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Changes in electronegative and electropositive surface charges and in lectin receptors (concanavalin A and wheat germ agglutinin) were investigated on sperm plasma membranes of the monkey (Macaca fascicularis) during epididymal transit and after ejaculation. Electronegative charges at pH 1.8, which were uniformly distributed on the whole plasma membrane of caput epididymal spermatozoa, increased mainly on the postacrosomal cap and the tail during epididymal passage. Electropositive charges at pH 9 were simultaneously found on the whole cell surface of caput epididymal spermatozoa with a stronger labeling on the acrosomal apex, the postacrosomal cap, and the tail. These charges disappeared during passage through the epididymis corpus. The surface distribution of lectin receptors varied inversely during epididymal transit with an increase in concanavalin A receptors and a decrease in wheat germ agglutinin receptors. These data show that changes in the monkey sperm plasma membrane during epididymal maturation occur in the distal corpus of the epididymis.
Anat Rec 1984 Mar
PMID:A cytochemical study on surface charges and lectin-binding sites in epididymal and ejaculated spermatozoa of Macaca fascicularis. 654 30

The mouse epididymal duct can be histologically divided into five segments (I-V), and the principal cells in segment II appear to secrete periodic acid-Schiff (PAS)-positive material into the lumen. In this study, male dd-mice received one, two, or four 800-R doses of radiation beginning at age 50 days. Mice receiving multiple doses were irradiated at 1-week intervals. After irradiation, marked depletion of spermatozoa, or aspermia, occurred in the epididymal duct for 2 to 16 weeks after a latency period of 3 to 4 weeks according to the times of irradiations. During oligospermia or aspermia, PAS-positive inclusions appeared in the principal cells in segment IV. The inclusions occupied a supranuclear position and appeared as round granules and globules measuring 2-15 micron in diameter, and increased in number, size, and staining intensity with time. They disappeared after reappearance of spermatozoa. The findings suggest that PAS-positive material may bind to spermatozoa and, if not bound, is reabsorbed by the principal cells in segment IV and deposited as intracellular inclusions, and the principal cells in segment IV are capable of digesting the accumulated PAS-positive material.
Anat Rec 1983 Sep
PMID:Response of epididymal duct to the temporary depletion of spermatozoa induced by testicular irradiation in mice. 663 30

Mouse and guinea pig epididymal tissues have been investigated by light and electron microscopic autoradiography after long intervals ranging from 24 h to 5 days postinjection (p.i.) of the glycoprotein precursors, L-fucose-6-3H or D-glucosamine-1-3H. Using modified fixations to enhance glycoprotein preservation in situ, we found intense labelling of luminal contents in at least some of the epididymal segments after all the intervals investigated. At 24 h p.i., the label in guinea pig was associated with spermatozoa during remodelling of the acrosome in segment II, and at 3 days p.i., radioactivity was trapped within sperm head associations ("rouleaux") in segment IV of the epididymis. At this time, similar rouleau labelling extended from segment IV to segment VIII. In mouse, the luminal contents of the cauda epididymis were still intensely labelled at 5 days p.i.; analysis of the electron microscopic autoradiograms showed that relative grain concentration over the spermatozoa was twice that of the epididymal plasma. This concentration was especially elevated in the region of the sperm head. These findings taken together were interpreted as the binding of secreted epididymal glycoproteins to spermatozoa during sperm transit through the epididymis. In contrast to luminal contents, the labelling of the epididymal epithelium was generally lower, except on the clear cells which showed more pronounced labelling than the neighboring principal cells in mouse cauda epididymis at 5 days p.i. This label probably originated from the resorption of luminal glycoproteins.
Anat Rec 1984 Feb
PMID:Binding of secreted glycoproteins to spermatozoa in the mammalian epididymis: a fine-structure autoradiographic study. 670 37

Antisera against Actinobacillus seminis, Brucella ovis and Corynebacterium pseudotuberculosis were prepared in adult female goats. Specific immunofluorescence was observed in cultural smears of A seminis, B ovis and C pseudotuberculosis by the direct technique and in smears of A seminis also by the indirect technique. Individual organisms could be recognised. Specific fluorescence of A seminis was readily detected in semen. The results indicate that immunofluorescence may offer an effective method for rapidly and accurately diagnosing bacterial epididymitis in sheep, especially before epididymal lesions are palpable.
Vet Rec 1980 Nov 01
PMID:Diagnosing ovine epididymitis by immunofluorescence. 702 Feb 28

