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C T Lin

Publications and source records attributed to C T Lin.

At least 181 records · Page 10Linked to original sources
PubMed1985

Is taurine a neurotransmitter in rabbit retina?

Rabbit retina was used as a model to study the possible role of taurine in the retina. The taurine-synthesizing enzyme, cysteine sulfinic acid decarboxylase (CSAD), is localized immunohistochemically using specific antibodies against CSAD. The CSAD-immunoreactivity appears to be most prominent in the inner nuclear layer (INL) and ganglion cell layer (GCL). The inner plexiform layer (IPL), the outer nuclear layer and outer plexiform layer are sporadically stained. The CSAD-positive neurons include some amacrine cells and probably the bipolar cells in the INL and some large and small ganglion cells in the GCL. Autoradiographic studies reveal that the uptake of [3H]taurine is most prominent in the INL. The IPL and GCL, as well as the Müller cells, also show a moderate degree of [3H]taurine accumulation. In conclusion, we have demonstrated the presence of the taurine-synthesizing enzyme and uptake systems in rabbit retina. Based on the above evidence, we propose that taurine may be used by some neurons, presumably amacrine cells, as a transmitter in the rabbit retina.

Animals↗
PubMed1985

Ultrastructural demonstration of L-glutamate decarboxylase and cysteinesulfinic acid decarboxylase in rat retina by immunocytochemistry.

The gamma-aminobutyric acid (GABA) synthesizing enzyme, L-glutamate decarboxylase (GAD), and the taurine synthesizing enzyme, cysteinesulfinic acid decarboxylase (CSAD) have been localized in rat retina at the ultrastructural level by indirect immunoelectron microscopy. GAD immunoreactivity (GAD-IR) was seen only in some amacrine cells and their terminals. CSAD immunoreactivity (CSAD-IR) was found in most retinal neuronal types and their processes including photoreceptor cells (rod and cone cells), bipolar cells, amacrine cells and ganglion cells. The GAD-IR positive amacrine terminals have been found to make synaptic contact with other GAD-IR negative bipolar and amacrine terminals, and ganglion cell dendrites. Most of the GAD-IR positive terminals are presynaptic. Occasionally, GAD-IR positive amacrine terminals are postsynaptic to another amacrine terminal or ganglion cell body. In the inner plexiform layer, CSAD-IR positive amacrine terminals also make synaptic contacts with other nerve terminals, similar to that of GAD-IR positive amacrine terminals. In addition, CSAD-IR positive bipolar terminals make synaptic contact with some CSAD-IR positive as well as negative amacrine terminals. Both CSAD-IR positive amacrine and bipolar terminals are mostly presynaptic to other CSAD-IR negative terminals. In the outer plexiform layer, CSAD-IR was found to be associated with synaptic vesicles and the synaptic membrane in certain cone pedicles and rod spherules. It is concluded that only a fraction of amacrine cells in rat retina may use GABA as a neurotransmitter. The presence of CSAD-IR in some amacrine, bipolar, photoreceptor and ganglion cells in rat retina is compatible with the notion that taurine may play some important roles, such as those of neurotransmitter or neuromodulator in mammalian retina.

Animals↗
PubMed1985

Immunoelectron microscopic demonstration of prostatic acid phosphatase in human hyperplastic prostate.

Immunoelectron microscopic studies were done on prostatic tissues obtained from patients with benign hyperplasia. Rabbit IgG-peroxidase conjugate against purified human prostatic acid phosphatase band 2 (HPAP-2) was used for studies. Under the light microscope, the columnar secretory epithelia of prostatic glands showed different intensity and distribution of immunostaining whereas the basal cells were unstained. Under the electron microscope, the secretory epithelial cells often showed electron-dense reaction product in the Golgi apparatus and secretory vesicles and vacuoles, and only sparingly in the cisternae of nuclear envelope and rough ER. Sometimes, fusion of secretory vacuolar membrane and plasma membrane and discharge of the vacuolar contents into the extracellular space were noted. The surfaces of microvilli at the apical portion of the columnar epithelia and the lumen of the glandular acini always showed reaction product. These findings suggest that HPAP-2 may be synthesized in the rough ER and transported to the Golgi apparatus where it is concentrated and transferred to the secretory vesicles and vacuoles. HPAP-2 is finally discharged into the extracellular spaces through exocytosis, a secretory mechanism similar to that of other secretory proteins.

