Spatially dynamic intercellular adhesion junction is coupled to a microtubule-based motility system: Evidence from an in vitro binding assay

Vogl, A. Wayne

doi:10.1002/(sici)1097-0169(1996)34:1<1::aid-cm1>3.0.co;2-g

Cited by 23 publications

(19 citation statements)

References 43 publications

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“…The actin filaments are hexagonally packed [3] and uniformly polar [4], and are not associated with myosin II [5]. Intact ectoplasmic specializations remain attached to spermatids that have been mechanically dissociated from the seminiferous epithelium [6]. This result not only indicates a strong attachment between Sertoli cells and spermatids in regions of the junction plaque, but it is also consistent with the conclusion that the various components of the plaque function as a structural unit.…”

Section: Introductionsupporting

confidence: 58%

“…Also, the microtubules are closely associated with apical crypts containing attached spermatids and are specifically related to the specialized adhesion junction plaques surrounding the crypts [1]. Moreover, spermatid/junction complexes similar to those used in the motility assay bind microtubules in a nucleotide-dependent fashion [6,13], an observation that is consistent with the presence of motor proteins on the plaques.…”

Section: Discussionmentioning

confidence: 85%

“…A fraction enriched in spermatids was obtained by centrifuging the epithelial material through a step sucrose gradient. Previous studies have demonstrated that intact ectoplasmic specializations remain attached to spermatids obtained in this fashion [6]. The spermatids with associated junction plaques were then mounted in simple flow chambers and assayed for their ability to transport fluorescently labeled microtubules.…”

Section: Methodsmentioning

confidence: 99%

“…A testicular fraction enriched for elongate spermatids with attached junction plaques was prepared using a protocol similar to one used previously to obtain material for a microtubule binding assay [6]. Animals were anesthetized with halothane, and their testes were removed and decapsulated.…”

Section: Preparation Of Elongate Spermatids With Attached Junction Plmentioning

confidence: 99%

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Spermatid Translocation in the Rat Seminiferous Epithelium: Coupling Membrane Trafficking Machinery to a Junction Plaque1

1999

Biology of Reproduction

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In this study, we demonstrate that specialized junction plaques that occur between Sertoli cells and spermatids in the rat testis support microtubule translocation in vitro. During spermatogenesis, Sertoli cells are attached to spermatids by specialized adhesion junctions termed ectoplasmic specializations (ESs). These structures consist of regions of the plasma membrane adherent to the spermatid head, a submembrane layer of tightly packed actin filaments, and an attached cistern of endoplasmic reticulum. It has been proposed that motor proteins on the endoplasmic reticulum interact with adjacent microtubules to translocate the junction plaques, and hence the attached spermatids, within the epithelium. If this hypothesis is true, then isolated junctions should support microtubule transport. To verify this prediction, we have mechanically isolated rat spermatids, together with their attached ESs, and tested them for their ability to transport microtubules in vitro. Most assays were done in the presence of 2 mg/ml testicular cytosol and at room temperature. ESs attached to spermatids supported microtubule translocation. In some cases in which motility events were detected, microtubules moved smoothly over the junction site. In others, the movement was slow but progressive, saltatory and "inch-worm-like." No motility was detected in the absence of exogenous ATP or in the presence of apyrase (an enzyme that catalyses the breakdown of ATP). Our results are consistent with the microtubule-based motility hypothesis of spermatid translocation.

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Section: Introductionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 85%

Section: Methodsmentioning

confidence: 99%

Section: Preparation Of Elongate Spermatids With Attached Junction Plmentioning

confidence: 99%

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Spermatid Translocation in the Rat Seminiferous Epithelium: Coupling Membrane Trafficking Machinery to a Junction Plaque1

1999

Biology of Reproduction

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“…Sloughing of germ cells was explained by others on the basis of organization of germ cells that are held in place by a close association between their plasmalemmas and specialized junctions of cell membranes of Sertoli cells. Early signs of cellular degeneration of germ cells might lead into disturbance of structure of their plasmalemmas causing their shedding into the lumina of seminiferous tubules [38,42,43].…”

Section: Discussionmentioning

confidence: 99%

Light and electron microscopic study on the effect of antischizophrenic drugs on the structure of seminiferous tubules of adult male albino rats

Soliman

Wagih

Attia

et al. 2015

Folia Histochem Cytobiol.

