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).

Alteration of nuclear architecture in male germ cells of chromosomally derived subfertile mice

2001 ◽  
Vol 114 (24) ◽  
pp. 4429-4434
Author(s):  
Silvia Garagna ◽  
Maurizio Zuccotti ◽  
Alan Thornhill ◽  
Raul Fernandez-Donoso ◽  
Soledad Berrios ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Spatial Organization ◽  
Nuclear Architecture ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Condensed State ◽  
Germ Cell Differentiation ◽  
Y Chromosomes ◽  
Dual Colour ◽  
Pachytene Spermatocytes

The mammalian cell nucleus consists of numerous compartments involved in the regular unfolding of processes such as DNA replication and transcription, RNA maturation, protein synthesis and cell division. Knowledge is increasing of the relationships between high-order levels of chromatin organization and its spatial organization, and of how these relationships contribute to the various functions carried out in the nucleus. We have studied the spatial arrangement of mouse telocentric chromosomes 5, 11, 13, 15, 16 and 17, some of their metacentric Robertsonian derivatives, and X and Y chromosomes by whole chromosome painting in male germ (spermatogonia, pachytene spermatocytes and spermatids) and Sertoli cells of homozygous and heterozygous individuals. Using dual-colour fluorescence in situ hybridization we found that these chromosomes occupy specific nuclear territories in each cell type analysed. When chromosomes are present as Robertsonian metacentrics in the heterozygous state, that is, as Robertsonian metacentrics and their homologous telocentrics, differences in their nuclear positions are detectable: heterozygosity regularly produces a change in the nuclear position of one of the two homologous telocentrics in all the cell types studied. In the Robertsonian heterozygotes, the vast majority of the Sertoli cells show the sex chromosomes in a condensed state, whereas they appear decondensed in the Robertsonian homozygotes. As the Robertsonian heterozygosities we studied produce a chromosomally derived impairment of male germ-cell differentiation, we discuss the possibility that changes in chromosome spatial territories may alter some nuclear machinery (e.g., synapsis, differential gene expression) important for the correct unfolding of the meiotic process and for the proper functioning of Sertoli cells.

Studies of Sl/Sld in equilibrium with +/+ mouse aggregation chimaeras. II. Effect of the steel locus on spermatogenesis

Development ◽  
1988 ◽  
Vol 102 (1) ◽  
pp. 117-126 ◽  
Author(s):  
H. Nakayama ◽  
H. Kuroda ◽  
H. Onoue ◽  
J. Fujita ◽  
Y. Nishimune ◽  
...  
Keyword(s):  
Mast Cells ◽  
Germ Cell ◽  
Germ Cells ◽  
Cross Sections ◽  
Sertoli Cells ◽  
Type A ◽  
Precursor Cells ◽  
Intrinsic Defect ◽  
Serial Sections ◽  
Type A Spermatogonia

Mutant mice of Sl/Sld genotype are deficient in melanocytes, erythrocytes, mast cells and germ cells. Deficiency of melanocytes, erythrocytes and mast cells is not attributable to an intrinsic defect in their precursor cells but to a defect in the tissue environment that is necessary for migration, proliferation and/or differentiation. We investigated the mechanism of germ cell deficiency in male Sl/Sld mice by producing aggregation chimaeras from Sl/Sld and +/+ embryos. Chimaeric mice with apparent white stripes were obtained. Two of four such chimaeras were fertile and the phenotypes of resulting progenies showed that some Sl/Sld germ cells had differentiated into functioning sperms in the testis of the chimaeras. In cross sections of the testes of chimaeras, both differentiated and nondifferentiated tubules were observed. However, the proportions of type A spermatogonia to Sertoli cells in both types of tubules were comparable to the values observed in differentiated tubules of normal +/+ mice. We reconstructed the whole length of four tubules from serial sections. Differentiated and nondifferentiated segments alternated in a single tubule. The shortest differentiated segment contained about 180 Sertoli cells and the shortest nondifferentiated segment about 150 Sertoli cells. These results suggest that Sertoli cells of either Sl/Sld or +/+ genotype make discrete patches and that differentiation of type A spermatogonia does not occur in patches of Sl/Sld Sertoli cells.