Acid Phosphatase↗
PubMed1983

Immunocytochemical localization of L-glutamate decarboxylase, gamma-aminobutyric acid transaminase, cysteine sulfinic acid decarboxylase, aspartate aminotransferase and somatostatin in rat retina.

The regional distribution and cellular location of GABA-synthesizing enzyme, L-glutamate decarboxylase (GAD), GABA degrading enzyme, GABA-transaminase (GABA-T), taurine synthesizing enzyme, cysteine sulfinic acid decarboxylase (CSAD), aspartate and glutamate converting enzyme, aspartate aminotransferase (AAT), and somatostatin have been visualized in the rat retina by immunocytochemical methods. GAD immunoreactivity was found to be concentrated in the inner plexiform layer. A moderate to weak staining of GAD was found in the inner nuclear layer. The distribution of GABA-T immunoreactivity was similar to that of GAD with the exception that a weak to moderate staining of GABA-T was also observed in the outer plexiform layer. CSAD immunoreactivity was seen in every layer with the heaviest staining in the inner plexiform layer, and moderate staining in the inner and outer nuclear layers and ganglion cell layer. AAT immunoreactivity was mostly concentrated in the outer nuclear layer; there was weak staining in the inner nuclear layer and inner and outer plexiform layer. Dense somatostatin staining was seen in the inner plexiform layer and moderate staining was present in the inner nuclear layer, outer plexiform layer and ganglion cell layer. These findings suggest that in rat retina, GABA-containing cells occur in some types of amacrine cells only, while taurine and somatostatin appear in both amacrine and horizontal cells. AAT immunoreactivity was primarily associated with the photoreceptor cells suggesting that AAT may be used as a marker for aspartergic/glutamergic cells and their endings in the central nervous system.

4-Aminobutyrate Transaminase↗
PubMed1983

Immunoelectron microscopic study of somatostatin biosynthesis in dog pancreas.

The biosynthesis of somatostatin has been studied at the ultrastructural level in pancreatic islets by using rabbit antiserum against synthetic somatostatin. To document that the antiserum specifically bound preprosomatostatin, we have tested the ability of the antiserum to precipitate the product synthesized in vitro. Poly(A) enriched RNA isolated from catfish islets was translated in both the wheat germ extract and nuclease-treated reticulocyte lysate systems. It was found that the in vitro translation product, preprosomatostatin, could be recognized by the antibody against synthetic somatostatin. The morphological study was then performed by immunoelectron microscopy by using the Fab-peroxidase conjugate technique. In dog pancreatic islets, somatostatin immunoreactive reaction product was seen only in the delta cells. In these cells, they were detected on bound ribosomes, in the cisternae of the rough endoplasmic reticulum (ER) and Golgi apparatus, in the Golgi associated vesicles, and in secretory vesicles. These findings suggest that somatostatin precursor molecules are synthesized on bound ribosomes and discharged into the cisternae of the rough ER. They are then transported to the Golgi apparatus and transferred to the secretory vesicles for secretion. The different staining intensities in the secretory vesicles would suggest that the processing of the precursor molecules of somatostatin probably takes place in the secretory vesicles.

Animals↗
PubMed1983

A comparative study of polyclonal and monoclonal antibodies for immunocytochemical localization of cytosolic aspartate aminotransferase in rat liver.

