Late spermatocytes from immature rat testis isolation, electron microscopy, lectin agglutinability and capacity for glycoprotein and sulfogalactoglycerolipid biosynthesis

Letts, Pamela J.; Hunt, Richard C.; Shirley, Margaret A.; Pinteric, L.; Schachter, Harry

doi:10.1016/0304-4165(78)90267-2

Cited by 22 publications

(9 citation statements)

References 27 publications

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“…Previous studies have shown that the synthesis of SGG during the first wave of spermatogenesis occurs, in the rat, between 15 and 20 d after birth, corresponding to the appearance of early primary spermatocytes (12). However, purified preparations of pachytene spermatocytes do not show an enhanced rate of SGG synthesis when compared with the total cell complement (14), which implies that SGG is synthesized before the pachytene spermatocyte stage. The present experiments have shown by both immunofluorescence (Fig .…”

Section: Discussionmentioning

confidence: 90%

“…Pachytene spermatocytes were prepared from the testes of 20 23-d-old rats and round (early) spermatids were prepared from two adult rat testes . Testes were stripped of their tunica and subjected to mechanical dissociation using a razor blade as previously described (14) . The mince was suspended in 20 ml of enriched Krebs-Ringer's bicarbonate buffer (EKRB) as described by Bellvt; et al .…”

Section: Cell Purificationmentioning

confidence: 99%

“…Fractions were centrifuged for 10 min at 800 g to sediment cells. A portion of these cells was suspended in buffer and examined with a phase-contrast microscope; cells were also examined by electron microscopy as previously described (14) . The pachytene spermatocyte preparation was 82% enriched and the spermatid preparation was 90% enriched according to electron microscopy criteria.…”

Section: Cell Purificationmentioning

confidence: 99%

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Localization of sulfatoxygalactosylacylalkylglycerol at the surface of rat testicular germinal cells by immunocytochemical techniques: pH dependence of a nonimmunological reaction between immunoglobulin and germinal cells.

Lingwood¹,

Schachter²

1981

The Journal of Cell Biology

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The synthesis of sulfatoxygalactosylacylalkylglycerol (SGG) is a marker of germinal cell differentiation during spermatogenesis . Antibodies raised against this lipid have been used to visualize SGG on the surfaces of rat spermatocytes and spermatids . An ionic interaction between SGG and immunoglobulin was shown to occur at physiological pH, resulting in high fluorescence backgrounds for control cells treated with nonimmune sera . Immunofluorescence was therefore performed at alkaline pH such that this interaction was much reduced or eliminated . A method was also developed to detect surface-bound complement fixed in the presence of anti-SGG . SGG was found to be mobile within the plane of the membrane, undergoing ligand-induced "patching" and occasional "capping ." However, this phenomenon was independent of temperature.Male germ cell differentiation offers a unique opportunity to study the biochemistry of differentiation . The strict synchrony of events, dramatic structural reorganization (28) and implicit role of intercellular communication make spermatogenesis an ideal system for the study of cell membrane differentiation . 1-0-hexadecanyl-2-0-hexadecanoyl-3-0-,8-[galactopyranosyl(3-0-sulfate)]-glycerol (sulfatoxygalactosylacylalkylglycerol, SGG)' is the major glycolipid of mature testes from all mammalian species studied so far (7,13,16,21). It has been shown in SGG both from the rat (12) and from the pig (7) that >90% of the constituent fatty acids were palmitic and >80% of the glyceryl ethers were chimyl . SGG is present in relatively small amounts in brain (17). Testicular SGG appears to be present mainly in germinal cells (12). Sulfation of galactosylacylalkylglycerol (GG) to SGG has been shown to be restricted to a briefphase early in spermatogenesis (11,14) and SGG is maintained in subsequent germinal cell stages without apparent turnover (11), despite cell division and extensive membrane reorganization. We have recently reported (18) the preparation in rabbits of I Previously referred to as sulfogalactoglycerolipid .

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Section: Discussionmentioning

confidence: 90%

Section: Cell Purificationmentioning

confidence: 99%

Section: Cell Purificationmentioning

confidence: 99%

See 1 more Smart Citation

Localization of sulfatoxygalactosylacylalkylglycerol at the surface of rat testicular germinal cells by immunocytochemical techniques: pH dependence of a nonimmunological reaction between immunoglobulin and germinal cells.