Fertilization and embryonic developmental capacity of epididymal and testicular sperm and immature spermatids and spermatocytes

1997 ◽  
Vol 6 (1) ◽  
pp. 55-68 ◽  
Author(s):  
Orly Lacham-Kaplan ◽  
Alan Trounson
Keyword(s):  
Close Contact ◽  
Mammalian Species ◽  
Type B ◽  
Final Phase ◽  
Testicular Sperm ◽  
Primitive Type ◽  
Developmental Capacity ◽  
Form Type ◽  
Mitotic Proliferation ◽  
Type A Spermatogonia

Spermatogenesis in mammalian species begins after birth. The gonocytes, arrested at G2 of the cell cycle in the foetus, resume mitotic proliferation after birth. As identified in the mouse, the gonocytes migrate towards the periphery of the seminiferous cords at day 4 to day 6 after birth and are located in close contact with the basal lamina. From this stage the gonocytes are referred to as primitive type A spermatogonia. These cells continue mitotic proliferation and differentiate to form type B spermatogonia. By day 10 after birth, many of the type B spermatogonia have formed preleptotene primary spermatocytes which undergo a final phase of DNA synthesis (leptotene) prior to entering meiotic prophase (zygotene).

scholarly journals Cloning, pharmacological characterization, and expression analysis of Atlantic salmon (Salmo salar L.) nuclear progesterone receptor

Reproduction ◽  
2011 ◽  
Vol 141 (4) ◽  
pp. 491-500 ◽  
Author(s):  
Shi X Chen ◽  
Jan Bogerd ◽  
Eva Andersson ◽  
Fernanda F L Almeida ◽  
Geir Lasse Taranger ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Atlantic Salmon ◽  
Sertoli Cells ◽  
Plasma Levels ◽  
Type A ◽  
Mrna Levels ◽  
Early Type ◽  
Type B ◽  
Late Type ◽  
Nuclear Progesterone Receptor

To better understand the role(s) of progestogens during early stages of spermatogenesis, we carried out studies on the nuclear progesterone receptor (Pgr) of the Atlantic salmon. Its open-reading frame shows the highest similarity with other piscine Pgr proteins. When expressed in mammalian cells, salmon Pgr exhibited progestogen-specific, dose-dependent induction of reporter gene expression, with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) showing the highest potency. We then analyzed testicular pgr mRNA and DHP plasma levels in animals during the onset of spermatogenesis, which were exposed to natural light or to constant light, to induce significant differences in testis growth. Grouping of the animals according to their progress through spermatogenesis showed that testicular pgr mRNA levels as well as DHP plasma levels first increased when germ cells had reached the stage of late type B spermatogonia and further increased when entered meiosis, i.e. when spermatocytes were present. However, in situ hybridization studies revealed that pgr mRNA expression was restricted to Sertoli cells, with a strong signal in Sertoli cells contacting type A/early type B spermatogonia, while Sertoli cells contacting larger germ cell clones with further differentiated stages (e.g. late type B spermatogonia) were less intensely/not stained. We conclude that the increase in pgr mRNA levels per pair of testis reflects, at least in part, the increased number of Sertoli cells enveloping type A and early type B spermatogonia. We propose that Sertoli cell-expressed Pgr may mediate DHP-stimulated early steps in spermatogenesis in Atlantic salmon, such as an increase in the number of new spermatogonial cysts.

scholarly journals Non-random distribution of spermatogonia in rats: evidence of niches in the seminiferous tubules

Reproduction ◽  
2003 ◽  
pp. 669-680 ◽  
Author(s):  
H Chiarini-Garcia ◽  
AM Raymer ◽  
LD Russell
Keyword(s):  
Basal Lamina ◽  
Random Distribution ◽  
Type A ◽  
Seminiferous Epithelium ◽  
Seminiferous Tubules ◽  
Interstitial Tissue ◽  
Primitive Type ◽  
Linear Index ◽  
Type A Spermatogonia

