scholarly journals PRKRA Localizes to Nuage Structures and the Ectoplasmic Specialization and Tubulobulbar Complexes in Rat and Mouse Testis

2014 ◽  
Vol 2014 ◽  
pp. 1-9
Author(s):  
Junya Suzuki ◽  
Sadaki Yokota
Keyword(s):  
Sertoli Cells ◽  
Rna Silencing ◽  
Binding Proteins ◽  
Spermatogenic Cells ◽  
P Bodies ◽  
Cytoplasmic Rna ◽  
Dsrna Binding ◽  
Chromatoid Bodies ◽  
Pachytene Spermatocytes ◽  
Ectoplasmic Specialization

The cytoplasmic RNA-induced silencing complex (RISC) contains dsRNA binding proteins, including PRKRA, TRBP, and Dicer. RISC localizes to P-bodies. The nuage of the spermatogenic cells has function similar to the P-bodies. We study whether PRKRA localizes to nuage of spermatogenic cells of rat and mouse. PRKRA localized to four types of nuage structures, including aggregates of 60–90 nm particles, irregularly-shaped perinuclear granules, and intermitochondrial cement of pachytene spermatocytes, and chromatoid bodies of round spermatids. In addition, PRKRA is associated with dense material surrounding tubulobulbar complexes and with the ectoplasmic specialization. The results suggest that PRKRA functions in the nuage as an element of RNA silencing system and plays unknown role in the ectoplasmic specialization and at the tubulobulbar complexes of Sertoli cells attaching the head of late spermatids.

scholarly journals Pachytene spermatocyte and round spermatid binding to Sertoli cells in vitro

1988 ◽  
Vol 90 (1) ◽  
pp. 105-114
Author(s):  
G.C. Enders ◽  
C.F. Millette
Keyword(s):  
Sertoli Cell ◽  
Sertoli Cells ◽  
Round Spermatid ◽  
Pachytene Spermatocyte ◽  
Assay System ◽  
Spermatogenic Cells ◽  
Vitro Assay ◽  
Cell Monolayers ◽  
Pachytene Spermatocytes

Spermatogenic cells differentiate in vivo while in continuous contact with the Sertoli cell. During differentiation, the spermatogenic cells and Sertoli cells form a number of morphologically distinct stage-specific adhesions. We describe an in vitro assay system for studying the adhesion of spermatogenic cells to Sertoli cell monolayers. Mixed populations of spermatogenic cells or enriched fractions of pachytene spermatocytes and round spermatids were labelled with the vital dye, fluorescein diacetate, prior to their addition to Sertoli cell monolayers so that the adhesion of viable spermatogenic cells could be quantified. Using this assay system, the number of pachytene spermatocyte and round spermatid binding sites on the Sertoli cell monolayer were similar, but the kinetics of binding were different. Pachytene spermatocytes were able to inhibit significantly round spermatid binding, while round spermatids did not significantly inhibit pachytene spermatocyte binding. After coculture for 24–48 h, spermatocytes form junctional structures with Sertoli cells that are similar to desmosome-like junctions. These results suggest that pachytene spermatocytes and round spermatids bind to Sertoli cells by different mechanisms.

scholarly journals Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization

1977 ◽  
Vol 74 (1) ◽  
pp. 68-85 ◽  
Author(s):  
AR Bellve ◽  
JC Cavicchia ◽  
CF Millette ◽  
DA O'Brien ◽  
YM Bhatnagar ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Morphological Characteristics ◽  
Type A ◽  
Cell Types ◽  
Specific Cell ◽  
Spermatogenic Cells ◽  
Type B ◽  
Primitive Type ◽  
Pachytene Spermatocytes ◽  
Type A Spermatogonia

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).

scholarly journals The effect of testosterone, dihydrotestosterone and oestradiol on the re-initiation of spermatogenesis in the adult photoinhibited Djungarian hamster

2007 ◽  
Vol 192 (3) ◽  
pp. 553-561 ◽  
Author(s):  
Sarah J Meachem ◽  
Stefan Schlatt ◽  
Saleela M Ruwanpura ◽  
Peter G Stanton
Keyword(s):  
Sertoli Cells ◽  
Cell Number ◽  
Steroid Treatment ◽  
Spermatogenic Cells ◽  
Type B ◽  
Djungarian Hamster ◽  
Cell Numbers ◽  
Optical Disector ◽  
Pachytene Spermatocytes ◽  
Type A Spermatogonia

