‘Spontaneous’ sex reversal in organ cultures of the embryonic male gonad of the bird

Development ◽  
1974 ◽  
Vol 31 (3) ◽  
pp. 611-620
Author(s):  
Gregory F. Erickson
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Calf Serum ◽  
Sex Reversal ◽  
Biologically Active ◽  
Foetal Calf Serum ◽  
Developmental Sequence ◽  
Organ Cultures ◽  
Male Gonad ◽  
Ovarian Cortex

The left embryonic testis of the bird (4–8 days of incubation) was organ cultured in medium that contained 10% foetal calf serum. Under these conditions, the germinal epithelium (GE) of the 4-day gonad differentiates into an ovarian cortex and the male primordial germ cells (PGCs) complete a developmental sequence similar to normal oocytes, i.e. they divide mitotically, develop a Balbiani body, divide synchronously in groups of two, four, and eight germ cells, and some enter pre-leptotene. No medullary tissue develops in the 4-day explants. The pieces of 6- and 8-day gonad differentiate into true ovotestes in which the GE develops into a cortex and the medulla develops into seminiferous cords. The PGCs in the cortex differentiate as oocytes and those in the seminiferous cords differentiate as spermatogonia. The possibility that biologically active oestrogens are present in the growth medium is discussed.

Response of preimplantation rat blastocysts in vitro to extracellular uterine luminal components, serum and hormones

1977 ◽  
Vol 25 (1) ◽  
pp. 265-277
Author(s):  
M.A. Surani
Keyword(s):  
Calf Serum ◽  
Giant Cells ◽  
Biologically Active ◽  
Foetal Calf Serum ◽  
Active Components ◽  
Rat Serum ◽  
Pregnant Females ◽  
Trophoblast Giant Cells

The influence of extracellular environmental factors on preimplantation rat blastocysts was tested by determining the number of embryos which escaped from their zonae pellucidae, followed by attachment and outgrowth of trophoblast giant cells, after 72 h in culture Uterine luminal ocmponents from individual females, or hormones, were included in Dulbecco's medium which contained 4 mg/ml bovine serum albumin. In about 20% of cases, uterine fluids were embryotonic. However, uterine fluids from day-5 pregnant females, the day of implantation in the rat, were more potent in these tests than uterine fluids obtained from ovariectomized females treated with progesterone alone. The potency of a mixture of the 2 fluids was also high. Uterine fluids obtained at 14 h after an injection of oestradiol and progesterone to the ovariectomized females, were also effective in these tests. Rat serum and foetal calf serum were effective too, but steroids or insulin alone in the medium had no detectable influence on embryos. Serum or uterine luminal proteins appear to be essential for maintaining the viability of the blastocysts and for inducing the responses observed here. In the uterine fluids, some proteins released into the lumen after treatment of females with oestradiol and progesterone appear to be the biologically active components. Differences in the responses of blastocysts in vitro are compared with those in vivo.

scholarly journals Complete developmental cycle ofLeishmania mexicanain axenic culture

Parasitology ◽  
1994 ◽  
Vol 108 (1) ◽  
pp. 1-9 ◽  
Author(s):  
P. A. Bates
Keyword(s):  
Calf Serum ◽  
Axenic Culture ◽  
Biochemical Properties ◽  
Total Protein Content ◽  
Developmental Cycle ◽  
Foetal Calf Serum ◽  
Developmental Sequence ◽  
Metacyclic Promastigotes ◽  
Secretory Acid Phosphatase ◽  
First Time

SUMMARYA complete developmental sequence ofLeishmania mexicanahas been produced in axenic culture for the first time. This was achieved by manipulation of media, pH and temperature conditions over a period of 16 days. All experiments were initiated with lesion amastigotes that were transformed to multiplicative promastigotes by culture in HOMEM, 10% foetal calf serum, pH 7·5, at 25 °C. Metacyclogenesis was induced by subpassage in Schneider'sDrosophila medium, 20% foetal calf serum, pH 5·5, and the resulting forms transformed to axenically growing amastigotes by subpassage in the same medium and raising the temperature to 32 °C. Parasites from each day were characterized with respect to their general morphology using light microscopy of Giemsa-stained smears, and biochemically by analysis of total protein content, proteinases, nucleases and secretory acid phosphatase. The results demonstrated that the three main stages identified - amastigotes, multiplicative promastigotes and metacyclic promastigotes - each exhibited the expected suite of biochemical properties. Further, the changes in morphology observed as the developmental sequence proceeded from stage to stage were accompanied by appropriate changes in biochemical properties. These results provide both useful biochemical markers and a culture system in which to examine the regulation of differentiation and transformation during theLeishmanialife-cycle.

