Biomedical subjects

U Schraermeyer

Publications and source records attributed to U Schraermeyer.

At least 55 records · Page 3Linked to original sources

PubMed1995

Immunoelectron-microscopic study of G-protein distribution in photoreceptor cells of the cephalopod Sepia officinalis.

Previous studies suggest that more than one single light-stimulated transduction pathway seems to be present in photoreceptor cells of invertebrates. Accordingly, more than one light-dependent G-protein has been detected in squid photoreceptor cells. Two different antibodies were used to locate the distribution of G-proteins in Sepia photoreceptors. One antiserum (anti-G alpha-common) has been raised against the peptide CGAGESGKSTIVKQMK. This sequence is found in most G alpha-protein subunits, it is also present in transducin of vertebrates. This sequence however, is conserved only partially in G alpha q from photosensory membranes of the squid. The second antiserum (anti-G alpha q) has been raised against the peptide QLNLKEYNLV. This sequence is present at the C-terminus of e.g. G alpha q from mouse brain and is identical also in squid. Anti-G alpha q very strongly interacted with the rhabdomeres of the photoreceptor cells from Sepia. In the non-rhabdomeric cytoplasm of the photoreceptor cells gold granules (bound to the second antibody) were almost absent. With anti-G alpha-common, only a small number of gold particles could be found at the rhabdomeres. The non-rhabdomeric cytoplasm was not labelled. Thus, further G-proteins that might be involved in a light dependent phototransduction mechanism besides the phosphoinositide pathway can hardly be expected to belong to the group of 'common' G-proteins.

Amino Acid Sequence↗

PubMed1995

Transport of endocytosed material into melanin granules in cultured choroidal melanocytes of cattle--new insights into the relationship of melanosomes with lysosomes.

Cultured choroidal melanocytes from cattle where incubated with gold labeled albumin. After phagocytosis of the labeled protein, the label appeared inside the melanin granules, as was observed under the electron microscope. Melanin granules associated with gold particles were also exocytosed into the culture medium by the melanocytes. The results of this study show that endosomes or phagosomes are transported from the cell surface of a melanocyte to the melanin granule. Therefore, melanin granules are part of the lysosomal degradation pathway. The possibility that albumin is degraded by proteases present in lysosomes and melanosomes and that the tyrosine released during degradation is used as a substrate by tyrosinase and thereby converted to melanin is discussed. The present study additionally shows that the choroidea of cattle can be used as a source for cell culture of melanocytes.

Animals↗

PubMed1994

On the Ca(2+)-dependence of inositol-phospholipid-specific phospholipase C of microvillar photoreceptors from Sepia officinalis.

The breakdown of phosphatidylinositol 4,5-bisphosphate (PIP2) by the action of a phosphoinositide-specific phospholipase C is a key event in the signal transduction mechanism of microvillar photoreceptors. The enzymatic activity of phospholipase C from photoreceptor cell membranes of the cuttlefish Sepia officinalis was measured at different concentrations of free calcium. A bell-shaped curve with a maximum of enzyme activity at about 0.2 to 0.7 microM of free calcium was obtained. In the range of 10 to 90 microM free Ca2+, the activity of the phospholipase C decreased to about 30%. The activity of the phospholipase was also modulated by strontium ions. The presence of 10 microM Sr2+ strongly reduced the dependence of the phospholipase from a given calcium concentration. Similarly to 10 mM EGTA, the presence of 10 microM La3+ reduced the enzymatic activity more than ten-fold. A comparison of these values with the changes of the calcium concentration measured in ventral photoreceptor cells of Limulus polyphemus in the dark (approximately 0.5 microM Ca2+) and under illumination (up to 30-100 microM Ca2+) indicates that in microvillar photoreceptor cells an elevation of the calcium concentration in the physiological range reduces the activity of the phospholipase C. The IP3 produced by the action of phospholipase C is known to elevate the intracellular Ca2+ concentration transiently, which in turn will be able to reduce the further IP3 formation. A negative feedback in this early part of the enzyme cascade of vision in invertebrates may substantially contribute to the process of adaptation.