Lingwood¹,

Schachter²

1981

The Journal of Cell Biology

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“…Subsequently, Letts et al [88] (in Schachter’s group) confirmed that the biosynthesis of SGG occurred in primary spermatocytes. Their experimentation involved injection of Na 2 35 SO 4 into adult rats.…”

Section: Biosynthesis and Presence Of Sgg In Male Reproductive Cellsmentioning

confidence: 99%

Properties, metabolism and roles of sulfogalactosylglycerolipid in male reproduction

Tanphaichitr

Kongmanas

Faull

et al. 2018

Progress in Lipid Research

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Sulfogalactosylglycerolipid (SGG, aka seminolipid) is selectively synthesized in high amounts in mammalian testicular germ cells (TGCs). SGG is an ordered lipid and directly involved in cell adhesion. SGG is indispensable for spermatogenesis, a process that greatly depends on interaction between Sertoli cells and TGCs. Spermatogenesis is disrupted in mice null for Cgt and Cst, encoding two enzymes essential for SGG biosynthesis. Sperm surface SGG also plays roles in fertilization. All of these results indicate the significance of SGG in male reproduction. SGG homeostasis is also important in male fertility. Approximately 50% of TGCs become apoptotic and phagocytosed by Sertoli cells. SGG in apoptotic remnants needs to be degraded by Sertoli lysosomal enzymes to the lipid backbone. Failure in this event leads to a lysosomal storage disorder and sub-functionality of Sertoli cells, including their support for TGC development, and consequently subfertility. Significantly, both biosynthesis and degradation pathways of the galactosylsulfate head group of SGG are the same as those of sulfogalactosylceramide (SGC), a structurally related sulfoglycolipid important for brain functions. If subfertility in males with gene mutations in SGG/SGC metabolism pathways manifests prior to neurological disorder, sperm SGG levels might be used as a reporting/predicting index of the neurological status.

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“…FT has also been implicated in the development of cell-cell adhesions during synaptogenesis of neurons in culture [2] and during mammalian fertilization [29]. FT activity in the mouse testis was first reported by Schachter and co-workers [30,31]. Later, Millette and his associates [3,32] showed that mouse testis contains very high levels of total FT and that FT activity is associated with the germ cell surface.…”

Section: Discussionmentioning

confidence: 98%

Presence of Multiple Fucosyltransferases in Rat Sertoli Cells and Spermatogenic Cells1

Raychoudhury

Millette

1994

Biology of Reproduction

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Differential expression of fucosyltransferases (FTs) on Sertoli cell and germ cell surfaces and their function as ectoenzymes may be important in the process of spermatogenesis. To determine the glycosidic linkage specificity of FTs present in cultured Sertoli cells and in germ cells, we quantified FT activities by thin-layer chromatography using both high and low molecular weight acceptors in the presence of GDP-[14C]-L-fucose. Analysis of the acceptor substrate specificity of the FTs indicated that alpha(1-2), alpha(1-3), alpha(1-4)-FTs are expressed as demonstrated by fucose incorporation into phenyl-beta-D-galactoside, 2'-fucosyllactose, and lacto-N-fucopentaose-I, respectively. In Sertoli cells, the ratios of the three FTs examined were the same for whole-cell extracts and samples of purified plasma membranes. Higher relative FT activity was observed in plasma membranes from mixed germ cells than in Sertoli cell membranes. Furthermore, alpha(1-3)-FT and alpha(1-4)-FT activities were higher in mixed germ cell membranes. Spermatogenic stage specificity of FT expression was assessed in purified populations of germ cells. With calculation on a per-cell basis, all three alpha-FTs exhibited a quantitative decrease during the transition between pachytene spermatocytes and round spermatids. The decrease in alpha(1-3)-FT activity was particularly significant. In rat germ cells, all three alpha-FT activities associated with the cell surface in pachytene spermatocytes and round spermatids were 34-53% and 52-53%, respectively, of the total cell FT activity.(ABSTRACT TRUNCATED AT 250 WORDS)

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Late spermatocytes from immature rat testis isolation, electron microscopy, lectin agglutinability and capacity for glycoprotein and sulfogalactoglycerolipid biosynthesis

Cited by 22 publications

References 27 publications

Localization of sulfatoxygalactosylacylalkylglycerol at the surface of rat testicular germinal cells by immunocytochemical techniques: pH dependence of a nonimmunological reaction between immunoglobulin and germinal cells.

Localization of sulfatoxygalactosylacylalkylglycerol at the surface of rat testicular germinal cells by immunocytochemical techniques: pH dependence of a nonimmunological reaction between immunoglobulin and germinal cells.

Properties, metabolism and roles of sulfogalactosylglycerolipid in male reproduction

Presence of Multiple Fucosyltransferases in Rat Sertoli Cells and Spermatogenic Cells1

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