The rabbit antiserum and mouse monoclonal hybridoma antibody against porcine cytosolic aspartate aminotransferase (c-AAT) (or cytosolic glutamic oxaloacetic transaminase (c-GOT)) were produced and compared for the localization of c-AAT in rat liver. An indirect immunocytochemical technique was performed using peroxidase-conjugated goat immunoglobulin (Ig) G anti-rabbit IgG and peroxidase-conjugated rabbit IgG anti-mouse IgG as the second antibody. Rats were perfused with paraformaldehyde-lysine-periodate fixative and the liver fragments were immersed in 4% paraformaldehyde and transferred to 10% dimethyl sulfoxide overnight and subjected to cryostat sectioning. The rabbit IgG antibody, 3 individual monoclonal antibodies, and a mixture of these 3 monoclonal antibodies were applied to the tissue sections, respectively, using the same concentration. Under the same experimental conditions, the c-AAT was localized in each individual hepatocyte by both monoclonal and polyclonal antibodies. However, a mixture of three monoclonal antibodies gave stronger staining than a single monoclonal antibody; although two antibodies yield more intense staining than just one, it was still less intense than for three. The conventional rabbit polyclonal antibody against c-AAT produced more reaction product than the combined three monoclonal antibodies. It is concluded that for immunocytochemical study, the use of a single monoclonal antibody is sensitive enough to localize its tissue antigen under the present experimental condition. To obtain a stronger reaction product, a combination of several monoclonal antibodies, at least three or more, may give better staining.

Animals↗
PubMed1983

Production and characterization of an antibody to cytosolic aspartate aminotransferase and immunolocalization of the enzyme in rat organs.

Cytosolic aspartate aminotransferase (c-AAT) was purified to homogeneity from porcine heart and immunized to rabbit for production of antiserum. The purity of this enzyme protein and the specificity of its antibody were judged by silver-stained-sodium dodecyl sulfate slab gel, Western blot transfer technique, and double immunodiffusion. The antibody against porcine heart c-AAT was found to cross-react with rat c-AAT but not with nine other different enzymes from the heart, liver, and muscle. Affinity purified antibody was used to localize this isoenzyme in the rat heart, liver, kidney, and cerebellum by indirect immunoperoxidase method. It was found that, in the rat heart muscle, c-AAT reaction product was present as a linear structure parallel to the muscle fiber and along the sarcolemma. Some cardiac muscle fibers contain more reaction products than the others. In the liver, reaction product was seen unevenly distributed in the hepatocytes. The Kupffer cells and endothelia were less stained. Most of the tubular epithelia of the loop of Henle in the kidney were intensely stained. But other tubular epithelia including convoluted and collecting tubules were sporadically and less stained. The basket and stellate cells and their neuronal processes and terminals in the cerebellum were markedly stained, but the Purkinje and granule cell bodies were weakly stained. For comparison of the staining intensity with enzyme activity in each organ, the c-AAT enzyme activity was simultaneously determined in those organs. This study indicates that the presence of c-AAT is specific in different organs and tissues.

Animals↗
PubMed1982

Studies of hCG binding and endocytosis in rat ovary by ultrastructural immunocytochemistry.

Localization of hCG binding sites and the process of endocytosis in pseudopregnant rat ovaries were investigated by indirect electron-microscopic immunocytochemistry. Immature female rats were treated with pregnant-mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) to induce ovarian luteinization. Eight days after priming with PMSG-hCG and 1-6 h before sacrifice the animals were given another injection of hCG to bind the receptors. Receptor sites to hCG localized by reaction product were present in most luteal cells, but not in primary follicular cells. The receptor sites were distributed on luteal cell surfaces facing interstitial spaces. Endocytotic pits containing hCG binding sites were rarely seen 1 h after hCG injection. At 2h, hCG and presumably its receptor were taken up within endocytotic vesicles with the evidence of reaction product coated on the vesicle wall. With time, fusion of endocytotic vesicles with lysosome occurred and the reaction product appeared in phagolysosomes. The reaction product was localized on phagolysosomal inner surface or in free granular form. These findings suggest that hCG and its receptors were internalized through endocytotic pits and endocytotic vesicles and delivered to lysosomes probably for degradation. An additional experiment for localization of acid phosphatase was also performed to delineate the lysosomes and phagolysosomes.

Animals↗
PubMed1982

Localization of apoVLDL-II, a major apoprotein in very low density lipoproteins, in the estrogen-treated cockerel liver by immunoelectron microscopy.