Animals↗

PubMed1994

A newly discovered pathway of melanin formation in cultured retinal pigment epithelium of cattle.

According to a recent hypothesis the melanin granules in the retinal pigment epithelium of mammals originate from photosensory membrane degradation. To test this hypothesis the retinal pigment epithelium of cattle was kept in tissue culture and exposed to gold-labelled rod outer segments. Gold granules were later detected inside phagosomes, melanosomes and mature melanin granules. Tyrosinase, the key enzyme in melanogenesis, was additionally localized inside phagosomes. These results indicate that in cultured retinal pigment epithelium the matrix of the melanosome can originate from phagosomes. Therefore, the melanosome is a specialized lysosome.

Animals↗

PubMed1994

Fine structure of melanogenesis in the ink sac of Sepia officinalis.

The ink sac epithelium of the cuttlefish Sepia officinalis was investigated by electron microscopy. Melanogenesis in a simplified view seems to follow the general scheme of melanin formation in vertebrates. First, a membrane-bound protein matrix is formed, which is called an early stage melanosome. The early stage melanosomes are more or less irregular in shape with a size up to 1.5 microns and contain membranous, granular, or vesicular material. They seem to originate from Golgi bodies and/or endoplasmic reticulum. Membranes that frequently are present in the early stage melanosomes may originate from fusion of vesicles or from incorporation of Golgi membranes into early stage melanosomes. Free cytoplasmic material or mitochondria probably are also incorporated into the early stage melanosomes or melanosomes. Therefore, the origin of the early stage melanosomes seems to be similar to that of autophagosomes. The early stage melanosomes mature to melanosomes in which several dozen melanin granules are formed. These melanosomes, at last, release the melanin granules together with other cellular material, including early stage melanosomes, into the lumen of the ink gland. This finding confirms the earlier postulated holocrine character of the release. Active tyrosinase was localized in the lumen of the ink sac as already shown by biochemical methods. There was also additional evidence that most of the material of broken down cells inside the lumen of the ink sac seems to be converted into melanin granules.

Animals↗

PubMed1994

Transformation of albumin into melanin by hydroxyl radicals.

Bovine serum albumin (BSA) was exposed to hydroxyl radicals generated by the Fenton reaction. The reaction of the BSA solution with hydroxyl radicals resulted in a colour change from clear transparent to dark brown. The reaction was followed spectrophometrically. It was observed that during the reaction of BSA with hydroxyl radicals, a melanin-like absorption spectrum developed. The reaction with ferricyanide and the dark brown BSA solution resulted in the same dark blue-green colour as is typical for melanin. The results suggest that, by the reaction of BSA with hydroxyl radicals, melanin was formed.

Electrophoresis, Polyacrylamide Gel↗

PubMed1993

Localization of subrhabdomeric haemolymph lacunae in the retina of Drosophila melanogaster and Calliphora erythrocephala.

The haemolymph of flies and other insects contains tyrosinase (EC 1.14.18.1), the rate-limiting enzyme in mammalian melanogenesis. After incubation with 5 mM L-DOPA for several hours the endogenous tyrosinase of the haemolymph forms an electron dense reaction product. This method was used to localize spaces in the retina of the wild type and the white (w) mutant of Drosophila melanogaster that are filled with haemolymph. A network of subrhabdomeric haemolymph lacunae was found. Moreover it was found that these haemolymph lacunae also form extensions into the photoreceptor cells and are connected with small haemolymph lacunae that cross the retinal basement membrane. In a second set of experiments L-DOPA was injected into the thoraces of Calliphora erythrocephala. Half-an-hour after injections the flies were killed and the eyes were embedded for electron microscopy. The small molecule of DOPA or its product dopachrome, surprisingly, penetrated the retinal basement membrane and reached the subrhabdomeric haemolymph lacunae and the ommatidial cavity.

Animals↗

PubMed1993

Cyclic 3',5'-nucleotide phosphodiesterase: cytochemical localization in photoreceptor cells of the fly Calliphora erythrocephala.

