Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In this study, we describe the distribution of actin filaments in and around spermatids that are mechanically dissociated, in the presence and absence of exogenous trypsin, from the seminiferous epithelium of the rat. NBD-phallacidin and subfragment 1 of the myosin molecule (S-1) are used as probes for filamentous actin at the light and electron microscopic levels, respectively. The fluorescence associated with spermatids mechanically dissociated in the absence of trypsin is due to actin both in the spermatogenic cells themselves and in attached Sertoli cell ectoplasmic specializations. Fluorescence generated by labelled actin in ectoplasmic specializations occurs in linear tracts that follow the outer contour of spermatid heads. The residual fluorescence seen when trypsin is used to detach Sertoli cell fragments is diffuse and due to f-actin in the subacrosomal space. Electron microscopic data agree with the fluorescence results. This study conclusively demonstrates that Sertoli cell ectoplasmic specializations remain attached to spermatids mechanically dissociated from the seminiferous epithelium. This observation may be useful when attempting to isolate ectoplasmic specializations for biochemical analyses.
Anat Rec 1987 May
PMID:Distribution of actin in spermatids and adjacent Sertoli cell regions of the rat. 360 57

Renal tubular cells and segments isolated by a trypsin, pepsin, pronase E digestion procedure were studied with scanning electron microscopy. The basal and lateral surfaces of S1, S2, S3 proximal tubular (PT) segments, descending and ascending thin limbs of Henle (TL), distal ascending thick limb of Henle, or distal straight tubule (DST) and distal convoluted tubule (DCT) segments, connecting tubules (CNT), and collecting ducts (CD) were identified and characterized. The basal processes of the S1 and S2 PT cells were fan shaped, were oriented in a circumferential direction, and terminated in microvilli at the basement membrane. S3 PT cells had microvillous basal processes mainly on the lateral edges of the cells. The basal processes of DST and DCT were similar to PT in orientation but terminated on the basement membrane with flattened, thin attachments. The long-loop descending TL and the ascending TL exhibited distinctive interdigitating cell processes. TL segments with simple contours were present in smaller numbers and were characteristic of short-loop descending limbs. CNT showed some cells with basal surfaces resembling DCT cells and others resembling CD cells. Both cortical and medullary CD segments exhibited intercalated cells with round basal contours and a sparse pattern of basal infolding clefts. The cortical CD principal cells revealed a much more elaborate mosaic of plicae, clefts, and microvilli than those of the medullary CD. These observations extend the previous knowledge gained from transmission electron microscopy and assist in the interpretation of that knowledge.
Anat Rec 1985 Oct
PMID:Scanning electron microscopy of basolateral surfaces of rat renal tubules isolated by sequential digestion. 390 17

Samples of human plantar and palmar skin were excised and incubated in 20 mM EDTA after which the epidermis was gently separated from the dermis with the plane of separation occurring in the lamina lucida. Scanning electron microscopic examination of the dermal component revealed the classically described series of regularly spaced grooves and papillae that characterize the epidermal-dermal junction in thick skin. Primary dermal grooves exhibited evenly spaced tunnels that were originally occupied by sweat gland ducts. The basement membrane (basal lamina) in the primary grooves was relatively smooth but did exhibit a flattened, reticulated pattern at high magnifications. The basement membrane of secondary dermal grooves and papillae was in the form of numerous, elevated microridges off of which septae arose at roughly right angles. The surface appearance of the basement membrane in these areas was that of a honeycomb owing to the numerous compartments and recesses formed by the ridges and septae. Degradation of the basement membrane by trypsin demonstrated that the foundation for the highly folded and compartmentalized basement membrane was composed of dermal collagen fibrils, 60-70 nm in diameter, that were arranged in a series of variably sized, interconnected collagen bundles or walls. Epidermal basal cells extended cytoplasmic (foot) processes into two or more compartments, formed by the ridges and septae, which considerably amplified the basement membrane surface available for epidermal attachment. Scanning electron microscopic studies of the epidermal-dermal junction confirm the variable surface character of this interface previously reported by others using sectioned material.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1985 Feb
PMID:Variation in basement membrane topography in human thick skin. 397 83

