Inner limiting membrane

Figure 14.1 Schematic diagram of the blood-retinal barrier (BRB). The retinal cell layers seen histologically consist of retinal pigment epithelium (RPE) photoreceptor outer segments (POS) outer limiting membrane (OLM) outer nuclear layer (ONL) outer plexiform layer (OPL) inner nuclear layer (INL) inner plexiform layer (IPL) ganglion cell layer (GCL) nerve fiber layer (NFL) inner limiting membrane (ILM).

Inner limiting membrane Ganglion cell layer Inner pi ex i form layer... 

Therefore it appears that we can achieve and maintain intraocular levels of either PEDF or K1K3 angiostatin in neonatal and adult mice sufficient to expect significant reduction of retinal NV. In the ischemic mouse model the level of retinal NV is measured quantitatively by enumerating the endothelial cells above the inner limiting membrane (ILM) of the retina (see later). Such an analysis showed that PEDF treated eyes had 74% fewer endothelial cells above the ILM compared to paired controls and 78% fewer compared to paired controls for K1K3 treated eyes (Raisler et al., 2002). 

The ganglion cell layer (GCL) contains the cell bodies of retinal ganglion cells, with their axons running across the retinal surface (nerve fiber layer) toward the optic nerve head, and on through the optic nerve to the lateral geniculate nucleus in the mid-brain. The inner retinal blood supply (outside the foveal avascular zone), the nerve fiber layer, and a thin membrane (the inner limiting membrane) form the most superficial retinal structures. 

The inner limiting membrane formed by the endfeet of Muller cells. 

Most of the NADH used in electron transport is produced in the mitochondrial matrix space, an appropriate site because NADH is oxidized by Complex I on the matrix side of the inner membrane. Furthermore, the inner mitochondrial membrane is impermeable to NADH. Recall, however, that NADH is produced in glycolysis by glyceraldehyde-3-P dehydrogenase in the cytosol. If this NADH were not oxidized to regenerate NAD, the glycolytic pathway would cease to function due to NAD limitation. Eukaryotic cells have a number of shuttle systems that harvest the electrons of cytosolic NADH for delivery to mitochondria without actually transporting NADH across the inner membrane (Figures 21.33 and 21.34). 

In addition to the processes described above, there still remains one further process which, at least in some cells or tissues, is required prior to the utilisation of ATP in the cytosol that is, the transport of energy within the cytosol, via a shuttle. The transport of ATP out and ADP into the mitochondrion, via the translocase, results in a high ATP/ ADP concentration ratio in the cytosol. However, a high ratio means that the actual concentration of ADP in the cytosol is low, which could result in slow diffusion of ADP from a site of ATP utilisation back to the inner mitochondrial membrane. If sufficiently slow, it could limit the rate of ATP generation. To overcome this, a process exists that transports energy within the cytosol, not by diffusion of ATP and ADP, but by the diffusion of phosphocreatine and creatine, a process known as the phosphocreatine/creatine shuttle. The reactions involved in the shuttle in muscle help to explain the significance of the process. They are ... 

The inner surface of the silicalemma, i.e., the limiting membrane within which silica deposition occurs549 consists of a protein template enriched in serine and threonine. This protein will present a layer of hydroxyl groups which can undergo condensation reaction with silicic acid molecules with a consequent loss of water (Fig. 48). As a result, the initial layer of condensed silicic acid will be held fixed to the protein template of silicic acid (Fig. 49). Such a situation is kinetically more favorable than simply allowing the silicic acid molecules to come together by random collision. 

ADP phosphorylation is tightly coupled to electron transport. Shutting down one shuts down the other. It is well known that if ADP phosphorylation is inhibited by such compounds as oligomycin, electron transport also ceases. If the proton gradient is broken by a proton ionophore, however, such as 2,4-dinitrophenol, electron transport resumes at a rapid pace and no phosphorylation takes place. Such proton ionophores are also termed "uncouplers" of electron transport and ADP phosphorylation. Under normal conditions, the factors limiting ATP production are the pH gradient across the inner mitochondrial membrane and the cellular ADP/ATP ratio. An increase in the proton gradient shuts down phosphorylation and electron transport, whereas an increase in the ADP/ATP ratio stimulates both. Stimulation of oxidative phosphorylation by increases in cellular ADP concentration is termed respiratory control. 

