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Biomedical subjects

L D Russell

Publications and source records attributed to L D Russell.

At least 127 records · Page 7Linked to original sources
PubMed1983

Penetration of zona-free hamster eggs by boar sperm treated with the ionophore A23187 and inhibition of penetration by antiplasma membrane antibodies.

Uncapacitated boar sperm treated with the divalent cation ionophore A23187 in the presence of calcium bind to and penetrate zone-free hamster eggs. Penetration by ionophore-treated sperm is, however, blocked when sperm are pretreated with antisperm plasma membrane Fab. The antibody apparently interferes with the interaction of sperm surface antigens with the egg plasma membrane since the antibodies have no significant effect on the motility of sperm nor on the induction of the acrosome reaction by the ionophore. The assay described herein is useful for studying the role of sperm surface antigens in the penetration and fertilization of eggs.

Animals↗
PubMed1983

Development of the acrosome and alignment, elongation and entrenchment of spermatids in procarbazine-treated rats.

Intraperitoneally administered procarbazine caused, among other features previously reported (Russell et al., 1983), specific defects in the acrosome of cap phase spermatids of the rat seminiferous epithelium. The effect of procarbazine was to fragment and eventually cause resorption of the acrosomes of a small number of steps 5--9 spermatids. Although the acrosome was lost, close union of the leaflets of the nuclear envelope underlying the acrosomal sac was maintained as was the marginal fossa and acrosomal zonule. Spermatids at steps 8 and 9 of development, which had lost their acrosomes, showed nuclei which were eccentric within the cell--a feature which normally occurs at these steps of spermiogenesis in acrosome intact cells. Even without an acrosomal sac, the plasma membrane of these cells (in stage VIII) became orientated to the region of the nuclear membrane which would have underlaid the acrosome. Although abundant, Sertoli ectoplasmic specialization did not become aligned with the spermatid head. The spermatid failed to become orientated within the seminiferous epithelium and failed to enter the crypts within the Sertoli cell as usually occurs during the elongation process. Thus, the presence of an acrosome is not likely related to the formation of an eccentric nucleus or the alignment of the surface of the nucleus which would normally underlay the acrosome with the cell's plasma membrane (internal alignment). The presence of an acrosome may be related to the alignment of the spermatid head with the ectoplasmic specialization, which in turn may influence the orientation and positioning of the late spermatids within the seminiferous epithelium (external alignment) and their position within recesses of the Sertoli cell. This study also suggests a role for the manchette in the process of elongation of the spermatid.

Acrosome↗
PubMed1983

Morphological pattern of response after administration of procarbazine: alteration of specific cell associations during the cycle of the seminiferous epithelium of the rat.

Procarbazine, an anti-cancer agent, administered intraperitoneally to adult, male rats induced a characteristic morphological pattern of response in the seminiferous epithelium. Seminiferous tubules of rats receiving 100 mg/kg procarbazine and higher dosages displayed spermatids which were less mature than those normally found within seminiferous tubules which show a particular cell association. Early spermatids in steps 1-7 of spermiogenesis appeared arrested in their development and were present in cell associations which had advanced normally. The most probable cause of this apparent germ cell arrest was a retardation of acrosomal development since procarbazine is known to affect RNA and consequently protein synthesis. Other features which indicated defective RNA synthesis were the presence of abnormal spermatid nucleoli and abnormally configured chromatoid bodies. This study demonstrates, in contrast to what is indicated by present dogma, that apparent and temporary germ cell arrest may occur under certain deleterious conditions. It also illustrates that particular cell types within a cell association may be out of synchrony with the remainder of the cells in a cell association.

Animals↗
PubMed1983

Electrophoretic map of boar sperm plasma membrane polypeptides and localization and fractionation of specific polypeptide subclasses.

A high resolution, two-dimensional (2-D) electrophoretic map of the plasma membrane (PM) polypeptides from the ejaculated boar spermatozoon is described. 2-D silver-stained polyacrylamide gel electrophoresis (PAGE) gels revealed over 250 polypeptides; Coomassie blue staining revealed more than 100. Fifty Coomassie-staining polypeptides were catalogued and biochemically characterized, with twenty of these designated major sperm PM polypeptides. Cavitation pressures ranging from 50 PSI to 1000 PSI were used to enrich 2-D maps either in head PM (50 PSI) or in flagellar PM (1000 PSI) and provided tentative localization of certain PM polypeptides. Immunoabsorption chromatography showed that most major polypeptides seen in the 2-D map were antigenic. Major polypeptide bands from single dimensional (1-D) gels were excised, antibodies against them were produced in rabbits, and the polypeptides were localized via indirect fluorescein isothiocyanate (FITC) technique. Cross-reacting antigenic determinants in the PAGE PM polypeptides were determined by transblotting and staining the transblots by an indirect peroxidase technique. Cross-reactivity was extensive with several polypeptide groups, but specific enough with others to allow tentative localization. Lectin affinity chromatography using Con A, WGA, RCA-1, PNA, and DBA indicated the lectin specificity of PM polypeptides. These data together with periodic acid-Schiff (PAS) and carbohydrate-specific silver staining permitted identification of glycoproteins in the 2-D maps. FITC coupled to specific lectins showed the regional distribution of these lectins on the sperm surface. The 2-D polypeptide map and the catalogue of properties of major Coomassie-stained PM polypeptides provides a reference for future studies in the boar and other species.

