Primary Lysosomes

L Auer rods are abnormal lysosomes (primary granules) that are pathognomonic of myeloblasts and not found in ALL. 

Dilute primary antibody in 1% BSA in 0.1% Triton X-IOO/PBS. Rat anti-CXCR4 (1 100) in combination with mouse anti-EEAl (1 1000) or anti-LAMPl (1 1000). EEAl is used as a marker for early endosomes and LAMP2 is used as a marker for late endosomes/lysosomes. Primary antibody titrations should be performed to identify optimal antibody dilutions to use for staining. 

Fig. 7. Transport pathways of a polymer in the liver 1 — diffusion through theintercellular junctions (molecular-size limited process) 2 — transcellular route of polymer transport into the bile including pinocytosis into hqtatocyte and exocytosis of the vesicles or residual bodies at the lateral side of the cell 3 — pinocytosis into the Kupffer cells occurs regularly, from either the central or the interstitial compartment. H.C. — hepatocyte E.C. — endothelial cell of capillary wall K.C. — Kupffer cell I.S. — interstitial space B.C. — bile canaliculi P — pinocytic vesicle L — lysosomes primary) S.L. — secondary lysosome R.B. — residual body N — nudeus...

FIGURE 1-6 A portion of a Golgi apparatus. The smooth-mem-braned cisternae appear beaded. The many circular profiles represent tangentially sectioned fenestrations and alveolate vesicles (primary lysosomes). Two of the latter can be seen budding from Golgi saccules (arrows). Mitochondria and a dense body (secondary lysosomes) are also present. x60,000. 

Multivesicular bodies are usually found in association with the Golgi apparatus and are visualized by EM as small, single membrane-bound sacs approximately 0.5 Jim in diameter. They contain several minute, spherical profiles, sometimes arranged about the periphery. They are believed to belong to the lysosome series prior to secondary lysosomes because they contain acid hydrolases and apparently are derived from primary lysosomes. 

Primary lysosomal hydrolase defects 685 Other types of lysosomal disorder 688... 

Primary lysosomal hydrolase defects. Two-thirds of the lysosomal storage diseases involve defects in genes that code for acid hydrolases. Table 41-2 lists 29 defects that have been defined so far. They have an autosomal recessive mode of inheritance, except for Hunter s syndrome and Fabry s disease, where the mode is X-linked recessive. The defective genes have been identified and mutations have been defined for nearly all. The nervous system is involved in most. Many of the disorders show a wide range of clinical severity, which may range from death in early childhood to a moderate disability in adulthood. 

In microorganisms, visualization of the lysosomal system of different species of malaria parasites, (19), the endoplasmic reticulum-Golgi complex system of Tritrichomonas foetus (20) and the Golgi complex and primary lysosomes of Pneumocystis carinii (30) has been attempted using ZIO staining. 

Kagedal, K., Bironaite, D., and Olhnger, K., 1999, Anthraquinone cytotoxicity and apoptosis in primary cultures of hepatocytes. Free Rad. Res. 31 419-428 Kagedal, K., Johansson, U., and Olhnger, K., 2001, The lysosomal protease cathepsin D mediates apoptosis induced by oxidative stress. FASEB J. (May 18, 2001) 10.1096/. 00-0708fje... 

Monney, L., Ohvier, R., Otter, 1., Jansen, B., Poirier, GG., and Bomer, C., 1998, Role ofan acidic compartment in tumor-necrosis-factor-a-induced production of ceramide, activation of caspase-3 and apoptosis. Eur. J. Biochem. 251 295-303 Morrison, H., Jemstrom, B., Nordenskjdld, M., Thor, H., and Orrenius, S., 1984, Induction of DNA damage by menadione (2-methyl-l,4-naphthoquinone) in primary cultures of rat hepatocytes. Biochem. Pharmacol. 33 1763-1769 Neuzil, J., Svensson, 1., Weber, T, Weber, C., and Brunk, U.T., 1999, alpha-tocopheryl succinate-induced apoptosis in Jurkat T cells involves caspase-3 activation, and both lysosomal and mitochondrial destabilisation. FEES Lett. 445 295-300 Ngo, E.O., Nutter, L.M., Sura, T., and Gutierrez, P. L., 1998, Induction ofp53 by the... 