The roles of testosterone (T) and its metabolites on hamster spermatogenesis are poorly defined. This study assessed the effects of T, dihydrotestosterone (DHT) and oestradiol (E) on the re-initiation of spermatogenesis in the adult Djungarian hamster. Hamsters raised under long photoperiods (LD, 16 h light:8 h darkness) were exposed to short photoperiods (SD, 8 h light:16 h darkness) for 11 weeks to suppress gonadotrophins. Groups of eight animals then received T, DHT and E for 5 weeks. Cell numbers were determined using the optical disector (sic). The number of Sertoli cells was suppressed in SD controls to 48% (P < 0.001) of LD control and restored either fully or partially by exogenous DHTand E (2.6- and 1.8-fold above SD levels) respectively, corresponding with a twofold elevation of serum FSH. The number of germ cells in SD animals was reduced (all P < 0.001) to levels reported. The number of type A spermatogonia increased in line with the rise in Sertoli cell number, by 2.6-fold (P < 0.01) and 1.8-fold (NS) above SD controls after DHT and E treatments respectively. DHT increased the number of type B spermatogonia/preleptotene spermatocytes, leptotene/zygotene and pachytene spermatocytes by 3.5-, 5.7- and 21-fold above SD (all P < 0.01) respectively, compared with a 2.2-fold (P < 0.01), 2.4-fold (not significant, NS) and 6-fold (NS) in E-treated animals respectively. Exogenous T had little effect on cell numbers or serum FSH compared with SD controls. Spermatids were rarely observed after steroid treatment. We believe this study suggests that steroids can regulate the re-initiation of early spermatogenic cells via a mechanism which includes FSH.

Annulate lamellae in the Sertoli cells of guinea pig testis after unilateral torsion of the spermatic cord

1984 ◽  
Vol 42 ◽  
pp. 196-197
Author(s):  
J. Chakraborty ◽  
A. P. Sinha Hikim ◽  
J. S. Jhunjhunwala
Keyword(s):  
Sertoli Cells ◽  
Guinea Pigs ◽  
Spermatic Cord ◽  
Present Report ◽  
Cell Types ◽  
Annulate Lamellae ◽  
Spermatogenic Cells ◽  
Electron Microscopic ◽  
Structural Details ◽  
First Time

Although the presence of annulate lamellae was noted in many cell types, including the rat spermatogenic cells, this structure was never reported in the Sertoli cells of any rodent species. The present report is based on a part of our project on the effect of torsion of the spermatic cord to the contralateral testis. This paper describes for the first time, the fine structural details of the annulate lamellae in the Sertoli cells of damaged testis from guinea pigs.One side of the spermatic cord of each of six Hartly strain adult guinea pigs was surgically twisted (540°) under pentobarbital anesthesia (1). Four months after induction of torsion, animals were sacrificed, testes were excised and processed for the light and electron microscopic investigations. In the damaged testis, the majority of seminiferous tubule contained a layer of Sertoli cells with occasional spermatogonia (Fig. 1). Nuclei of these Sertoli cells were highly pleomorphic and contained small chromatinic clumps adjacent to the inner aspect of the nuclear envelope (Fig. 2).

Specific interactions between Dicer-like proteins and HYL1/DRB- family dsRNA-binding proteins in Arabidopsis thaliana

2005 ◽  
Vol 57 (2) ◽  
pp. 173-188 ◽  
Author(s):  
Akihiro Hiraguri ◽  
Riku Itoh ◽  
Naoko Kondo ◽  
Yasuko Nomura ◽  
Daisuke Aizawa ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Binding Proteins ◽  
Specific Interactions ◽  
Dsrna Binding

scholarly journals Dynamics of intracellular calcium induced by lactate and glucose in rat pachytene spermatocytes and round spermatids

Reproduction ◽  
2002 ◽  
pp. 701-710 ◽  
Author(s):  
JG Reyes ◽  
E Herrera ◽  
L Lobos ◽  
K Salas ◽  
N Lagos ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
Carbohydrate Metabolism ◽  
Adenine Nucleotides ◽  
Differential Modulation ◽  
Glycolytic Metabolism ◽  
Spermatogenic Cells ◽  
Lactate Metabolism ◽  
The Media ◽  
Hexose Phosphorylation ◽  
Pachytene Spermatocytes

