◽  
Nicolas Lonfat ◽  
Linsin A. Smith ◽  
Andrew E. O. Hughes ◽  
...  
Keyword(s):  
Differential Expression ◽  
Regulatory Networks ◽  
Photoreceptor Cell ◽  
Cell Types ◽  
Regulatory Elements ◽  
Mouse Retina ◽  
Specific Cell ◽  
Rod Photoreceptor ◽  
Cone Photoreceptor ◽  
Accessible Chromatin

AbstractIdentification of the cis-regulatory elements (CREs) that regulate gene expression in specific cell types is critical for defining the gene regulatory networks (GRNs) that control normal physiology and disease states. We previously utilized non-coding RNA (ncRNA) profiling to define CREs that comprise a GRN in the adult mouse heart1. Here, we applied ncRNA profiling to the mouse retina in the presence and absence of Nrl, a rod photoreceptor-specific transcription factor required for rod versus cone photoreceptor cell fate. Differential expression of Nrl-dependent ncRNAs positively correlated with differential expression of Nrl-dependent local genes. Two distinct Nrl-dependent regulatory networks were discerned in parallel: Nrl-activated ncRNAs were enriched for accessible chromatin in rods but not cones whereas Nrl-repressed ncRNAs were enriched for accessible chromatin in cones but not rods. Furthermore, differential Nrl-dependent ncRNA expression levels quantitatively correlated with photoreceptor cell type-specific ATAC-seq read density. Direct assessment of Nrl-dependent ncRNA-defined loci identified functional cone photoreceptor CREs. This work supports differential ncRNA profiling as a platform for identifying context-specific regulatory elements and provides insight into the networks that define photoreceptor cell types.

Long-term ERG analysis in the partially light-damaged mouse retina reveals regressive and compensatory changes

2006 ◽  
Vol 23 (1) ◽  
pp. 91-97 ◽  
Author(s):  
ADAM RICHARDS ◽  
ALFRED A. EMONDI ◽  
BAERBEL ROHRER
Keyword(s):  
Photoreceptor Cell ◽  
Light Exposure ◽  
Cell Loss ◽  
Oscillatory Potentials ◽  
Mouse Retina ◽  
Dependent Manner ◽  
Light Damage ◽  
Outer Retina ◽  
Inner Retina ◽  
Baseline Values

Most of the blinding retinopathies are due to progressive photoreceptor degeneration. Treatment paradigms that are currently being investigated include strategies to either halt or slow down photoreceptor cell loss, or to replace useful vision with retinal prosthesis. However, more information is required on the pathophysiological changes of the diseased retina, in particular the inner retina, that occur as a consequence of photoreceptor cell loss. Here we wished to use light damage as a stoppable insult to determine the structural and functional consequences on inner and outer retina, with the overall goal of determining whether survival of a functional inner retina is possible even if the outer retina is damaged. Mice were exposed to a 20-day light-damage period. Electroretinograms (ERG) and morphology were used to assess subsequent recovery. Outer retina was monitored analyzinga-waves, which represent photoreceptor cell responses, and histology. Integrity of the inner retina was monitored, analyzingb-waves and oscillatory potentials (OP1–OP4) and immunohistochemical markers for known proteins of the inner retina. All six ERG components were significantly suppressed with respect to amplitudes and kinetics, but stabilized in a wave-dependent manner within 40–70 days after the end of light exposure. As expected, damage of the outer retina was permanent. However, function of the inner retina was found to recover significantly. Whileb-wave amplitudes remained suppressed to 60% of their baseline values, OP amplitudes recovered completely, and implicit times of all components of the inner retina (b-wave and OP1–OP4) recovered to a level close to baseline values. Histological analyses confirmed the lack of permanent damage to the inner retina. In summary, these data suggests that the inner retina has the potential for significant recovery as well as plasticity if treatment is available to stop the deterioration of the outer retina.

STAT3-Mediated Signaling in the Determination of Rod Photoreceptor Cell Fate in Mouse Retina

2004 ◽  
Vol 45 (7) ◽  
pp. 2407 ◽  
Author(s):  
Samuel Shao-Min Zhang ◽  
Jiye Wei ◽  
Hua Qin ◽  
Lixin Zhang ◽  
Bing Xie ◽  
...  
Keyword(s):  
Cell Fate ◽  
Photoreceptor Cell ◽  
Mouse Retina ◽  
Rod Photoreceptor

Ionic homeostasis and subcellular element compartmentation

1990 ◽  
Vol 48 (2) ◽  
pp. 150-151
Author(s):  
Ann LeFurgey ◽  
Peter Ingram ◽  
J.J. Blum ◽  
M.C. Carney ◽  
L.A. Hawkey ◽  
...  
Keyword(s):  
Leishmania Major ◽  
Ionic Homeostasis ◽  
X Ray ◽  
Functional Studies ◽  
Cell Compartments ◽  
X Ray Imaging ◽  
Resolution Electron ◽  
Multiple Cell ◽  
Role Of Zn

Subcellular compartments commonly identified and analyzed by high resolution electron probe x-ray microanalysis (EPXMA) include mitochondria, cytoplasm and endoplasmic or sarcoplasmic reticulum. These organelles and cell regions are of primary importance in regulation of cell ionic homeostasis. Correlative structural-functional studies, based on the static probe method of EPXMA combined with biochemical and electrophysiological techniques, have focused on the role of these organelles, for example, in maintaining cell calcium homeostasis or in control of excitation-contraction coupling. New methods of real time quantitative x-ray imaging permit simultaneous examination of multiple cell compartments, especially those areas for which both membrane transport properties and element content are less well defined, e.g. nuclei including euchromatin and heterochromatin, lysosomes, mucous granules, storage vacuoles, microvilli. Investigations currently in progress have examined the role of Zn-containing polyphosphate vacuoles in the metabolism of Leishmania major, the distribution of Na, K, S and other elements during anoxia in kidney cell nuclel and lysosomes; the content and distribution of S and Ca in mucous granules of cystic fibrosis (CF) nasal epithelia; the uptake of cationic probes by mltochondria in cultured heart ceils; and the junctional sarcoplasmic retlculum (JSR) in frog skeletal muscle.

Protein analysis of pediatric parapneumonic pleural effusion after intrapleural urokinase treatment

2010 ◽  
Vol 34 (8) ◽  
pp. S69-S69
Author(s):  
Jieh‑Neng Wang ◽  
Pao‑Chi Liao ◽  
Yu‑Chin Tasi ◽  
Jing‑Ming Wu
Keyword(s):  
Pleural Effusion ◽  
Protein Analysis ◽  
Parapneumonic Pleural Effusion
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