Rat Testicular Dolichols: Distribution and Accumulation with Age1

Nyquist, S. E.; Holt, Sidonie

doi:10.1095/biolreprod34.1.89

Cited by 19 publications

(8 citation statements)

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“…This agrees with earlier work from this laboratory (Nyquist and Holt 1986), in which it was reported that spermatogenic cells contain very low dolichol concentrations. Additionally, the biosynthetic data suggest that Sertoli cells synthesize dolichols, under the conditions used in these experiments, more effectively than do the spermatocytes.…”

Section: Discussionsupporting

confidence: 93%

“…This interpretation was further supported by studies of Potter et al (1980), in which mouse pachytene spermatocyte fractions, following [14c]acetate incorporation, were isolated by unit gravity sedimentation and observed to have the highest [14c]acetate incorporation into dolichol. Analysis of the dolichol content of pachytene spermatocyte fractions (Nyquist and Holt 1986), however, showed a relatively low concentration, with most of the dolichol being recovered in fractions containing the Sertoli cells.…”

Section: Introductionmentioning

confidence: 92%

“…Pachytene spermatocytes were isolated as described previously by Nyquist and Holt (1986) using a Beckman JE-1OX elutriator and a Beckman 52-21 centrifuge. Aseptic technique was employed during all stages of the cell separations.…”

Section: Centrifugal Elutriation Of Pachytene Spermatocytesmentioning

confidence: 99%

“…Although N-linked glycoprotein biosynthesis is a very active process in the seminiferous tubules and dolichols are present in abundance (Nyquist and Holt 1986;Keller and Nellis 1986), very little is known about the distribution of these compounds within the testis, their biosynthetic pathway, or their role in the regulation of glycoprotein synthesis during spermatogenesis. The process of spermatogenesis is glycoprotein dependent; the formation of the acrosome involves extensive synthesis of glycoproteins and stage-specific spermatogenic cell surface glycoproteins have been demonstrated (Millette and Scott 1984;Gerton and Millette 1986).…”

Section: Introductionmentioning

confidence: 99%

“…Of the nonphosphorylated dolichols, Tollbom et al (1988) report that 38.5% exists as dolichyl esters. Testicular Do1 concentrations increase substantially with age (Nyquist and Holt 1986); Dol-P increases with age, but to a much lesser degree, (i.e., two-to three-fold) (Keller and Nellis 1986;Chen et al 1988).…”

Section: Introductionmentioning

confidence: 99%

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Dolichol and N-linked oligosaccharide synthesis in the rat testis: interaction between Sertoli and spermatogenic cells, evidence for paracrine effects

Page

Mason²,

Lindstrom³

et al. 1992

Biochem. Cell Biol.

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The relative contribution of the Sertoli cell and the pachytene spermatocyte to dolichol and N-linked oligosaccharide biosynthesis within the seminiferous tubule was investigated. Evidence is presented to show that the interaction between these two cell types affects dolichol and N-linked oligosaccharide biosynthesis. Analysis of the dolichol content of Sertoli cultures confirms earlier data suggesting that the Sertoli cell constitutes the major pool of dolichols within the seminiferous tubule. [14C]Acetate incorporation studies suggest that the Sertoli cell in culture synthesizes dolichol much more rapidly than does the isolated pachytene spermatocyte. This information, in addition to previous data in the literature, infers an interactive effect whereby the presence of the spermatogenic cell in the tubule stimulates dolichol synthesis in the Sertoli cell. The absence of normal Sertoli-spermatocyte interactions in in vitro incubations may also limit dolichol synthesis in the pachytene spermatocyte. The distribution of dolichol kinase between the Sertoli and the pachytene spermatocyte was also examined. The concentration of this enzyme in the Sertoli cell suggests the presence of an active salvage pathway within that cell. The correlation between the appearance of the pachytene spermatocyte and the previously described peak of dolichol kinase activity in the seminiferous tubules of the prepubertal animal implies cell-cell interactions. Radiolabelling studies of N-linked oligosaccharides were conducted using [3H]mannose and concanavalin A affinity chromatography to identify multiantennary, biantennary, and high-mannose oligosaccharide pools. An in vitro bicameral coculture system was used to demonstrate that pachytene spermatocytes stimulate incorporation of [3H]mannose into Sertoli cell oligosaccharides.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 92%

Section: Centrifugal Elutriation Of Pachytene Spermatocytesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Dolichol and N-linked oligosaccharide synthesis in the rat testis: interaction between Sertoli and spermatogenic cells, evidence for paracrine effects

Page

Mason²,

Lindstrom³

et al. 1992

Biochem. Cell Biol.

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Characterization of Membrane Rafts Isolated From Rat Sertoli Cell Cultures: Caveolin and Flotillin‐1 Content

Evans

Coyer

Sandusky

et al. 2003

Journal of Andrology

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Membrane rafts from Sertoli cell cultures were isolated as detergent-insoluble glycosphingolipid-enriched (DIG) fractions on the basis of their enriched content of glycosphingolipids and cholesterol and the resulting insolubility in 1% Triton X-100 and their low buoyant density. Because lipid rafts have been implicated in numerous cell functions, including cell signaling and sites for actin/membrane attachment, studies were initiated to characterize Sertoli cell rafts. This study reports the distribution of the raft structural proteins, caveolin and flotillin-1, implicated in raft microdomain organization. Methods employed included the immunoblotting of cell lysates and detergent-insoluble glycosphingolipid-enriched (DIG) fractions, the immunofluorescent microscopy of peritubular myoid cell (PMC) cultures and cryostat-sectioned testis, and the immunohistochemical staining of paraffin-embedded sections following microwave antigen retrieval techniques. Sertoli cells and Sertoli DIG fractions were found to lack the common raft-associated protein, caveolin, a marker protein for caveolae, but they are enriched in the 48-kd protein, flotillin-1, a protein also implicated in raft formation, cell signaling, and cell motility. Since the primary cell contaminant of Sertoli cell cultures is the PMC, these cells, along with spermatogenic cell fraction (SPGC), were also examined for caveolin and flotillin-1 content. The PMCs contained significant concentrations of both caveolin and flotillin-1. PMCs in culture exhibited a punctate caveolin staining pattern at the cell surface characteristic of a caveolar location. These data support the idea that the pinocytotic vesicles observed in PMCs are caveolae. PMCs also show a perinuclear location for caveolin characteristic of a Golgi location. Cryostat sections of rat testis showed a marked concentration of caveolin in the PMCs. The PMC location of caveolin was also confirmed by the immunohistochemical staining of sections from paraffin-embedded rat testis following microwave antigen retrieval techniques. Similar experiments showed a more ubiquitous, stage-specific distribution of flotillin-1 among testicular cell types.

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