Histochemical Differentiation of the Basement Membrane of the Mouse Seminiferous Tubule

1962 ◽  
Vol s3-103 (63) ◽  
pp. 385-391
Author(s):  
A. H. BAILLIE
Keyword(s):  
Basement Membrane ◽  
Protein Metabolism ◽  
Seminiferous Tubule ◽  
Glucuronic Acid ◽  
Chondroitin Sulphate ◽  
Hair Follicles ◽  
Ground Substance ◽  
Seminiferous Tubules ◽  
Active Protein ◽  
Pas Reaction

The ground substance of the testis of the albino mouse is PAS-positive but not metachromatic, and probably highly aggregated. The basement of the seminiferous tubules is intensely PAS-positive, metachromatic, and possibly not so highly aggregated. The reactivity of the ground substance to the PAS reaction and toluidine blue is tentatively ascribed to the presence of chondroitin sulphate C: this compound, previously known to contain N acetyl-galactosamine, glucuronic acid, tyrosine and tryptophane, is associated with arginine. The genesis of the basement membrane of the seminiferous tubule is shown to include the formation of a sheath of atypical elongated fibroblasts, the secretion of a PAS positive, metachromatic substance associated with arginine between this sheath and the seminiferous tubule, the appearance of mitochondria in the cells of the sheath, and lastly, the acquisition of alkaline phosphatase by these fibroblasts and its spread to the intervening ground substance. These changes are thought to be related to the structural and nutritional requirements of the seminiferous tubules. In its intense positive reaction to PAS and in its metachromasy, the basement membrane of the seminiferous tubule agrees with the ground substance adjacent to sites of active protein metabolism, such as growing tumours, embryonic organs, hair follicles, and skin.

Examination of basement membrane components associated with the bovine seminiferous tubule basal lamina

2007 ◽  
Vol 19 (3) ◽  
pp. 473 ◽  
Author(s):  
Veronica Glattauer ◽  
Helen F. Irving-Rodgers ◽  
Raymond J. Rodgers ◽  
Sally Stockwell ◽  
Alan G. Brownlee ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Seminiferous Tubule ◽  
Bos Indicus ◽  
Age Groups ◽  
Seminiferous Tubules ◽  
Collagen Type Iv ◽  
Positive Staining ◽  
Type I ◽  
Type Iv ◽  
Lectin Staining

Immunohistology has been used to examine the distribution of certain components of the basement membrane (BM) associated with bovine spermatogonial germ cells that are located within the seminiferous tubules. Histology was performed on testis tissue from Brahman cattle (Bos indicus) of three different age groups: pre-pubescent (4–6 months), juvenile (8–10 months) and adult (18–24 months) animals. There were no major changes in the BM composition apparent between these three age groups, except for certain lectin staining. These data suggest that the predominant collagen type IV component may have an α3 and α4 composition, although other chains, including the α5 and α6 chains, were also present. Possibly the main laminin type present was laminin 121 (α1β2γ1), although other variants were also present. Both nidogen-1 and perlecan, which are normal BM components, were also found as part of the seminiferous tubule BM. Interstitial collagens, such as type I, III and VI collagens, were found in the peritubular space, but were not part of the BM itself, although type VI collagen was most visible in the peritubular zone adjacent to the tubules. Examination of the BM with a range of lectins gave strong staining for (glcNAc)2 entities, weak positive staining for α-l-fuc, but little or no staining for α-galNAc and (glcNAc)3 at all ages, whereas staining for α-gal, β-gal(1→3)galNAc and α-man showed developmental changes.