Studies of Sertoli cell structure, maturation, and function have been aided by the use of in vitro systems. Although numerous papers have appeared that utilize the Sertoli cell culture model, few papers have dealt with the characterization of these cells under various culture environments. Recently, it has been reported that the addition of serum to the culture medium prevents induction of long cytoplasmic appendages in cultured Sertoli cells that have been treated with FSH, TSH, or c-AMP. The purpose of this investigation was to determine which serum components, obtained by gel filtration, are capable of inhibiting the morphological response induced by FSH, TSH, or c-AMP. Sertoli cell-enriched cultures were prepared using collagenase and trypsin digestion, each followed by gravity sedimentation. Untreated cells grown on plastic or glass substrates assumed an epithelioid appearance after several days. Cells treated with FSH, TSH, or c-AMP formed long cytoplasmic appendages after 1-2 days. This response was prevented or reversed by the addition of fetal calf serum (10%), crystallized bovine serum albumin (0.25%-2%), or purified albumin obtained by gel filtration of whole serum (0.25%). It was also found that fractions that elute between the void volume and the initial albumin fractions (molecular weights of approximately 50,000 and greater) mimic the hormone-induced response after only 10-12 hours. The results of this investigation indicate that albumin is the primary serum component responsible for inhibiting morphological alterations induced by FSH, TSH, and c-AMP. Furthermore, it is apparent that the production of long filamentous cytoplasmic appendages in Sertoli cells can be induced by a wide variety of substances.
Anat Rec 1980 Jun
PMID:Effects of serum components on the morphology of Sertoli cells in culture. 625 34

The presence of structures bridging the inner and outer acrosomal membranes of the equatorial segment of boar spermatozoa was clearly demonstrated in cells that have undergone a variety of treatment procedures to displace the electron-dense contents of the acrosome. En-face sections show bridges to be punctate and not linearly extensive as might be suggested by sections perpendicular to the flat plane of the head. About 4.5 x 10(5) bridges, each measuring 7 nm across and spaced 7 nm apart, are arrayed hexagonally in the equatorial segment, but bridges are not present within the principal segment of the acrosome. Short-term treatment with trypsin partially digests the bridges, but does not disrupt the spacing or strict parallel configuration of equatorial segment membranes. However, short-term treatment with pronase digests most bridges and effectively disrupts the typical configuration of the equatorial segment. Freeze-fracture of the cytoplasmic face of the acrosomal membranes of the equatorial segment reveals a pattern throughout the phospholipid layer of the membrane which is similar to the pattern of bridges present in en-face thin sections of the equatorial segments. The data suggest that numerous bridges link the inner and outer acrosomal membranes of the equatorial segment of the acrosome and they play a major, if not an exclusive, role in maintaining the close spacing and parallel arrangement of the membranes in this portion of the acrosome.
Anat Rec 1980 Nov
PMID:On the presence of bridges linking the inner and outer acrosomal membranes of boar spermatozoa. 700 60

A double reciprocal translocation has been found in a morphologically normal but infertile bull (Bos taurus), using trypsin-Giemsa banding. The karyotype of the bull can be written as: 60,XY,t(2q-;20q+),t(8q-;27q+). Cytophotometric studies supported the findings in the karyograms and identified the breakpoints in the chromosomes involved. The possible effect of this translocation heterozygosity on male fertility is discussed.
Vet Rec 1982 Feb 27
PMID:Double reciprocal translocation heterozygosity in a bull. 707 21

When chondrocytes from the Swarm rat chondrosarcoma are isolated by trypsin and collagenase digestion and cultured in Petri dishes, they form a new extracellular matrix within 24 hours, with proteoglycan matrix granules and collagen fibrils. This rapid synthesis of new matrix, together with the biochemical and morphological characterization of the proteoglycans as typical of cartilage, demonstrates the value of these cultures as a model system for studies of synthesis, secretion, and organization of extracellular matrix.
Anat Rec 1981 Jul
PMID:The ultrastructure of cultures from the Swarm rat chondrosarcoma. 727 Sep 28

Using a collagenase trypsin-EDTA treatment, we have been able to successfully isolate and grow primary cultures of the lymphatic endothelium (LEC) that were subcultured, frozen for storage, subsequently thawed with good recovery and growth, and serially subcultured. The morphological features of cultured LEC were consistent with that observed for the endothelium of intact lymphatic vessels. A prominent feature of growing cultures was the appearance of large vacuoles in the perinuclear region of the cytoplasm, which became filled with fluid and cell debris engulfed from the culture medium. The basal cell surface lacked a well defined basal lamina and anchoring filaments were observed extending from the basal plasmalemmal surface into the underlying substratum. LEC in cultures were also positive for Factor VIII-related antigen. However, specific granules, characteristic of Weibel-Palade bodies were not observed in ultrathin sections of confluent cultures. F-actin was identified in LEC cultures using fluorescein phalloidin, and in confluent cultures actin filaments were located at the periphery of the cell as a continuous circumferential thin band and short filamentous bundles in the central part of the cell. By using heparin and endothelial cell growth supplement in the culture medium we have been able to grow stable cultures of lymphatic endothelial cells that could be maintained when serially subcultured for over two years. These LEC cultures provide an in vitro model for investigating the function and biochemical properties of the lymphatic endothelium.
Anat Rec 1993 Aug
PMID:Lymphatic endothelium isolation, characterization and long-term culture. 837 89