Fig. 3. The rate-limiting step of steroidogenesis under ACTH regulation. The transfer of cholesterol (C) from the outer to the inner mitochondrial membrane under ACTH regulation (step 3) makes cholesterol available to cytochrome /M50scc for conversion to pregnenolone (step 4), which diffuses out of the mitochondrion (step 5). Because of its insolubility in aqueous media, cholesterol must be transported to mitochondria, probably by SCP2, from a precursor pool (step 2). Here, cholesterol in the precursor pool is shown as being formed from cholesterol esters (CE) by cholesterol ester hydrolase (CEH) (step 1) other possible pathways are shown in Figs. 4 and 6. From Ref. 14.

Phosphorylation of ADP to ATP by mitochondria is driven by an electrochemical proton gradient established across the inner mitochondrial membrane as a consequence of vectoral transport of protons from NADH and succinate during oxidation by the respiratory chain (see Chapter 17). Hence, lipophilic weak acids or bases (such as 2,4-dinitrophenol) that can shuttle protons across membranes will dissipate the proton gradient and uncouple oxidation from ADP phosphorylation. Intrami-tochondrial ADP can be rate-limiting as demonstrated by inhibition of the mitochondrial adenosine nucleotide carrier by atractyloside. Inhibition of ATP synthesis... 

The need for energy by the cell regulates the TCA cycle, which acts in concert with the electron transfer chain and the ATPase to produce ATP in the inner mitochondrial membrane. The cell has limited amounts of ATP, ADP,... 

Inside the inner membrane of a mitochondrion is a viscous region known as the matrix (Fig. 1-9). Enzymes of the tricarboxylic acid (TCA) cycle (also known as the citric acid cycle and the Krebs cycle), as well as others, are located there. For substrates to be catabolized by the TCA cycle, they must cross two membranes to pass from the cytosol to the inside of a mitochondrion. Often the slowest or rate-limiting step in the oxidation of such substrates is their entry into the mitochondrial matrix. Because the inner mitochondrial membrane is highly impermeable to most molecules, transport across the membrane using a carrier or transporter (Chapter 3, Section 3.4A) is generally invoked to explain how various substances get into the matrix. These carriers, situated in the inner membrane, might shuttle important substrates from the lumen between the outer and the inner mitochondrial membranes to the matrix. Because of the inner membrane, important ions and substrates in the mitochondrial matrix do not leak out. Such permeability barriers between various subcellular compartments improve the overall efficiency of a cell. 

Accompanying electron flow in mitochondria, H+ is transported from the matrix side of the inner membrane to the lumen between the limiting membranes, i.e., within the cristae (Figs. 1-9 and 6-9). Certain electron flow components are situated in the membranes such that they can carry out this vectorial movement. Protein Complex I, which oxidizes NADH, apparently transfers four H+ s across the inner membrane per pair of electrons from NADH. Complex II, which oxidizes FADH2 and leads to the reduction of a ubiquinone whose two electrons move to Complex III, apparently causes no H+ s to move from the matrix to the lumen. Transport of four H+ s from the matrix to the lumen side most likely occurs through protein Complex III per pair of electrons traversing the electron transport chain. Complex IV (cytochrome oxidase) may also transport four H+ s (Fig. 6-9 summarizes these possibilities). We also note that two H+ s are necessary for the reduction of 02 to H20, and these protons can also be taken up on the matrix side (Fig. 6-9). 

Conversion of cholesterol to pregnenolone is the rale-limiting step in steroid hormone biosynthesis. It is not ihccnry-malic transformation itself that is rate limiting, however, the translocation of cholesterol to the inner milochondrui membrane of steroid-synihesi/.ing cells is rate limiting. A key protein involved in the translocation is the 5Kruidugcmc... 

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