Figure 1.6 Vesicular transport of proteins within the cell. Vesicles from the endoplasmic reticulum [A] carry protein to the Golgi complex, they are repackaged in the Golgi from which they leave to form primary lysosomes [B] or fuse with the plasma membrane this is to add proteins or to be secreted from the cell [C]. In the Golgi, new vesicles are formed to transport the proteins to the plasma membrane (e.g. transport proteins or proteins for export) or the lysosomes. This system transports, safely, dangerous hydrolytic enzyme to the lysosomes and it also protects membrane proteins, or proteins for export, from degradation in the cytosol.

Drugs that have primary amino groups available for conjugation, for instance dopamine and doxorubicin, can in principle be coupled to LMWPs via oligopeptides. In contrast to the carboxypeptidases, the aminopeptidases appear to possess a broader specificity. To allow the release of terminal amino group-containing drugs in the acid environment of the lysosomes without the requirement of enzymes, an acid-sensitive spacer can be used. 

The membranes of the Golgi apparatus contain receptor molecules that bind Man 6-P. They recognize lysosomal proproteins by this residue and bind them (3). With the help of clathrin, the receptors are concentrated locally. This allows the appropriate membrane sections to be pinched off and transported to the endolysosomes with the help of transport vesicles (4), from which primary lysosomes arise through maturation (5). Finally, the phosphate groups are removed from Man 6-P (6). 

Fig. I. Endocytic pathways used by cells to internalize soluble macromolecules [25] fluid-phase pinocytosis (1), adsorptive pinocytosis (2), and receptor-mediated endocytosis (pinocytosis) (6). Each of these processes involves a formation of a sealed vesicle formed from the plasma membrane which encloses part of the extracellular medium. The internalization of a polymer-drug conjugate (P-D), and targeted polymer-drug conjugate ( => —P-D) is shown. Other abbreviations — = cell surface receptor/antigen 1 = clathrin molecule X = lysosomal enzyme. Fluid-phase pinocytosis (1) and adsorptive pinocytosis (2) are nonspecific processes which direct the macromolecule into the lysosomal compartment of the cell. Once P-D is internalized, whether by (1) or (2), the resulting endosome (3) is ultimately fused with a primary lysosome (4) forming a secondary lysosome (5). In the latter compartment P-D is in contact with several types of lysosomal enzymes. The membrane of (5) is impermeable to macromolecules. Consequently, the structure of P-D may be designed in such...

There are four main compartments a soluble macromolecule can enter the central compartment (blood and lymphatic system), interstitium, intestinal lumen, and lysosomes [100, 101]. Minor compartments are primary urine, liquor, bile, etc. There is no experimental evidence that clearly indicates the penetration of synthetic macromolecules into the cytoplasm, i.e, into the intracellular compartment (inside the cell but outside the endosomes or lysosomes) [101]. The movements of soluble macromolecules between body compartments have been extensively reviewed [14, 20,100-104] and will not be covered in detail here. We shall concentrate on the discussion of main factors influencing the movement of soluble macromolecules when administered into the bloodstream. Depending on the structure and molecular weight distribution, part of the polymeric molecules are excreted in the urine. Simultaneously, the macromolecules are cleared from the bloodstream by endocytosis. It is important to note that nonspecific capture of soluble macromolecules by the specialized cells of the reticuloendothelial system is generally much less (orders of magnitude) when compared to vesicular carriers of a comparable structure. 

Secondary events result from primary events, for example, changes in membrane structure/permeability, mitochondrial damage, and lysosomal destabilization. Tertiary events are final observable manifestations, for example, fatty change and phospholipidosis, apoptosis, blebbing, and necrosis. 

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