The distribution of cyclic 3',5'-nucleotide phosphodiesterase activity was determined in photoreceptor cells of the fly Calliphora erythrocephala. With cAMP as substrate, staining was most intense within the phototransducing region of these cells, the rhabdomeral microvilli and also in the extracellular space surrounding the microvilli and in the mitochondria. With cGMP as substrate, the intensity within the rhabdomeres was less marked, while their extracellular surroundings were stained heavily. Thus, compared to cGMP, cAMP is the better substrate for the phosphodiesterase in the rhabdomeres of the fly. For comparison, the same cytochemical method was used to localize the well-known phosphodiesterase activity in retinal tissue of the mouse. Under the same conditions as used for fly photoreceptors, a very intense reaction product was obtained in rod outer segments. With regard to the conflicting reports concerning the light-stimulated changes of cyclic nucleotides in invertebrate photoreceptor cells, the results presented here further argue for an important role of a cyclic nucleotide in the process of phototransduction of invertebrates.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1993

Atypical granules in the eyes of the white mutant of Drosophila melanogaster are lysosome-related organelles.

In the pigment cells of the white mutant of Drosophila melanogaster, as described earlier, two types of abnormal granules are found by conventional electron microscopy. However, both types of abnormal granules, in addition to those in pigment cell invaginations, are also present in the cytoplasm of the photoreceptor cells. Three enzymes (acid phosphatase, peroxidase, and tyrosinase) are localized within the eyes of wild type and white mutant Drosophila melanogaster by electron microscopy. Peroxidase activity is present in lamellar bodies close to the rhabdomeral microvilli of both fly types. However the organelles containing peroxidase activity are 6-fold more frequent in the wild type than in the mutant. Acid phosphatase is present in lamellar bodies between and at the bases of the rhabdomeral microvilli of the wild type, as well as in ommochrome granules of the photoreceptor cells. In the white mutant, however, acid phosphatase was located in electron lucent vacuoles in the cytoplasm of the receptor cells. These acid phosphatase-positive vacuoles also contained both types of abnormal granules. The latter result indicates that abnormal granules in the receptor cells originate from lysosomal degradation and that targeting of lysosomal enzymes is altered in the white mutant. Due to the tyrosinase activity in the hemolymph of flies, the extracellular spaces are electron dense after DOPA incubation. Since some abnormal granules within the photoreceptor cells are not surrounded by an extracellular space, they can be assumed to originate within the photoreceptor cells.

Acid Phosphatase↗

PubMed1993

Does melanin turnover occur in the eyes of adult vertebrates?

This paper is a review of what is known about the turnover of melanin in iris, choroid, and retinal pigment epithelium (RPE) of the adult vertebrate eye. Differences in size and structure of choroideal and retinal pigment epithelial melanin granules are shown by electron micrographs. The classical stages of melanin synthesis, including the premelanosome, are shown in the RPE of adult hamsters that had been exposed to intense light. Degradation or synthesis of melanin also seem to occur in the melanocytes of the choroid in these animals. It is postulated that all three pigmented eye tissues (iris, RPE, and choroid) of adult vertebrates form melanin granules in vivo. However, nothing is known about the amount of this turnover.

Aging↗

PubMed1992

Isolation of the rhabdomeral microvillar cytoskeleton of the crayfish (Orconectes limosus) photoreceptor by a crosslinking reagent.

1. Rhabdomeral microvilli of photoreceptor cells of invertebrates contain a labile central cytoskeleton. For stabilization of the rhabdomeral cytoskeleton of the crayfish Orconectes limosus the crosslinking reagent suberic acid bis (N-hydroxysuccinimide ester) was used. 2. It was found that this crosslinking reagent can be successfully used to stabilize and isolate the microvillar cytoskeleton of crayfish photoreceptors. 3. After detergent treatment cytosolic proteins and the cell membranes were removed. 4. By the combined use of crosslinker and detergent the accessibility of antibodies or other markers to the microvillar cytoskeleton is possible. 5. This method may be useful, because at present little is known about the proteins associated with the central filament of invertebrate photoreceptors.

