scholarly journals Isolation and partial characterization of the outer dense fibres and fibrous sheath from the sperm tail of a marsupial: the brushtail possum (Trichosurus vulpecula)

Reproduction ◽  
2001 ◽  
pp. 373-388 ◽  
Author(s):  
M Ricci ◽  
WG Breed
Keyword(s):  
Sucrose Density Gradient ◽  
Blue Staining ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Fibrous Sheath ◽  
Sperm Tail ◽  
Coomassie Blue Staining ◽  
Fibre Network ◽  
Molecular Masses ◽  
Marsupial Species

The flagellum of a mammalian spermatozoon consists of a central axoneme surrounded by two cytoskeletal structures, the outer dense fibres and the fibrous sheath, which may aid in sperm motility or stability. In this study the outer dense fibres and fibrous sheath were isolated and partially characterized in a marsupial species, the brushtail possum (Trichosurus vulpecula). Spermatozoa from the cauda epididymidis were decapitated by sonication, and the head and tail fractions were separated by centrifugation over a 20, 40 and 60% (w/v) sucrose density gradient. After confirming sperm tail purity by Nomarski microscopy, the tails were incubated in either SDS-dithiothreitol to isolate the outer dense fibres or urea-dithiothreitol to isolate the fibrous sheaths. Purified outer dense fibres and fibrous sheaths were solubilized in SDS and beta-mercaptoethanol and proteins were separated by one-dimensional PAGE. Coomassie blue staining showed that the outer dense fibres were composed of seven major proteins (molecular masses: 73, 58, 55, 54, 52, 41 and 16 kDa), and the fibrous sheath was composed of 12 major proteins (molecular masses: 106, 76, 66, 62, 55, 53, 52, 46, 40, 30, 28 and 16 kDa). A polyclonal antibody to the fibrous sheath proteins showed strong crossreactivity with those of fibrous sheath from spermatozoa of several other marsupial species, as well as those from laboratory rats. Subsequent western blotting identified the immunoreactive 76 and 62 kDa proteins from all species, thus indicating their high conservation between species. No crossreactivity of the fibrous sheath antibody to any other cytoskeletal structures, including the outer dense fibres, mid-piece fibre network or connecting laminae, or to the acrosome or underlying subacrosomal material, was evident, indicating that the fibrous sheath proteins are localized to this structure alone. Further work is in progress to determine the extent of homology of these proteins to those in eutherian mammals.

Ontogeny and pathway of formation of 5α-androstane-3α,17β-diol in the testes of the immature brushtail possum Trichosurus vulpecula

2005 ◽  
Vol 17 (6) ◽  
pp. 603 ◽  
Author(s):  
Jean D. Wilson ◽  
Geoffrey Shaw ◽  
Marilyn B. Renfree ◽  
Richard J. Auchus ◽  
Michael W. Leihy ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Tammar Wallaby ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Mature Adults ◽  
Immature Male ◽  
Marsupial Species ◽  
Age Range

The testicular androgen 5α-androstane-3α,17β-diol (androstanediol) mediates virilisation in pouch young of a marsupial, the tammar wallaby, and is the principal androgen formed in immature rodent testes. To chart the pattern of androstanediol formation in another marsupial species, the testes or fragments of testes from brushtail possums (Trichosurus vulpecula) that spanned the age range from early pouch young to mature adults were incubated with 3H-progesterone and the products were identified by high-performance liquid chromatography. The only 19-carbon steroids identified in pouch young and adult testes were the Δ4-3-keto-steroids testosterone and androstenedione. However, androstanediol and another 5α-reduced androgen (androsterone) were synthesised by testes from Day 87–200 males and these appeared to be formed from the 5α-reduction and 3-keto reduction of testosterone and androstenedione. In the prostate and glans penis of the immature male, 3H-androstanediol was converted to dihydrotestosterone. We conclude that the timing of androstanediol formation in the possum testis resembles the process in rodents rather than in the tammar wallaby and that any androstanediol in the circulation probably acts in target tissues via conversion to dihydrotestosterone.

In vitro culture of brushtail possum (Trichosurus vulpecula) epididymal epithelium and induction of epididymal sperm maturation in co-culture

Reproduction ◽  
2000 ◽  
pp. 1-14 ◽  
Author(s):  
M Lin ◽  
X Zhang ◽  
R Murdoch ◽  
RJ Aitken
Keyword(s):  
Sperm Maturation ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Epididymal Sperm ◽  
Cell Monolayers ◽  
Progressive Motility ◽  
Cell Culture Systems ◽  
Marsupial Species ◽  
Morphological Maturation

A medium modified from eutherian systems was used to culture epididymal epithelial cells of the brushtail possum (Trichosurus vulpecula) for more than 2 months. Epididymal tubule fragments from the caput, corpus and cauda epididymides were used to generate cell monolayers. All three epididymal cell culture systems supported maturational changes in marsupial spermatozoa and enabled immature possum spermatozoa to differentiate from a T-shape to a streamlined shape, accompanied by the development of progressive motility after co-culture with 7-day-old cultured epididymal cell monolayers. This epididymal cell and sperm co-culture system for marsupial species may facilitate the identification of specific epithelial factors that affect sperm maturation, particularly in a species in which morphological maturation is readily visible.

Cycle of the seminiferous epithelium in a marsupial species, the brushtail possum (Trichosurus vulpecula), and estimation of its duration

2004 ◽  
Vol 16 (3) ◽  
pp. 307 ◽  
Author(s):  
Minjie Lin ◽  
Amanda Harman ◽  
Terry P. Fletcher
Keyword(s):  
Germ Cells ◽  
Cross Sections ◽  
Seminiferous Epithelium ◽  
Seminiferous Tubules ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Type B ◽  
Semithin Sections ◽  
Marsupial Species ◽  
Kinetics Of

We investigated the cycle of the seminiferous epithelium in a marsupial, namely the brushtail possum (Trichosurus vulpecula), using semithin sections of seminiferous tubules embedded in Spurr’s resin. Using 14 steps of spermatid development as markers, we were able to class tubular cross-sections into 10 well-defined stages of the seminiferous epithelial cycle. The duration of one cycle was 13.5 days, as determined by injections of [3H]-thymidine and autoradiographic examination of the most advanced sperm cells at 2 h and 17 days after injection. The durations of stages I–X were 21.4, 66.4, 54.1, 47.0, 29.8, 28.5, 25.3, 25.0, 12.0 and 15.9 h, respectively, estimated by the relative percentage of occurrence of each stage. It was estimated that the life spans of the main germ cells were as follows: type B spermatogonia, 5.4 days; primary spermatocytes, 16.7 days; secondary spermatocytes, 0.7 days; and spermatids, 21.4 days. The results suggest that the kinetics of spermatogenesis in marsupials show a similar pattern to that in eutherians.

scholarly journals Preovulatory follicle development and ovulation in the brushtail possum (Trichosurus vulpecula) monitored by repeated laparoscopy

Reproduction ◽  
1997 ◽  
Vol 110 (2) ◽  
pp. 361-370 ◽  
Author(s):  
J. L. Crawford ◽  
G. H. Shackell ◽  
E. G. Thompson ◽  
B. J. McLeod ◽  
P. R. Hurst
Keyword(s):  
Brushtail Possum ◽  
Follicle Development ◽  
Trichosurus Vulpecula ◽  
Preovulatory Follicle
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