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Introduction. Sexual dysfunction and infertility are symptoms which have been rarely studied in patients treated with antischizophrenic drugs, aripiprazole and olanzapine, for long period. This work aimed to investigate the effects of aripiprazole and olanzapine on the structure of seminiferous tubules of rats at both light microscopic and ultrastructural levels. Material and methods. Sixty adult male rats were divided into 3 groups (n = 20): control group (Group I) and two experimental ones (II and III). Rats in Group II received 2 mg/kg/day aripiprazole while rats in Group III received 0.5 mg/kg/day olanzapine for 14 weeks. Thereafter, testis were removed and processed for both light and electron microscopic study. Qualitative morphological analyses and histomorphometric measurements of seminiferous tubules were performed. Results. Rats in Group II showed reduction of testicular weight, seminiferous tubules' diameter, epithelial height, spermatogenic count, spermatogenic index and spermatogenic score whereas Sertoli cells count was increased. Olanzapine-treated rats also showed epithelial desquamation, separation and apoptotic changes of germ cells. Sertoli cells showed vacuolization, dilatation of smooth endoplasmic reticulum and accumulation of lipid droplets. Abnormality in the shape and structure of late spermatids and presence of giant cells were also demonstrated. Aripiprazole induced less adverse histological changes in rat testis than olanzapine. Conclusions. Olanzapine followed by aripiprazole had adverse histological effects on the structure of the seminiferous tubules, which may affect spermatogenesis.

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A re‐evaluation of gelsolin at ectoplasmic specializations in sertoli cells: The influence of serum in blocking buffers on staining patterns

Guttman

Vaid

Vogl

2007

The Anatomical Record

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In this study, we test the hypothesis that gelsolin immunolocalized in actin filament-rich ectoplasmic specializations may be exogenous gelsolin present in normal serum used in blocking buffers, and that binds to the intercellular adhesion plaques during tissue processing. Fixed frozen sections of rat and rabbit testis were pre-treated with standard blocking buffers containing 5% normal goat serum (NGS) and then incubated with anti-gelsolin antibodies in the presence of 1% NGS. Other sections were treated in a similar fashion, but in buffers not containing NGS. Sections were then labeled with secondary antibody conjugated to a fluorochrome. Localized staining at ectoplasmic specializations occurred only in sections treated with NGS. The only positive staining in sections not treated with NGS was associated with seminiferous tubule walls and blood vessels in rabbit tissue. The antibodies reacted with a single band at the appropriate molecular weight for gelsolin on immunoblots of NGS, but did not react on immunoblots of testis or seminiferous epithelium. We conclude that gelsolin localized at ectoplasmic specializations using current commercially available antibodies is a result of non-specific binding to the fixed tissues of gelsolin present in blocking buffers. Anat Rec, 290:324-329, 2007. 2007 Wiley-Liss, Inc.Key words: gelsolin; ectoplasmic specializations; serum; testis; Sertoli Sertoli cells are attached to neighboring cells by elaborate intercellular adhesion complexes termed ''ectoplasmic specializations.'' These structures are characterized by the Sertoli cell plasma membrane, a layer of actin filaments and an attached cistern of endoplasmic reticulum. In apical regions of the epithelium, ectoplasmic specializations function to attach spermatids to the seminiferous epithelium and their disassembly is part of the mechanism of sperm release. In basal regions of the epithelium, ectoplasmic specializations associate with other junction types (tight and gap junctions, desmosomes) to form a large junction complex. Tight junctions in this complex form the blood-testis or Sertoli cell barrier that divides the epithelium into a small basal compartment below the junctions and a large adluminal compartment above. At basal sites, ectoplasmic specializations and other components of the junction complex disassemble above spermatocytes and reassemble below as these cells move from basal to adluminal compartments of the epithelium. There currently is interest in the assembly, disassembly, and regulation of ectoplasmic

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Spatially dynamic intercellular adhesion junction is coupled to a microtubule-based motility system: Evidence from an in vitro binding assay

Cited by 23 publications

References 43 publications

Spermatid Translocation in the Rat Seminiferous Epithelium: Coupling Membrane Trafficking Machinery to a Junction Plaque1

Spermatid Translocation in the Rat Seminiferous Epithelium: Coupling Membrane Trafficking Machinery to a Junction Plaque1

Light and electron microscopic study on the effect of antischizophrenic drugs on the structure of seminiferous tubules of adult male albino rats

A re‐evaluation of gelsolin at ectoplasmic specializations in sertoli cells: The influence of serum in blocking buffers on staining patterns

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