An improved method is presented for processing single cells for electron microscopy. Agarose, which has a low (30 degrees C) gelling temperature, was used as an initial embedding medium for single cells (spermatozoa and oocytes) and dissociated cell preparations (luteal cells and spleen cells). Dispersed cells of corpus luteum, spleen, and epididymal spermatozoa were placed in 1.5% agarose after aldehyde fixation. These fixed cells, embedded in agarose, were packed into a dense pellet by centrifugation, postfixed, then embedded in Epon. Mammalian eggs were not centrifuged; instead, they were embedded in agarose discs. Cells embedded in agarose were cooled below 30 degrees C to allow for gelling, then processed for electron microscopy. Because agarose has a low gelling temperature, some heat-labile substances were preserved, as demonstrated by retention of peroxidase activity using the DAB histochemical method. The agarose embedding procedure is both rapid and facile, and has proven to be of value in the handling of fragile single cells for electron microscopic studies.
Anat Rec 1981 Oct
PMID:An improved method for processing single cells for electron microscopy utilizing agarose. 703 63

Structural specializations in the plasma membrane of opossum spermatozoa obtained from different levels of the epididymis have been analyzed in thin sections and freeze-fracture replicas. The maturation process was accompanied by a redistribution of intramembranous particles in the flagellar midpiece region. Caput epididymal spermatozoa are immotile, and freeze-fracture replicas of the midpiece plasma membrane reveal a random arrangement of intramembranous particles. As spermatozoa transit the corpus epididymis, the intramembranous particles in the midpiece plasma membrane are redistributed from a random arrangement to an organized packing pattern. This redistribution apparently involves the formation of chains of intramembranous particles which gradually increase in length, orient parallel to the flagellar long axis, and ultimately form numerous parallel rows, each three to five particles wide. In cauda epididymal spermatozoa the intramembranous particles within the rows are packed in an organized manner, and few free intramembranous particles are noted between rows. Analysis of thin sections revealed that the reorganization of intramembranous particles is accompanied by the deposition of a mat of amorphous material at the cytoplasmic face of the membrane. No striking changes in intramembranous particle distribution during epididymal maturation were found in other flagellar segments or in the plasma membrane overlying the sperm head.
Anat Rec 1980 Aug
PMID:Changes in intramembranous particle distribution in the plasma membrane of Didelphis virginiana spermatozoa during maturation in the epididymis. 721 98

Membranes of boar spermatozoa from different regions of the epididymis and after ejaculation were studied by the freeze-fracture replica technique. The ordered pattern of the intramembranous particles of spermatozoan plasma membranes was different in the five arbitrary zones of the epididymis and in the semen. A distinctive ordered pattern was absent in zone 1, which is the proximal segment of the epididymis. In zone 2, paired parallel rows of the particles were present in the plasma membrane over the acrosomal region. This parallel arrangement was not present in zone 3 spermatozoa. Anterior to the posterior ring, cords formed by packed particles were apparent in zone 2 spermatozoa and reached their maximum prominence in zone 3, and persisted in zones 4 and 5 and in the semen. The plasma membrane over the marginal ridge of the acrosome had a hexagonal array of particles only in zones 4 and 5 spermatozoa. A similar pattern appeared on the post-acrosomal region of spermatozoa in zone 5 and in the semen. The plasma membrane of the middle piece had a rectilinear arrangement of the particles in zone 2 spermatozoa in which the migration of the cytoplasmic droplet was complete. Rudiments of the rectilinear arrangement persisted in spermatozoa in zones 4 and 5 and in the semen. These changes are discussed in relation to sperm maturation in the epididymis. The acrosomal membrane had a hexagonal arrangement of particles in the equatorial segment. The marginal ridge of the outer acrosomal membrane had parallel rows of intramembranous particles. The organization of the acrosomal membrane particles did not change during the epididymal passage of boar spermatozoa.
Anat Rec 1981 Mar
PMID:Changes in intramembranous particle distribution epididymal spermatozoa of the boar. 725 83