Animals↗

PubMed1992

Evidence for melanogenesis in the retinal pigment epithelium of adult cattle and golden hamster.

1. The ultrastructure of the retinal pigment epithelium (RPE) of adult Syrian golden hamsters and cattle was examined with respect to pigment granules and phagosomes involved in degradation of disk membranes from rod outer segments. 2. In the RPE of cattle, phagosomes were found that contained an electron-dense melanin-like material that was not autofluorescent and therefore not lipofuscin. 3. Disk membranes of rods are about 4 nm thick and become enlarged (7-20 nm) and electron-dense during degradation in the RPE. 4. Additionally electron-dense vesiculo-globular bodies (10-100 nm) were found in phagosomes during disk membrane degradation and in mature melanin granules. 5. In the RPE of adult hamsters that had been exposed to intense light, premelanosomes containing unmelanised filaments with a striated periodicity were found in the cytoplasm or in association with mature melanin granules. Early and late stage melanosomes were also present. Phagosomes in the RPE contained degraded disk membranes, melanin-like material and melanofilaments. 6. Dopa oxidase was detected ultrastructurally within shed disk membranes that were in close contact with the microvilli of the RPE. 7. The possibility of melanogenesis within phagosomes during disk membrane degradation is discussed.

Animals↗

PubMed1992

Localization of peroxidase activity in the retina and the retinal pigment epithelium of the Syrian golden hamster (Mesocricetus auratus).

1. Endogenous peroxidase was localized by electron microscopy within the retinal tissue of the golden Syrian hamster. 2. Peroxidase activity was found in the apical parts of rod outer segments, in the cytoplasm and phagosomes of the retinal pigment epithelium (RPE), and extracellularly between the basal infoldings of the RPE. 3. No reaction product was detected when either H2O2 or tetramethyl benzidine were omitted from the reaction medium. 4. The same was true when the tissue was osmicated before the incubation in the reaction medium or treated with 100 mM sodium azide. 5. These results indicate that the peroxidatic activity was enzymatically mediated.

Animals↗

PubMed1992

Diabetes manifestation in BB rats is preceded by pan-pancreatic presence of activated inflammatory macrophages.

Pancreata of normoglycemic diabetes-prone (dp) and diabetes-resistant (dr) BB rats and normal Wistar rats were screened for macrophage infiltration by immunohistochemistry and by electron microscopy. Inflammatory macrophages were found in the endocrine as well as the exocrine part of the pancreata from dp BB rats. In the exocrine tissue they had a different phenotype (ED1+, ED2+, W3/25+, Ox17+) from that found in pancreata from dr BB and Wistar rats (predominantly ED1-, ED2+, W3/25+, Ox17-). The number of macrophages in exocrine portions of pancreata from the various rat strains were not different. By electron microscopy scattered macrophage-associated tissue lesions and phagocytosis of cell debris were found throughout the exocrine tissue and in islets of dp BB rats. Such lesions were low or absent in biopsies of animals that later did not develop diabetes. We conclude that macrophage-mediated cytotoxicity during the early phases of diabetes development in BB rats is not restricted to islets, but is a generalized, pan-pancreatic event.

Animals↗

PubMed1991

Peroxidase and tyrosinase are present in secondary lysosomes that degrade photosensory membranes of the crayfish photoreceptor: possible role in pigment granule formation.

Three enzymes (acid phosphatase, peroxidase, and tyrosinase) were localized by electron microscopy within the retina of crayfish Orconectes limosus. Peroxidase activity was observed only in lamellar bodies, which are secondary lysosomes and degrade photosensory membrane. After H2O2 was omitted from the reaction medium, peroxidase activity in lamellar bodies was partly inhibited but was not missing completely. After addition of sodium pyruvate, which inhibits endogenous generation of H2O2, staining of lamellar bodies was absent. Tyrosinase activity was found in lamellar bodies and in small vesicles within the rhabdoms similar to those found positive for acid phosphatase. Granules (500-700 nm in diameter) with an electron opaque matrix and mature screening pigment granules showed tyrosinase activity. Moreover, lamellar structures within membrane-bound organelles that additionally contained screening pigment-like granules were electron dense because of tyrosinase activity. After addition of phenylthiourea (PTU) to the incubation medium, lamellar bodies did not generally contain electron dense deposits, although weak staining of single membranes still was sometimes observed. After addition of sodium pyruvate in combination with PTU, no staining was detected. The possible role of tyrosinase in ommochrome synthesis within secondary lysosomes that degrade photosensory membrane is discussed.

