scholarly journals Inositol 1,4,5 trisphosphate releases calcium from specialized sites within Limulus photoreceptors

1987 ◽  
Vol 104 (4) ◽  
pp. 933-937 ◽  
Author(s):  
R Payne ◽  
A Fein
Keyword(s):  
Endoplasmic Reticulum ◽  
Injection Site ◽  
Intracellular Calcium ◽  
Subcellular Distribution ◽  
Smooth Endoplasmic Reticulum ◽  
Image Intensifier ◽  
Photoreceptor Cells ◽  
Calcium Stores ◽  
Inositol 1,4,5 Trisphosphate ◽  
Probable Site

We have investigated the subcellular distribution and identity of inositol trisphosphate (InsP3)-sensitive calcium stores in living Limulus ventral photoreceptor cells, where light and InsP3 are known to raise intracellular calcium. We injected ventral photoreceptor cells with the photoprotein aequorin and viewed its luminescence with an image intensifier. InsP3 only elicited detectable aequorin luminescence when injected into the light-sensitive rhabdomeral (R)-lobe where aequorin luminescence induced by light was also confined. Calcium stores released by light and InsP3 are therefore localized to the R-lobe. Within the R-lobe, InsP3-induced aequorin luminescence was further confined around the injection site, due to rapid dilution and/or degradation of injected InsP3. Prominent cisternae of smooth endoplasmic reticulum are uniquely localized within the cell beneath the microvillar surface of the R-lobe (Calman, B., and S. Chamberlain, 1982, J. Gen. Physiol., 80:839-862). These cisternae are the probable site of InsP3 action.

Imaging of intracellular calcium stores in single permeabilized lens cells

1999 ◽  
Vol 276 (2) ◽  
pp. C426-C434 ◽  
Author(s):  
Grant C. Churchill ◽  
Charles F. Louis
Keyword(s):  
Intracellular Calcium ◽  
Calcium Stores ◽  
Fura 2 ◽  
Intracellular Calcium Stores ◽  
Cyclic Adp Ribose ◽  
Inositol 1,4,5 Trisphosphate ◽  
Lens Cells ◽  
Intracellular Ca

Intracellular Ca2+ stores in permeabilized sheep lens cells were imaged with mag-fura 2 to characterize their distribution and sensitivity to Ca2+-releasing agents. Inositol 1,4,5-trisphosphate (IP3) or cyclic ADP-ribose (cADPR) released Ca2+ from intracellular Ca2+ stores that were maintained by an ATP-dependent Ca2+ pump. The IP3 antagonist heparin inhibited IP3- but not cADPR-mediated Ca2+ release, whereas the cADPR antagonist 8-amino-cADPR inhibited cADPR- but not IP3-mediated Ca2+ release, indicating that IP3 and cADPR were operating through separate mechanisms. A Ca2+ store sensitive to IP3, cADPR, and thapsigargin appeared to be distributed throughout all intracellular regions. In some cells a Ca2+ store insensitive to IP3, cADPR, thapsigargin, and 2,4-dinitrophenol, but not ionomycin, was present in a juxtanuclear region. We conclude that lens cells contain intracellular Ca2+ stores that are sensitive to IP3, cADPR, and thapsigargin, as well as a Ca2+store that appears insensitive to all these agents.

Hepatic Subcellular Compartmentation of Cytoplasmic Phosphoenolpyruvate Carboxykinase Determined by Immunogold Electron Microscopy

1995 ◽  
Vol 1 (4) ◽  
pp. 151-161
Author(s):  
Kuixiong Gao ◽  
Emma Lou Cardell ◽  
Randal E. Morris ◽  
Bruce F. Giffin ◽  
Robert R. Cardell
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Rat Liver ◽  
Subcellular Distribution ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Smooth Endoplasmic Reticulum ◽  
Immunogold Electron Microscopy ◽  
Immunogold Staining ◽  
Thin Sections ◽  
Gold Particles

Phosphoenolpyruvate carboxykinase (PEPCK) is the rate-limiting gluconeogenic enzyme and in liver occurs in a lobular gradient from periportal to pericentral regions. The subcellular distribution of cytoplasmic PEPCK molecules within hepatocytes and its relationship to organelles have not been determined previously. In this study, we have used immunogold electron microscopy to evaluate the subcellar distribution of the enzyme, in addition to brightfield and epipolarized light microscopy. Cryosections (10 μm) of perfusion-fixed rat liver were collected on silanated slides and immunostained using goat anti-rat PEPCK followed by 5-nm gold-labeled secondary and tertiary antibodies. Additionally, free-floating vibratome sections (25, 50, and 100 μm) of perfusion-immersion-fixed rat liver were immunogold stained using goat anti-rat PEPCK and 5-nm gold-labeled secondary antibody, with and without silver enhancement. The immunogold labeled sections from both procedures were embedded in epoxy resin for the preparation of thin sections for electron microscopy. The results showed that the gold-labeled antibodies penetrated the entire thickness of cryosections, resulting in a high signal for PEPCK, but membranes in general, the smooth endoplasmic reticulum in particular, were not identifiable as electron dense unit membranes. On the other hand, the vibratome sections of well-fixed tissue allowed good visualization of the ultrastructure of cellular organelles, with the smooth endoplasmic reticulum appearing as vesicles and tubules with electron dense unit membranes; however, the penetration of the gold-labeled antibody was limited to cells at the surface of the vibratome sections. In both procedures, PEPCK, as indicated by gold particles, is predominantly in the glycogen areas of the cytosome and not in mitochondria, nuclei, Golgi apparatus, or other cell organelles. Hepatocytes in periportal regions have a compact subcellular distribution of PEPCK shown by gold particles; hepatocytes in pericentral regions have a diffuse subcellular distribution of PEPCK and thus more scattered gold particles. When normal serum replaced the first antibody in the immunogold staining procedures, the background was very low.

scholarly journals Source and sinks for the calcium released during fertilization of single sea urchin eggs.