Animals↗
PubMed1982

The effects of antisperm plasma membrane antibodies on sperm-egg binding, penetration, and fertilization in the pig.

A procedure using published surgical techniques is described for determining the effects of antiplasma membrane antibodies on sperm-egg binding, penetration, and fertilization in vivo in the domestic pig. Time of ovulation was controlled and sperm inseminated at precise times relative to ovulation. Sperm were exposed to antibodies (Fab) and then washed free of excess antibody and placed in one uterine horn near the uterotubal junction. Sperm exposed to control material were placed in the opposite horn as a control. One horn was tied with two ligatures near the body of the uterus. Eggs were collected by flushing the oviducts at prescribed times after insemination. in nine animals used for this study, antibodies to boar sperm plasma membranes completely blocked sperm-zona binding, penetration of the zona, and fertilization. In contrast, sperm were bound to the zona pellucida and penetrated eggs in every case on the control side. These results indicate that insemination in vivo is useful in screening antibodies produced against specific plasma membrane antigens to determine their role in fertilization.

Animals↗
PubMed1982

Visualization of anti-sperm plasma membrane IgG and Fab as a method for localization of boar sperm membrane antigens.

A simple method for ultrastructural localization of sperm surface antigens by direct visualization of bound antibodies is presented. Anti-sperm plasma membrane (ASPM) immunoglobulin (Ig) G, visualized in tissues treated with an osmium:ferrocyanide mixture, projected 11-13 nm from the surface and ASPM Fab fragments projected 8-10 nm from the surface. The density of IgG labeling, as subjectively estimated, corresponded to indirect immune fluorescein isothiocyanate, indirect immunoferritin, and sperm-vesicle labeling patterns. Agglutination of sperm vesicles and sperm were demonstrated and the linking antibody visualized. A second antibody on protein A directed against ASPM IgG made the immunologic tag more apparent and indicated, in disrupted sperm preparations, labeling of both sides of the plasma membrane. The method provides for easy and sensitive localization of sperm surface antigens at the ultrastructural level and is presently being used to localize specific sperm antigens.

Animals↗
PubMed1981

Effect of the microtubule disrupting agents, colchicine and vinblastine, on seminiferous tubule structure in the rat.

Injections of colchicine or vinblastine were given intratesticularly and rats sacrificed 6 and 12 hr later. Colchicine and vinblastine produced identical morphological patterns of response in the seminiferous tubules resulting in arrest of germcell mitoses and meioses and a rapid depletion of the microtubules normally found within the Sertoli cell. Sloughing of cells into the lumen of seminiferous tubules was the most prominent feature noted. Germ cells and portions of the apical Sertoli cells were frequently sloughed together where they remained in close association. Usually germ cells and associated Sertoli cell fragments were cleaved from the wall of the seminiferous tubule at a level between dissimilar generations of germ cells, e.g. between spermatocytes and spermatids. This selective sloughing probably occurred as the result of the support normally provided by intercellular bridges which link clones of like germ cell types. Sequential steps in the process leading to sloughing of Sertoli-germ cell associations could be inferred from observations made in plastic 1 micrometer sections. Cell sloughing at 12 hr post-injection was generally more extensive. It was frequently noted that germ cells and the apical portions of Sertoli cells had been extruded to the level of the most adluminal tight junctions forming the blood-testis barrier. It was concluded that disruption of Sertoli microtubules was responsible for sloughing of Sertoli fragments and associated germ cells, and that the cytoskeletal support, of the Sertoli cell was, at least in part, dependent upon the integrity of Sertoli microtubules. The Sertoli cell could not round-up after loss of its cytoskeletal support, due to the numerous attachment devices known to link it with various apically positioned germ cells. Thus, the cell was severed at some point along its delicate apical processes, as the consequence of forces produced by the 'rounding-up' process. Long-term sacrifice after vinblastine or colchicine treatment allowed the Sertoli cells to regain microtubules and long processes but not their typical configuration. Spermatogenesis remained severely impaired.