Glycolytic metabolism in meiotic and post-meiotic spermatogenic cells shows differentiation-related changes. The developmental and physiological significance of these metabolic changes is not known. The aim of the present study was to test the hypothesis that glucose and lactate metabolism can modulate intracellular calcium [Ca2+](i) in spermatogenic cells in an opposing and dynamic manner. Fluorescent probes were used to measure [Ca2+](i) and pH(i), and HPLC was used to measure intracellular adenine nucleotides and mitochondrial sensing of ATP turnover. [Ca2+](i) in pachytene spermatocytes and round spermatids was modulated by changes in lactate and glucose concentrations in the media. The kinetics and magnitude of the [Ca2+](i) changes induced by lactate and glucose were different in meiotic and post-meiotic spermatogenic cells. The presence of glucose in the medium induced a decrease in pH(i) in spermatogenic cells. This glucose-induced pH(i) decrease occurred later than the changes in [Ca2+](i), which were also observed when the pH(i) decrease was inhibited, indicating that the glucose-induced [Ca2+](i) increase was not a consequence of pH(i) changes. Hexose phosphorylation in glycolysis was part of the mechanism by which glucose metabolism induced a [Ca2+](i) increase in spermatogenic cells. The sensitivity of [Ca2+](i) to carbohydrate metabolism was higher in round spermatids than in pachytene spermatocytes. Thus, differentiation-related changes in carbohydrate metabolism in spermatogenic cells determine a dynamic and differential modulation of their [Ca2+](i) by glucose and lactate, two substrates secreted by the Sertoli cells.

Protein synthesis by isolated pachytene spermatocytes in the absence of sertoli cells

1985 ◽  
Vol 233 (2) ◽  
pp. 285-290 ◽  
Author(s):  
N. H. P. M. Jutte ◽  
R. Jansen ◽  
J. A. Grootegoed ◽  
F. F. G. Rommerts ◽  
H. J. van der Molen
Keyword(s):  
Protein Synthesis ◽  
Sertoli Cells ◽  
Pachytene Spermatocytes

scholarly journals Culture patterns and sorting of rat Sertoli cell secretory proteins

1988 ◽  
Vol 89 (2) ◽  
pp. 175-188
Author(s):  
H. Ueda ◽  
L.L. Tres ◽  
A.L. Kierszenbaum
Keyword(s):  
Epithelial Cells ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Secretory Proteins ◽  
Spermatogenic Cells ◽  
Squamous Cells ◽  
Nylon Mesh ◽  
Continuous Sheet ◽  
Peritubular Cells ◽  
Cells Cultured

A cocultivation chamber and two types of permeable substrates have been used to study: (1) the culture patterns of rat Sertoli and peritubular cells, and Sertoli cells cocultured with spermatogenic cells or peritubular cells; and (2) the polarized secretion of Sertoli cell-specific proteins transferrin, S70 and S45-S35 heterodimeric protein. Substrates included a nylon mesh (with openings of 100 micron) coated with extracellular matrix (ECM) material and an uncoated microporous filter (with pores of 0.45 micron). Sertoli cells cultured on ECM-coated nylon mesh organized a continuous sheet of multilayered epithelial cells essentially devoid of spermatogenic cells while peritubular cells formed a layer of squamous cells. Sertoli cells cultured on uncoated microporous substrate formed a continuous sheet of cuboidal epithelial cells with numerous basal cytoplasmic processes projecting into the substrate and abundant apically located spermatogenic cells, while peritubular cells organized one or two layers of loose squamous cells. [35S]methionine-labelled secretory proteins resolved by two-dimensional polyacrylamide gel electrophoresis and autoradiography displayed cell-specific patterns that were slightly influenced by the type of substrate. Sertoli cells cocultured with peritubular cells on uncoated microporous substrate under conditions that enabled separation of apical and basal surfaces, secreted proteins in a polarized fashion. While transferrin was released bidirectionally, S45-S35 heterodimeric protein was released apically. S70 was detected in both apical and basal compartments. We conclude from these studies that: (1) the number of spermatogenic cells decreases when Sertoli-spermatogenic cell cocultures are prepared on ECM-coated nylon substrate; and (2) Sertoli cells in coculture with spermatogenic or peritubular cells on uncoated microporous substrate, organize continuous sheets displaying polarized protein secretion.

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