In the epididymis of the guinea pig, zone II exhibits striking histological features that distinguish it readily from the other six regions of the epididymis. At the light microscope level, the pseudostratified epithelium of zone II is characterized by tall principal cells that are densely packed with large, intensely staining granules or droplets ranging up to 8 mu in diameter. At the electron microscope level, the principal cells exhibit numerous large lipid droplets and abundant agranular endoplasmic reticulum, which is frequently arranged in concentric whorls around one or more of the droplets. Quantitative biochemical studies comparing zone II with zones I and III show that zone II contains 2.5 - 3-fold more cholesterol and a significantly greater amount of cholesterol ester than the other two zones. These data indicate that the epididymal duct of the guinea pig includes a clearly defined region of epithelial cells possessing ultrastructural and biochemical characteristics consistent with steroidogenic activity. The potential significance of these observations to the epididymal physiology of the guinea pig and epididymal physiology in general is discussed.
Anat Rec 1981 Dec
PMID:Studies on zonation in the epididymis of the guinea pig. I. Ultrastructural and biochemical analysis of the zone rich in large lipid droplets (zone II). 734 May 66

Our objective was to characterize epithelial cells, lamina propria, and sites of estrogen coupling in the caput, corpus, and cauda regions of the human epididymis using antibodies to cytokeratin types; epithelial membrane antigen; laminin; type IV collagen; vimentin; desmin-, and estradiol-receptor-related protein; and immuno-histochemical techniques. Principal cells immunostain by both AE1/AE3 antibodies (keratins 1-8, 10, 13-15, and 19) and anti-pan-keratin antibodies (keratin 5, 6, and 8). Immunoreactions to both anti-keratin antibodies increase from the caput to the cauda epididymis. The principal cells only immunostained by anti-keratin 19 antibodies in the cauda and showed no reaction to keratins 10 and 11. Basal cells and apical cells immunoreact to anti-AE1/AE3, antipankeratin, and antikeratin 19 antibodies, but not to antikeratin 10 and 11 antibodies, in all three epididymal regions. The principal cells immunoreact with epithelial membrane antigen antibodies in the stereocilia and subjacent cytoplasm. This immunostaining decreased from the caput to the cauda. Antivimentin antibodies stained the apical cytoplasm of principal cells and limited areas of both principal cells and basal cells. This immunoreaction decreased from the caput to cauda. Apical cells immunostained in the three regions. Immunoreaction to ER-D5 was moderate in the principal cells, basal cells, apical cells, and muscular coat cells in the cauda. The apical cells immunostained in the three regions. Antilaminin antibodies stained the epithelial basement membrane in the three regions. Type IV collagen was detected in the basement membrane as well as around the muscular coat cells in the three regions. Immunoreaction to desmin was intense in the muscular coat cells in the three regions.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1993 Apr
PMID:Immunohistochemistry of the human ductus epididymis. 768 39

To establish the mode of fertilization in a marsupial, a morphological investigation was made of the gametes of the South American grey short-tailed opossum. Monodelphis domestica, at the time of fertilization in vivo and in vitro. Oestrus was induced in females by the introduction of an unfamiliar male. To obtain oocytes recently fertilized in vivo, females were killed 18-24 hours after the first mating and the region of the oviduct containing eggs excised and fixed. Unfertilized mature oocytes were recovered from ovarian follicles 15-18 hours after first mating and fertilized in vitro with cauda epididymal spermatozoa in a modified MEM medium supplemented with bovine serum albumin at 37 degrees C in 5% CO2 in air. Following sperm-egg binding and fertilization, oocytes were fixed and prepared for light and electron microscopy. Spermatozoa unpaired prior to fertilization in vivo and in vitro and single spermatozoa bound to the zona surface by their plasmalemma overlying the acrosome on the dorsal face of the sperm head. The acrosome reaction was only observed at the zona surface (suggesting that it may be induced by zona components) and involved a vesiculation of sperm plasma and acrosomal membranes over the main body of the acrosome but not over the narrow, marginal region which persisted after the acrosome reaction was complete. Sperm penetration of the zona pellucida caused a large breach in the zona and the dispersal of perivitelline material. The fusion of the spermatozoon with the oolemma occurred first over the marginal acrosomal region and was accompanied by a fertilization cone which protruded through the zona penetration hole. Activation of the egg was characterized by the release of material from vesicles in the peripheral cytoplasm and extrusion of the second polar body. The mode of fertilization in Monodelphis was compared with what is known in other marsupials (New World and Australian) and eutherian (placental) mammals. It was concluded that the general features of the acrosome reaction and sperm-egg fusion may be essentially similar in both groups and that an evolutionary schism did not occur following the development of the eutherian mode of fertilization.
Anat Rec 1993 Sep
PMID:Ultrastructural characteristics of in vivo and in vitro fertilization in the grey short-tailed opossum, Monodelphis domestica. 821 40


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