Ultrastructural Differentiation of the Basement Membrane of the Mouse Seminiferous Tubule

1964 ◽  
Vol s3-105 (70) ◽  
pp. 203-207
Author(s):  
A. H. BAILLIE
Keyword(s):  
Basement Membrane ◽  
Sertoli Cells ◽  
Seminiferous Tubule ◽  
Plasma Membranes ◽  
Fine Particles ◽  
Extracellular Fluid ◽  
Outer Zone ◽  
Seminiferous Tubules ◽  
Postnatal Life ◽  
Sheath Cells

The development of the basement membrane of the mouse seminiferous tubule has been studied between birth and the end of the tenth week of postnatal life by means of the electron microscope. The sheath cells which surround the seminiferous tubules are derived from mesenchymal cells, and differentiation of these specialized cells continues for 5 weeks after birth. Between the plasma membranes of the sheath cells and those of the Sertoli cells there are 3 distinct zones. The zone of decreased electron density nearest the seminiferous tubule is thought to be specialized extracellular fluid filtered by the second or intermediate zone, which consists of numerous fine particles, and the term ‘pseudomembrane’ is advanced for this zone. Collagen develops in the broad outer zone next to the sheath cells and fulfils the support requirements of the growing tubule.

Anionin sites of the basement membrane of rat seminiferous tubules during ontogenesis

1988 ◽  
Vol 63 (1) ◽  
pp. 83-87
Author(s):  
Bruno P. Leheup ◽  
Jean-Louis Gelly ◽  
Jean-Luc Delongeas ◽  
Georges Grignon
Keyword(s):  
Basement Membrane ◽  
Seminiferous Tubules

scholarly journals Meiosis, spermatogenesis and ultrastructure of the basement membrane of seminiferous tubules in patients with azoospermia

2019 ◽  
Vol 20 (1) ◽  
pp. 43-54
Author(s):  
E. E. Bragina ◽  
I. I. Vityazeva ◽  
M. A. Lelekova ◽  
A. A. Kashintsova ◽  
S. V. Bogolyubov ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Seminiferous Tubules

Validation of macroscopic maturity stages of the Patagonian red octopus Enteroctopus megalocyathus

2012 ◽  
Vol 93 (3) ◽  
pp. 833-842 ◽  
Author(s):  
Nicolas Ortiz
Keyword(s):  
Sexual Maturation ◽  
Reproductive System ◽  
Seminiferous Tubule ◽  
Atlantic Coast ◽  
Microscopic Level ◽  
Maturity Index ◽  
Follicular Cell ◽  
Seminiferous Tubules ◽  
Cell Complexes ◽  
Maturation Stages

Testes and ovaries of Enteroctopus megalocyathus collected along the Patagonian Atlantic coast were analysed histologically to validate the macroscopic maturity scales adopted for this species. Changes through the course of development of the seminiferous tubules and of the oocyte/follicular cell complexes were characterized and these were classified into five and six microscopic categories of development respectively. A histological maturity index, based on the frequencies of microscopic categories, was used to assess the correspondence between macroscopic maturation stages and the microscopic level of development of the gonadal tissue. Seminiferous tubules showed a regular and progressive pattern of microscopic development within each macroscopic stage and between consecutive macroscopic stages. However, a minority of males exhibiting seminiferous tubule with sperm did not display macroscopic characteristics of the mature-spawning stage. In females, an overlapping of microscopic categories was observed in maturing macroscopic stages. Previtellogenic oocytes were not present at mature-spawning or spent stages. Significant changes in the histological maturity index were observed between consecutive macroscopic stages, confirming the validity of macroscopic maturity scales of both sexes. In addition, by considering both macroscopic and microscopic criteria, it was possible to determine the overall state of development and functioning of the reproductive system during sexual maturation of this species.

scholarly journals Intra-testicular injection of adenoviral constructs results in Sertoli cell-specific gene expression and disruption of the seminiferous epithelium

Reproduction ◽  
2009 ◽  
Vol 137 (2) ◽  
pp. 361-370 ◽  
Author(s):  
R P Hooley ◽  
M Paterson ◽  
P Brown ◽  
K Kerr ◽  
P T K Saunders
Keyword(s):  
Seminiferous Tubule ◽  
Fluorescent Protein ◽  
Immune Cell ◽  
Adenoviral Vectors ◽  
Seminiferous Tubules ◽  
Specific Gene ◽  
Viral Particles ◽  
Junctional Complexes ◽  
Testicular Injection