Acid Phosphatase↗

PubMed1990

Essential contribution of macrophages to islet cell destruction in vivo and in vitro.

A number of observations indicate an essential role of macrophage activity in the development of hyperglycemia in animal models of Type I diabetes. Administration of macrophage-toxic silica particles prevents spontaneous diabetes development in BB rats or NOD mice. The same result was noted in the low-dose streptozotocin-induced diabetes model in mice. Macrophages appear to be the first immune cells infiltrating islets during early insulitis. Macrophages in inflamed islets of BB rats bear the ED1 marker, whereas resident islet macrophages are ED2-positive. In vitro, ED1-positive macrophages were found to lyse pancreatic islet cells to a similar degree to various tumor cells but not normal thyrocytes. Macrophage-mediated lysis of islet cells was inhibited in the presence of 10-100 mM nicotinamide.

Animals↗

PubMed1990

Further evidence for synthesis of screening pigment granules involved in the photosensory membrane turnover of the crayfish photoreceptor.

Photosensory membrane degradation in crayfish occurs at first in multi-vesicular bodies (MVBs) and then, with the aid of lysosomal enzymes, in lysosome related lamellar bodies. In organ culture experiments with the isolated crayfish retina (Orconectes limosus) small screening pigment-like granules became visible under the electron microscope in such lamellar bodies and suggested a possible relation of photosensory membrane degradation and screening pigment granule synthesis. Chloroquine, an inhibitor of lysosomal activity, when added to the culture medium reduced the appearance of screening pigment-like granules in lamellar bodies, but led to the appearance of these granules in mature MVB's, indicating the involvement of lysosomal enzymes in the formation of pigmented lamellar bodies. In a second set of experiments the effect of bright light on the screening pigment granule ultrastructure of crayfish phoreceptors was investigated. It was found that after bright light exposure large numbers of little screening pigment granules (0.15-0.3 microns) were located between or close to rhabdomeral microvilli that were not at these sites in crayfish kept under natural light. MVB's were also reduced in size, and among the little screening pigmentary organelles granules of different electron density and morphology appeared. Additionally, vesicle flux to little screening pigment granules was detected. The screening pigment granules of the little type did not seem to be transported close to or between the microvilli, but appeared to be synthesized at these sites within little MVBs.

Animals↗

PubMed1987

Lectin binding studies on adult filariae, intrauterine developing stages and microfilariae of Brugia malayi and Litomosoides carinii.

Sections of macrofilariae of Brugia malayi and Litomosoides carinii revealed binding of the gold-labelled lectins WGA, DBA and PNA. Specificity of binding was controlled by competitive inhibition with the respective sugars. N-acetyl-glucosamine, N-acetylgalactosamine and galactose residues seem to be present in the respective tissues. The lectins were bound preferentially to parts of the reproductive organs and to the fluid contents of their lumina. The results of the chitosan test and binding experiments with WGA-gold conjugate suggest the presence of chitin in the sheath of oocytes or zygotes. Binding of WGA could not be inhibited with 0.5 M N-acetylglucosamine, but only with 10 mM triacetyl chitotriose. In older stages, binding of WGA to the sheath could be inhibited by 0.5 M N-acetylglucosamine. In mature microfilariae, the outer surface of the sheath did not show affinity for WGA, but small amounts were bound to the inner surface. Therefore, the sheath of later developmental stages and microfilariae does not contain chitin but only N-acetylglucosamine residues. The degradation of the chitin content might enable the elongation and flexibility of the sheath of microfilariae.

Animals↗