Cheviot sheep from the Neuropathogenesis Unit flock were examined for PrP in brain sections using immunocytochemistry in order to aid scrapie diagnosis. Brains were collected from sheep which had been naturally or experimentally infected with scrapie and fixed in periodate-lysine-paraformaldehyde or in formalin. Immunolabelling was achieved using a monoclonal antibody (FH11) raised to the N-terminus of recombinant PrP protein. Several pre-treatments were studied in an effort to enhance PrP immunolabelling such as trypsin, formic acid and hydrated autoclaving. Trypsin was successful in highlighting PrP staining in formalin-fixed tissue. PrP staining was regularly observed in the dorsal vagus nucleus of the medulla oblongata and in the thalamus. Differences in the distribution and intensity of PrP immunostaining were apparent between the scrapie sources ME7 and SSBP/I.
Vet Rec 1996 Nov 23
PMID:Immunolocalisation of the prion protein (PrP) in the brains of sheep with scrapie. 895 91

The nature of the liver binding site which is responsible for the initial recognition and clearance of chylomicron-remnants and beta-migrating very-low-density lipoprotein (beta-VLDL) is under active dispute. We have investigated the effect of the 39-kDa receptor-associated protein (RAP) on the recognition site for activated alpha 2-macroglobulin and beta-VLDL on rat liver parenchymal cells in vivo and in vitro in order to analyze whether both substrates are recognized and internalized by the same receptor system. Radiolabelled trypsin-activated alpha 2-macroglobulin (alpha 2M-T) was cleared rapidly by the liver (maximal uptake of 80.8 +/- 1.0% of the injected dose). Prior injection of 5, 15, or 50 mg gluthathione-S-transferase-linked RAP (GST-RAP)/kg rat reduced the liver uptake to 62.2 +/- 2.3%, 59.3 +/- 1.1%, or 2.9 +/- 0.1% of the injected dose, respectively. Concurrently the serum decay was strongly delayed after injection of 50 mg GST-RAP/kg rat but this did not affect the serum decay and liver uptake of 125I-beta-VLDL. Binding studies with isolated liver parenchymal cells in vitro demonstrated that the binding of 125I-alpha 2M-T was 98% inhibited by GST-RAP with an IC50 of 0.3 microgram/ml (4.2 nM), whereas the binding of 125I-beta-VLDL and 125I-beta-VLDL + recombinant apolipoprotein E (rec-apoE) was unaffected by GST-RAP up to 50 micrograms/ml (700 nM). Also, the cell association and degradation of alpha 2M-T was blocked by RAP, while the association and degradation of beta-VLDL and beta-VLDL + rec-apoE were not influenced. The inhibitory effect of RAP on the cell association and degradation of alpha 2M-T lasted for 1-2 h of incubation at 37 degrees C. The binding of the radioiodinated RAP to isolated liver parenchymal cells was highly efficiently coupled to lysosomal degradation. Upon in vivo injection into rats, 125I-labeled RAP is rapidly cleared from the serum and taken up by the liver, which is also coupled to efficient degradation. Since RAP blocks binding of all known ligands to the alpha 2-macroglobulin receptor/low-density lipoprotein receptor-related protein (the alpha 2Mr/LRP) and at high concentrations the binding to the LDL receptor, we conclude that the initial binding and internalization of beta-VLDL by rat liver parenchymal cells is not mediated by the alpha 2Mr/LRP. The properties of binding of beta-VLDL to rat liver parenchymal cells points to an apoE-specific recognition site for lipoprotein remnants which differs from the alpha 2Mr/LRP, proteoglycans and the LDL receptor and is tentatively called the lipoprotein remnant receptor.
 ...
PMID:Blockade of the alpha 2-macroglobulin receptor/low-density-lipoprotein-receptor-related protein on rat liver parenchymal cells by the 39-kDa receptor-associated protein leaves the interaction of beta-migrating very-low-density lipoprotein with the lipoprotein remnant receptor unaffected. 902


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