Animals↗
PubMed1981

Morphological pattern elicited by agents affecting spermatogenesis by stimulation.

A variety of agents (clomiphene citrate, cyproterone acetate, estradiol 17-beta, medrogestone, medroxyprogesterone, and methoxychlor) thought to disrupt the normal pathway by which hormones stimulate the testis were tested in rats to determine the early morphologic changes in the testis. After initial trial experiments at which dose and sacrifice intervals were determined, the earliest morphological abnormalities were assessed quantitatively and related to the particular spermatogenic stage in which these defects were initiated. All agents tested produced an identical morphological pattern of response displaying a significant increase in Stage VII degenerating cells (pachytene spermatocytes, step 7 and step 19 spermatids) as compared with vehicle-administered rats. The number of degenerating cells in other stages did not significantly change. The rise in degenerating cells in Stage VII, as seen in this study, was similar to that reported in Russell and Clermont (1972) utilizing hypophysectomized rats, and that reported by other investigators who have used a variety of agents to disrupt the hormonal stimulation of the seminiferous tubules. The data indicate that no matter how, or at what level the hormonal stimulation of the testis is interrupted, the morphological pattern of response is the same, and that such a pattern is useful in predicting the mechanism of action of agents suspected of interfering with hormone action.

Animals↗
PubMed1981

Spermatophores of the salamander Ambystoma texanum.

Living spermatozoa were observed in freshly deposited spermatophores and in fluid from vasa deferentia. In the distal, but not proximal, vas deferens spermatozoa moved together in whorls with heads and tails in alignment. Around the entire periphery of the spermatophore cap, similar slowly undulating groups of spermatozoa had their heads aligned and directed outward. Over time, some individual spermatozoa left the cap of the spermatophore and moved into the surrounding water (cap deterioration). Microscopical observations were made on spermatophore squashes and paraffin sections of spermatophores and vasa deferentia. Spermatozoa around the periphery of the cap were underlain by a PAS-positive membrane-like material. Cytoplasmic droplets, which were attached to spermatozoan necks in the vas deferens, were accumulated in the center of the spermatophore cap deep to the PAS-positive membrane. Spermatophore stalks were strongly PAS and Alcian blue positive and showed positive reaction for RNA. Tests for lipids and proteins were negative in the whole spermatophore. Electron microscopic observations showed the stalk of the spermatophore to be composed of rounded 'balls' of fibrous material. At the juncture of the stalk and cap a less dense fibrous material impacted the stalk enclosing many sperm tails and some heads and, although no attachment devices were visualized, the sperm were closely apposed to this material as was the spermatophore stalk. This finely filamentous material encircled the cap and was more prominent in some regions than others. The PAS-positive material detected with the light microscope was also observed with the electron microscope. It was circumferentially oriented and was composed of 200 A packed filamentous densities. Sperm heads and tails were found lying external to the membrane, whereas only tails and cytoplasmic droplets occupied the core of the spermatophore. Cytoplasmic droplets were usually free of the sperm tail and contained membranous sacs and two types of nuage material.

Ambystoma↗
PubMed1980

Deformities in the head region of late spermatids of hypophysectomized-hormone-treated rats.

Testosterone capsules implanted in hypophysectomized rats prevented tubulobulbar complexes from forming in many late spermatids. These spermatids also displayed an accumulation of cytoplasm (swelling) in the perinuclear region of the head. Other spermatids with normal or near-normal numbers of tubulobulbar complexes showed a typical perinuclear space. The results indicate, but do not prove, that development of tubulobulbar complexes (and their subsequent phagocytosis) is essential for the normal elimination of cytoplasm that takes place prior to sperm release.

Animals↗
PubMed1980

A study of Sertoli-spermatid tubulobulbar complexes in selected mammals.

Tubulobulbar complexes (TBCs) were found in nine mammalian species (opossum, vole, guinea-pig, mouse, hamster, rabbit, dog, monkey and human) primarily originating from the plasma membrane overlying the acrosome of late spermatids. Fewer complexes (4--10) were noted in these species than has been previously reported for the rat (up to 24). TBCs were not seen emanating from round spermatids or those elongated spermatids located within the deep recesses of the Sertoli cell, but they appeared as the spermatids came to reside much closer to the tubular lumen in preparation for release. TBCs developed in areas deficient or lacking in Sertoli filaments and endoplasmic reticulum (ectoplasmic specialization). In general their structural configuration was similar to that shown in the rat, although minor differences were noted. Fine fibrils were observed connecting the distal portion of the spermatid tube with the Sertoli plasma membrane forming a bristle-coated pit. The length of TBCs from most species studied was 1--2 micrometers, whereas those of the opossum extended 6--8 micrometers into an apical Sertoli process. TBCs were degraded within the Sertoli cell by its lysosomes prior to sperm release, and for most species there was evidence indicating that formation of more than one generation of TBCs occurred. As sperm release approached, TBCs formed preferentially from the leading edge of spermatids with spatulate heads. The Sertoli cell gradually withdrew from around the spermatid head until only the tip of the head was embedded within the Sertoli cell. This region of contact frequently demonstrated TBCs. The proposed functions of TBCs are reviewed and discussed in light of these findings from other species.