Spermatogenesis is a complex process that cannot be modelledin vitro. The somatic Sertoli cells (SCs) within the seminiferous tubules perform a key role in supporting maturation of germ cells (GCs). Progress has been made in determining what aspects of SC function are critical to maintenance of fertility by developing rodent models based on the Cre/LoxP system; however, this is time-consuming and is only applicable to mice. The aim of the present study was to establish methods for direct injection of adenoviral vectors containing shRNA constructs into the testis as a way of inducing target-selective knock-downin vivo. We describe here a series of experiments using adenovirus expressing a green fluorescent protein (GFP) transgene. Injection via the efferent ductules resulted in SC-specific expression of GFP; expression levels paralleled the amount of infective viral particles injected. At the highest doses of virus seminiferous tubule architecture were grossly disturbed and immune cell invasion noted. At lower concentrations, the expression of GFP was variable/negligible, the seminiferous tubule lumen was maintained but stage-dependent GC loss and development of numerous basal vacuoles was observed. These resembled intercellular dilations of SC junctional complexes previously described in rats and may be a consequence of disturbances in SC function due to interaction of the viral particles with the coxsackie/adenovirus receptor that is a component of the junctional complexes within the blood testis barrier. In conclusion, intra-testicular injection of adenoviral vectors disturbs SC functionin vivoand future work will therefore focus on the use of lentiviral delivery systems.

scholarly journals Characterization and Expression of the Laminin γ3 Chain: A Novel, Non-Basement Membrane–associated, Laminin Chain

1999 ◽  
Vol 145 (3) ◽  
pp. 605-618 ◽  
Author(s):  
Manuel Koch ◽  
Pamela F. Olson ◽  
Anne Albus ◽  
William Jin ◽  
Dale D. Hunter ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Basement Membranes ◽  
Chromosome 9 ◽  
Seminiferous Tubules ◽  
Apical Surface ◽  
Gene Encoding ◽  
Ductus Deferens ◽  
Functional Roles ◽  
Cloned Gene ◽  
Ciliated Epithelial Cells

Laminins are heterotrimeric molecules composed of an α, a β, and a γ chain; they have broad functional roles in development and in stabilizing epithelial structures. Here, we identified a novel laminin, composed of known α and β chains but containing a novel γ chain, γ3. We have cloned gene encoding this chain, LAMC3, which maps to chromosome 9 at q31-34. Protein and cDNA analyses demonstrate that γ3 contains all the expected domains of a γ chain, including two consensus glycosylation sites and a putative nidogen-binding site. This suggests that γ3-containing laminins are likely to exist in a stable matrix. Studies of the tissue distribution of γ3 chain show that it is broadly expressed in: skin, heart, lung, and the reproductive tracts. In skin, γ3 protein is seen within the basement membrane of the dermal-epidermal junction at points of nerve penetration. The γ3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules. The distribution of γ3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes. It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

scholarly journals Sequential degradation of a chondroitin sulphate trisaccharide by lysosomal enzymes from embryonic-chick epiphysial cartilage

1979 ◽  
Vol 179 (1) ◽  
pp. 7-13 ◽  
Author(s):  
B Ingmar ◽  
A Wasteson
Keyword(s):  
Lysosomal Enzymes ◽  
Glucuronic Acid ◽  
Enzyme Inhibitors ◽  
Functional Unit ◽  
Chondroitin Sulphate ◽  
Cartilage Degradation ◽  
Ester Group ◽  
Embryonic Chick ◽  
Reaction Products ◽  
Epiphysial Cartilage

The disulphated trisaccharide D-N-acetylgalactosamine sulphate-beta-D-glucuronic acid-beta-D-N-acetylgalactosamine sulphate prepared from 35S- or 14C-labelled chondroitin sulphate was incubated with a preparation of lysosomal enzymes from embryonic-chick epiphysial cartilage. Degradation was demonstrated by analysis of the reaction products. By use of the appropriate intermediate products as substrates, in conjunction with specific enzyme inhibitors, it was shown that the degradation proceeded sequentially from the non-reducing end. It was initiated by sulphatase (preferentially hydrolysing sulphate ester groups at the 6-position), followed by beta-N-acetylgalactosaminidase and beta-glucuronidase, converting the substrate into monosaccharides and inorganic sulphate. The latter enzyme preferentially attacked disaccharides carrying their sulphate ester group at C-4 of the hexosamine residue. Generation of chondroitin sulphate oligosaccharides may occur by the action of an endoglycosidase, previously demonstrated in embryonic-chick cartilage. Endo- and exo-enzymes may thus form a functional unit in lysosomal degradation of chondroitin sulphate.