We have studied the sites of synthesis, assembly, and secretion of apoVLDL-II, a major apoprotein in very low density lipoproteins (VLDL), in the cockerel liver by immunoelectron microscopy. In the liver of the estrogen-treated cockerel, apoVLDL-II reaction products were localized in the cisternae of the nuclear envelope and the rough endoplasmic reticulum (RER). Such products were not observed in the smooth endoplasmic reticulum (SER). ApoVLDL-II reaction products were also located on the surface of lipid particles in the Golgi apparatus and secretory vesicles. Such lipid particles were not detected in the RER or SER. Some secretory vesicles containing the reaction products were seen during the process of fusion with the plasma membrane. Such fusion took place against the plasma membrane lining the space of Disse as well as the intercellular spaces. Reaction products also occurred in the sinusoids. These observations are compatible with the following sequence of events in the synthesis, assembly and secretion of apoproteins in VLDL in the cockerel liver: ApoVLDL-II is synthesized on bound ribosomes attached to the nuclear envelope and RER, and is discharged into their cisternae. The protein is probably transported to the Golgi apparatus where the assembly of this protein and its lipid components probably takes place. Secretory vesicles derived from the Golgi apparatus carry the VLDL particles to the plasma membrane where secretion of these particles takes place by exocytosis, and the VLDL are discharged into the sinusoid via both the space of Disse and intercellular spaces.

Animals↗
PubMed1982

Taurine in the mammalian cerebellum: demonstration by autoradiography with [3H]taurine and immunocytochemistry with antibodies against the taurine-synthesizing enzyme, cysteine-sulfinic acid decarboxylase.

Taurine neurons and their dendrites and axons were visualized in the mammalian cerebellum by autoradiography, after in vivo injections of [(3)H]taurine directly into the cerebellar cortex or deep cerebellar nuclei, and by immunocytochemistry at the light- and electron-microscope levels with antibodies against cysteine-sulfinic acid decarboxylase (CSADCase; L-cysteine-sulfinate carboxylyase, EC 4.1.1.29). Uptake and sequestration of [(3)H]taurine labeled numerous Purkinje cell somata, primary dendrites, and axons; many granule cell somata, dendrites, and parallel fibers; stellate, basket, and Golgi cells; the larger neurons in all deep cerebellar nuclei; the largest neurons in the lateral vestibular nucleus; and, more rarely, Purkinje cell axonal terminals in the neuropil. The label at all sites was diminished by preinjection into the cerebellum of hypotaurine, p-chloromercuriphenylsulfonic acid, or beta-alanine, and was virtually eliminated by strychnine. Immunocytochemical labeling with polyclonal antibodies directed against CSADCase, the enzyme responsible for the synthesis of hypotaurine from cysteine sulfinic acid and taurine from cysteic acid, had a similar distribution. In electron micrographs, immunoreactivity within Purkinje cell somata and dendrites was localized to the Golgi apparatus, the inner plasma membrane, and condensed nonmembranous foci (120 nm in diameter) marked by clumps of peroxidase reaction product. Large Nissl bodies were usually not CSADCase immunoreactive. Numerous immunoreactive granule cells, dendrites, and parallel fibers were recognized. Pretreatment of the animals with colchicine increased the intensity of CSADCase immunoreactivity but did not change the number or distribution of labeled cells. These experiments indicate that taurine is synthesized and involved in a specific uptake process by cerebellar neurons. Neuroglial cells do not synthesize taurine but some neuroglia take up [(3)H]taurine. These findings call for a reexamination of the physiological function of taurine in the cerebellum. A hypothesis is proposed that taurine may be involved in the regulation of calcium, in dendritic spike generation, and in the inhibition of impulse propagation in major Purkinje cell dendrites.

Animals↗
PubMed1982

Light and electron microscopic immunocytochemical localization of clathrin in rat cerebellum and kidney.