Animals↗
PubMed1980

Morphological observations on the binding of boar sperm to porcine zona pellucida.

Scanning electron microscopy demonstrated that freshly ejaculated boar sperm bind in large numbers to the cumulus-free zona pellucida of porcine oocytes. Sperm were bound head-first although some portions of the flagellum occasionally made contact with the zona. Scanning microscopy also showed that sperm did not undergo an acrosome reaction or penetrate the zona after binding. Binding of boar sperm was visibly inhibited by pretreatment of the sperm by Fab antibodies to boar sperm plasma membranes. The forgoing indicate that specific receptors for the zona pellucida are present and exposed on the surface of non-capacitated boar sperm. In addition, when soluble components which were extracted from sperm by sedimentation at high speed, were preincubated with oocytes, sperm did not bind. Minor surface changes were noted after these treatments, but the zona remained intact and was not penetrated by sperm. Either this treatment inactivated sperm receptors or the outer layer of the zona, which may be the only portion of the zona containing receptors, was removed. Since binding of the sperm to the zona pellucida involved the sperm plasma membrane, we suggest that this membrane is intact when the sperm reaches the zona. Surface changes accompanying capacitation are apparently not necessary for species-specific sperm-egg binding in vitro in the pig.

Animals↗
PubMed1979

Observations on the inter-relationships of Sertoli cells at the level of the blood- testis barrier: evidence for formation and resorption of Sertoli-Sertoli tubulobulbar complexes during the spermatogenic cycle of the rat.

Structures termed tubulobulbar complexes are known to be formed by adjoining Sertoli cells at the level of the blood-testis barrier (Russell and Clermont, '76). Here, long (2-4 micrometer) tubular evaginations of one Sertoli cell, which end in bulbous dilations, are seen in corresponding invaginations of a neighboring Sertoli cell. In most regions of the tubular and bulbous portions of the complex, the Sertoli plasma membranes were found to be separated by a 4-5-nm intercellular space, but in some areas the membranes converged to form tight and gap junctions. The numbers, distribution and properties of tubulobulbar complexes were studied in relation to the cycle of the seminiferous epithelium. From the data obtained it was concluded that tubulobulbar complexes develop and undergo regressive changes during the spermatogenic cycle. Most complexes arise during the early stages of the cycle (Stages II-V) and develop large bulbous endings. Developing tubulobulbar complexes consist of short evaginations of one Sertoli cell which face a bristle-coated pit of the opposing Sertoli cell. At midcycle (Stages VI-VII) most show regressive changes and are eventually resorbed as a consequence of the action of nearby Sertoli lysosomes. Once resorbed, the probability of seeing a tubulobulbar complex in thin sections decreases from 4- to 8-fold. The few tubulobulbar complexes which remain past this period (Stages VII-XIV-I) usually lack bulbous endings and are fequently seen above type A spermatogonia. The data suggest that small fragments of cytoplasm and plasma membrane (including junctional surfaces) are lost from one Sertoli cell as a result of the degradative processes occurring in a neighboring Sertoli cell. Tubulobulbar resorption is discussed in relation to the impending breakdown of the blood-testis barrier above spermatocytes as these cells move upward. The possible significance of the cyclic resorption of tight and gap junctional sites between Sertoli cells is also discussed.

Acid Phosphatase↗
PubMed1978

The blood-testis barrier and its formation relative to spermatocyte maturation in the adult rat: a lanthanum tracer study.

An electron-opaque substance, lanthanum, was utilized to determine when germ cells of the rat first cross the blood-testis barrier in adult spermatogenesis. Intravascularly perfused lanthanum was shown to surround all spermatogonia. Intravascularly perfused lanthanum was shown to surround all spermatogonia, preleptotene spermatocytes and early leptotene spermatocytes of Stage IX in the adult rat testis. Lanthanum was excluded from the spaces around more mature cells by newly-formed tight junctions between adjoining Sertoli processes. These processes had previously intervened between the leptotene cells and the basal lamina. The results are in close agreement with those of a previous study (Russell, '77a) which indicated that leptotene cells are the first cells of adult spermatogenesis to enter the intermediate compartment and to reside beyond a permeability barrier.

Animals↗