1985 ◽  
Vol 100 (5) ◽  
pp. 1522-1527 ◽  
Author(s):  
A Eisen ◽  
G T Reynolds
Keyword(s):  
Endoplasmic Reticulum ◽  
Intracellular Calcium ◽  
Sea Urchin ◽  
Image Intensifier ◽  
Metabolic Activation ◽  
Calcium Transient ◽  
Clear Zone ◽  
Calcium Store ◽  
Arbacia Punctulata ◽  
Mitochondrial Uncouplers

The source and sinks for the intracellular calcium released during fertilization were examined in single eggs from the sea urchin, Arbacia punctulata. Single eggs were microinjected with the calcium photoprotein, aequorin. The calcium-aequorin luminescence was measured with a microscope-photomultiplier or observed with a microscope-image intensifier-video system. In the normal egg a propagated release has been observed. The source of the calcium was investigated in the organelle-stratified centrifuged egg and by the use of mitochondrial uncouplers. In the organelle-stratified centrifuged egg, the calcium-aequorin luminescence was found to originate from the clear zone. The principal constituent of the clear zone is the endoplasmic reticulum. Other potential sources of calcium are the mitochondria. Their contribution to the calcium transient was investigated by exposure of aequorin-injected eggs to mitochondrial uncouplers either before or after fertilization. There was no calcium released from the mitochondria before fertilization. A very large calcium store was released from the mitochondria after fertilization. Interestingly, eggs fertilized in the presence of uncouplers showed no increase in the calcium-aequorin luminescence over untreated eggs. Apparently, in the absence of mitochondrial uptake, other sinks for calcium with affinity and capacity similar to the mitochondria exist, but their nature is unknown. We suggest that the endoplasmic reticulum is the source of the intracellular calcium released upon fertilization and that the mitochondria are the principal sink. The results are discussed with regard to the metabolic activation of the egg.

Organization of intracellular calcium regulatory proteins in the neuronal endoplasmic reticulum

1994 ◽  
Vol 52 ◽  
pp. 26-27
Author(s):  
Maryann E. Martone ◽  
Victoria M. Edelman ◽  
Saul A. Alba ◽  
Thomas J. Deerinck ◽  
Mark H. Ellisman
Keyword(s):  
Endoplasmic Reticulum ◽  
Intracellular Calcium ◽  
Dendritic Spines ◽  
Calcium Binding ◽  
Smooth Endoplasmic Reticulum ◽  
Membrane System ◽  
Regulatory Proteins ◽  
Purkinje Neurons ◽  
Storage Site ◽  
Ip3 Receptor

The smooth endoplasmic reticulum (SER) has been established as an intracellular calcium storage site in neurons. Although the SER appears to form a continuous membrane system within neurons, immunolocalization studies suggest that calcium regulatory proteins are not evenly distributed within the SER but are selectively concentrated or excluded from certain domains. The subcompartmenalization of the SER has been clearly demonstrated in Purkinje neurons where two proteins involved in the release of calcium from intracellular stores, the IP3 and ryanodine receptor, were differentially localized within dendrites. Both proteins were found associated with the SER in cell bodies and dendrites of chick Purkinje neurons but only labeling for the IP3 receptor was found within dendritic spines. A similar differential localization was described in Purkinje cell dendrites for the SER Ca++ATPase and calsequestrin, a lumenal calcium binding protein. The Ca++ATPase was found throughout dendrites and dendritic spines while calsequestrin was restricted to membranous profiles within the dendritic shaft.

A quantitative morphological study of the pineal organ in the goldfish, Carassius auratus

1981 ◽  
Vol 59 (7) ◽  
pp. 1312-1325 ◽  
Author(s):  
John A. McNulty
Keyword(s):  
Endoplasmic Reticulum ◽  
Pineal Organ ◽  
Smooth Endoplasmic Reticulum ◽  
Close Association ◽  
Photoreceptor Cells ◽  
Nerve Fibers ◽  
Substantial Part ◽  
Electron Microscopic ◽  
Goldfish Carassius Auratus ◽  
Supportive Cells

Stereological techniques applied to a light and electron microscopic study of the pineal organ of the goldfish indicated that photoreceptor and supportive cells were comparable in their number and cell volume and that approximately 500 nerve cells were present in the pineal end vesicle. There were approximately 310 nerve fibers descending the distal part of the pineal tract. Quantitative analysis of organelles in photoreceptor cells revealed that the endoplasmic reticulum and Golgi bodies, in the vicinity of which were situated both clear and dense-cored vesicles, formed a substantial part of the cytoplasmic volume. Other new observations reported for this species include a close association between mitochondria and parts of the smooth endoplasmic reticulum, a characteristic feature of photoreceptor cells, and the presence of subsurface cisternae formed from profiles of endoplasmic reticulum. Moreover, specialized contacts were found between both photoreceptor and supportive cells. Some of these ultrastructural features are similar to those reported in the secretory pinealocytes of mammals. These findings suggest that (1) the pineal organ in this species has a high degree of photosensitivity as evidenced by the large number of photoreceptor cells related to each nerve cell, and (2) photoreceptor cells are metabolically active possibly having functions other than photoreception.

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