286 EXPRESSION OF PLURIPOTENT STEM CELL MARKERS IN DOMESTIC CAT SPERMATOGONIAL CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 290 ◽  
Author(s):  
R. H. Powell ◽  
M. N. Biancardi ◽  
J. Galiguis ◽  
Q. Qin ◽  
C. E. Pope ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Basement Membrane ◽  
Germ Cell ◽  
Germ Cells ◽  
Spermatogonial Stem Cells ◽  
Seminiferous Tubules ◽  
Cell Populations ◽  
Surface Markers ◽  
Cell Markers

Spermatogonial stem cells (SSC), progenitor cells capable of both self-renewal and producing daughter cells that will differentiate into sperm, can be manipulated for transplantation to propagate genetically important males. This application was demonstrated in felids by the successful xeno-transplantation of ocelot mixed germ cells into the testes of domestic cats, which resulted in the production of ocelot sperm (Silva et al. 2012 J. Androl. 33, 264–276). Spermatogonial stem cells are in low numbers in the testis, but have been identified and isolated in different mammalian species using SSC surface markers; however, their expression varies among species. Until recently, little was known about the expression of SSC surface markers in feline species. We previously demonstrated that many mixed germ cells collected from adult cat testes express the germ cell markers GFRα1, GPR125, and C-Kit, and a smaller population of cells expresses the pluripotent SSC-specific markers SSEA-1 and SSEA-4 (Powell et al. 2011 Reprod. Fertil. Dev. 24, 221–222). In the present study, our goal was to identify germ cell and SSC-specific markers in SSC from cat testes. Immunohistochemical (IHC) localization of germ cell markers GFRα1, GPR125, and C-Kit and pluripotent SSC-specific markers SSEA-1, SSEA-4, TRA-1-60, TRA-1-81, and Oct-4 was detected in testis tissue from both sexually mature and prepubertal males. Testes were fixed with modified Davidson’s fixative for 24 h before processing, embedding, and sectioning. The EXPOSE Mouse and Rabbit Specific HRP/DAB detection IHC kit (Abcam®, Cambridge, MA, USA) was used for antibody detection. Staining for SSEA-1, SSEA-4, TRA-1-60, TRA-1-81, and Oct-4 markers was expressed specifically at the basement membrane of the seminiferous tubules in both adult and prepubertal testes. The GFRα1 and GPR125 markers were detected at the basement membrane of the seminiferous tubules and across the seminiferous tubule section. However, C-Kit was not detected in any cell. Using flow cytometry from a pool of cells from seven adult testes, we detected 45% GFRα1, 50% GPR125, 59% C-Kit, 18% TRA-1-60, 16% TRA-1-81 positive cells, and a very small portion of SSEA-1 (7%) and SSEA-4 (3%) positive cells. Dual staining of germ cells pooled from 3 testes revealed 3 distinct cell populations that were positive for GFRα1 only (23%), positive for both GFRα1 and SSEA-4 (6%), and positive for SSEA-4 only (1%). Our IHC staining of cat testes indicated that cells along the basement membrane of seminiferous tubules were positive for SSC-specific markers, and flow cytometry analysis revealed that there were different cell populations expressing both germ cell and SSC-specific markers. Flow cytometry results show overlapping germ cell populations expressing SSEA-4 and GFRα1, and IHC results reveal that SSEA-4 positive cells are spermatogonia, whereas GFRα1 positive cells include other stages of germ cells, indicating that the small population of cells positive only for SSEA-4 is undifferentiated cat SSC.

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