The relationships and distribution of spermatogonia were studied as a function of the stage of the seminiferous epithelium cycle in rats. Primitive spermatogonia in the mouse are located along regions of the basal lamina that face the interstitium. Before studying the distribution of spermatogonia in rats, it was necessary to characterize the various types of spermatogonia, as recently performed for mice. The Strauss' linear index (Li) selectivity method was then used and spermatogonia of the A(single) (A(s)) to A(aligned) (A(al)) lineage were preferentially found to be located in regions opposing the interstitium at stages V, VII and IX of the spermatogenic cycle. Because relatively little tubule-to-tubule contact occurs in rats, the aim of this study was to determine whether tubule-to-tubule contact or tubule proximity (or alternatively, the amount of interstitium) was an important factor in spermatogonial position. In this regard, another method (tubule proximity) was devised to determine spermatogonial position that accounted for the presence of adjacent tubules. This method showed that the position of tubules, rather than tubule contact, was more accurate than the Li method in determining the location of spermatogonia in the rat. The results also showed a non-random distribution of spermatogonia resembling that of the mouse, and that tubule-to-tubule contact is not essential for the positioning of spermatogonia. In conclusion, the results of this study strongly indicate that the most primitive type A spermatogonia (A(s), A(paired) and A(al)) in rats are present in niches located in those areas of the seminiferous tubules that border the interstitial tissue.

scholarly journals DNA analysis and sorting of viable mouse testis cells.

1981 ◽  
Vol 29 (6) ◽  
pp. 738-746 ◽  
Author(s):  
W M Grogan ◽  
W F Farnham ◽  
J M Sabau
Keyword(s):  
Dna Content ◽  
Adult Mouse ◽  
Dna Analysis ◽  
Cell Types ◽  
Mouse Testis ◽  
Spermatogenic Cells ◽  
Hoechst 33342 ◽  
Cell Sorter ◽  
Pachytene Spermatocytes

The dye Hoechst 33342 and a 2-parameter cell sorter have been used to measure DNA content in viable testis cells and to sort pachytene spermatocytes and round spermatids from adult mouse testis to virtually 100% homogeneity. Early diploid spermatogenic cells were enriched to 90%, a 10-fold purification. The capability for viable sorting of most testis cell types to homogeneity in numbers suitable for many biochemical applications is demonstrated.

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.

Evidence for the control of testicular interstitial fluid volume in the rat by specific germ cell types

1991 ◽  
Vol 128 (3) ◽  
pp. 359-NP ◽  
Author(s):  
R. M. Sharpe ◽  
J. M. S. Bartlett ◽  
G. Allenby
Keyword(s):  
Germ Cell ◽  
Sertoli Cells ◽  
Interstitial Fluid ◽  
Local Heating ◽  
Cell Types ◽  
Volume Changes ◽  
Interstitial Fluid Volume ◽  
Testicular Weight ◽  
Pachytene Spermatocytes

ABSTRACT Following on from our recent evidence that Sertoli cells may regulate testicular interstitial fluid (IF) volume, this study has assessed whether depletion of specific germ cell types in vivo is associated with changes in recovered IF volume. Germ cell depletion was induced by either a single oral administration of 650 mg methoxyacetic acid (MAA)/kg or exposure of the testes to local heating (43 °C for 30 min). Treatment with MAA induced depletion or loss of most pachytene and later spermatocytes at 1–3 days and, because of maturation depletion, this resulted in the specific depletion of later germ cell types at 7–35 days. Testicular IF volume was unchanged at 1–7 days after MAA treatment but was increased significantly (P < 0·01) at 14 days and was nearly doubled (P< 0·001) at 21 days, before returning to control levels at 28–42 days. Serum LH (and FSH) levels were generally higher in MAA-treated rats, especially at 21 and 28 days, but there was no obvious correlation between LH levels and IF volume changes. Similarly, there was no relationship between IF volume changes and testicular weight or IF levels of testosterone. The increase in IF volume at 14–21 days after MAA treatment coincided with specific depletion of the later elongate spermatids (steps 14–19) and, when these cells reappeared in the testis, IF volume normalized. This possible causal association was studied further in rats exposed to local testicular heating which, within 3 days, caused major depletion of pachytene spermatocytes and early (step 1–8) spermatids. However, testicular IF volume in heat-exposed rats did not change until 14 days, a time at which depletion of the later (step 9–19) spermatids first became evident; IF volume remained increased whilst these germ cells were absent or depleted. The pattern of change in IF volume in heat-exposed rats was not related to LH (or FSH) levels, which were raised at most time-points after heat treatment, nor to testicular weight which was decreased considerably at 3 days and declined progressively thereafter. These data thus provide evidence that specific depletion of the most mature germ cell types (the elongate spermatids) is associated with specific changes in testicular IF volume, presumably via modulation of the secretion of vasoactive factors by the Sertoli cells. These findings also reinforce the growing evidence for the mutual interdependence of all of the cell types in the testis. Journal of Endocrinology (1991) 128, 359–367

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