The precise cellular and subcellular locations of coated vesicle protein, clathrin, in rat kidney and cerebellum have been visualized by immunocytochemical techniques. In the renal tubular epithelia, clathrin-positive products were found on both free ribosomes and on those attached to rough endoplasmic reticulum (RER) and the nuclear envelope. No clathrin was observed in the cisternae of RER or the Golgi apparatus. Clathrin-positive reaction products could also be seen on coated pits, coated vesicles, Golgi-associated vesicles, basolateral cell membrane, the ground substance, and in the autophagic vacuoles. In cerebellar Purkinje and granule cell bodies, reaction products were seen localized on coated vesicles, on the budding areas from the Golgi-associated membrane and Golgi-associated vesicles. Furthermore, the membrane of the multivesicular body, the bound-ribosomes, and the ground substance were also stained. In the myelinated axon, the clathrin appeared to be concentrated on certain segments and seemed to fill in the space between neurotubules and some vesicles. In certain synaptic terminals clathrin was often seen attached to presynaptic vesicles, presynaptic membrane, and post-synaptic membrane. However, in most mossy fibers, some synaptic vesicles were not stained. These observations suggest that clathrin is synthesized on bound and free ribosomes and discharged into the cytosol where it becomes associated with a variety of ground substances and assembles on coated pits, coated vesicles, Golgi-associated vesicles, presynaptic vesicles, and pre- and postsynaptic membranes. Clathrin may be finally degraded in autophagic vacuoles.

Animals↗
PubMed1982

Ultrastructural and lysosomal enzyme studies of skeletal muscle and myocardium in rats with long-term vitamin E deficiency.

Muscular dystrophy and cardiomyopathy were produced in weanling rats by feeding a vitamin E-deficient diet for 12 mth. Deficient and control rats were killed, and skeletal muscle and myocardium were used for subcellular studies and biochemical assay of selected lysosomal enzymes. Ultrastructurally, the skeletal muscle showed various degrees of pathological changes. In the severely damaged muscle fibres, prominent increase of secondary lysosomes, autophagic vacuoles, residual bodies, disappearance of myofilaments, rupture of sarcolemma and shrinkage of muscle fibres were noted. The damaged muscle fibres finally became dense residual bodies and dispersed in the interstitial spaces, where the macrophages and fibroblasts were found. In the myocardium, some muscle fibres were intact with mild fatty infiltration and marked proliferation of mitochondria. However, in the severely damaged myocardial fibres, the whole fibre was always filled with amorphous dense bodies, and the sarcolemma was ruptured. This resulted in dispersion of many cellular organelles in the surrounding interstitial space. A significant increase of cathepsin and beta-glucuronidase activity in the cytosol of both organs suggests that lysosomal enzymes may play a major role in the destruction of muscle and cardiac fibres in the long-term vitamin E-deficient animals.

Animals↗
PubMed1981

Estrogen regulation of yolk and non-yolk protein synthesis in the avian liver. An immunocytochemical study.

The effects of acute and chronic estrogen treatment on two egg yolk proteins, vitellogenin and apoVLDL-II, and two non-yolk proteins, ovalbumin and apoA-I, were studied by immunocytochemical techniques. Three groups of cockerels received either no treatment, or a single injection of diethylstilbestrol (DES, 2.5 mg) (acute stimulation) 24 h before killing, or 14 daily injections of 2.5 mg DES (chronic stimulation) before killing. The animals were killed at 4 weeks of age and their livers examined with respect to the distribution of the four different proteins by the indirect immunoperoxidase method. Vitellogenin was undetectable in the untreated cockerel liver. A single injection of DES resulted in the appearance of the protein in approximately 10%-15% of the hepatocytes. Chronic DES stimulation increased the number of positive cells to about 20%. In contrast, apoVLDL-II was present in 1%-2% of the hepatocytes in untreated animals. It was detected in an increased proportion (20%-25%) of cells after a single dose of DES. After chronic estrogen treatment, there was a very marked increase in the number of positive cells (less than 90%). Ovalbumin was undetectable in untreated cockerel liver, while apoA-I was detected in an extremely low proportion of cells (0.005%-0.01%). Neither ovalbumin nor apoA-I distribution seemed to be affected by a single dose of DES. However, chronic DES treatment resulted in the appearance of ovalbumin-containing cells (approximately 0.02%) and a marked increase in the number of cells containing apoA-I (10%-15%).

Animals↗