scholarly journals Association of culture of mouse urogenital complexes in media containing rodent sera with the appearance of primordial germ cell-like cells

Reproduction ◽  
2003 ◽  
pp. 519-526 ◽  
Author(s):  
T Mayanagi ◽  
K Ito ◽  
J Takahashi
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Calf Serum ◽  
Culture Method ◽  
Embryonic Germ Cells ◽  
Alkaline Phosphatase Staining

Primordial germ cells differentiate into germ cells and have the ability to reacquire totipotency. Mouse primordial germ cells are identified by alkaline phosphatase staining of the extraembryonic mesoderm, and they proliferate and migrate to reach the genital ridges. Mouse primordial germ cells have never been maintained in culture exclusively for longer than a week without differentiation or dedifferentiation. Moreover, primordial germ cells have not been proliferated with urogenital complexes in vitro, because gonad culture has never been successful. It was thought that primordial germ cells could proliferate in a culture of urogenital complex under modified medium conditions resembling those in vivo; however, organ culture of mouse gonad has been performed with fetal calf serum or equine serum, and those sera produce conditions different from those in vivo. Therefore, mouse urogenital complexes were cultured in media containing rodent sera. As a result, it was possible to proliferate primordial germ cell-like cells outside gonads, and these cells very closely resembled primordial germ cells. In addition, motile primordial germ cell-like cells could be obtained. The ability to maintain primordial germ cell-like cells in culture by this intra-species culture method is important in the study of gametogenesis. Furthermore, this method is useful as a source of stem cells such as embryonic germ cells.

Germ-cell transfer and sex ratio in Xenopus laevis

Development ◽  
1965 ◽  
Vol 13 (1) ◽  
pp. 51-61
Author(s):  
A. W. Blackler
Keyword(s):  
Xenopus Laevis ◽  
Germ Cell ◽  
South African ◽  
Germ Cells ◽  
Sexual Differentiation ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Hormone Treatment ◽  
Sex Reversal ◽  
Cell Transfer

A Technique for the transfer of primordial germ cells between neurulae of the South African Clawed Toad Xenopus laevis has been described by Blackler & Fischberg (1961). This method was originally developed with the object in mind of eventually making a genetic analysis of abnormal embryos resulting from the transplantation of somatic nuclei. Such analysis involves two schemes which require the transfer of embryonic gonocytes from the defective transplant embryo to a normal recipient. Moreover, one of these two schemes requires that transferred germ cells be reversed in their sexual differentiation in the developing gonad of the host (see Fischberg, 1961; Fischberg & Blackler, 1963a, b). Since it has been known for some time, from experiments involving parabiosis, transplantation of the gonadal rudiment and hormone treatment (e.g. Burns, 1925, 1930; Witschi, 1937; Humphrey, 1929, 1933, 1948, 1957; Gallien, 1953, 1956), that the manifestation of the sex genotype of a primordial germ cell can be physiologically reversed by the hormonal characteristics of the gonad, there seemed no obstacle to obtaining sex-reversal of the transferred gonocytes in Xenopus.

Development in vitro of the female germ cells of the polychaete Ophryotrocha labronica

Development ◽  
1985 ◽  
Vol 85 (1) ◽  
pp. 151-161
Author(s):  
Hadar Emanuelsson ◽  
Siw Anehus
Keyword(s):  
Amino Acids ◽  
Germ Cells ◽  
Nurse Cell ◽  
Sea Water ◽  
Calf Serum ◽  
Growth Period ◽  
Foetal Calf Serum ◽  
Leading Role ◽  
Development In Vitro

In the polychaete Ophryotrocha labronica each oocyte is during its growth period associated with a single nurse cell. The fact that the oocyte-nurse cell pairs occur isolated in the female coelom makes them easily removable for analysis of their developmental ability in vitro. Using Dulbecco's modified Eagle medium supplemented with amino acids, nucleosides, foetal calf serum and sea water, we have managed to support development in vitro of germ cell pairs from early and mid-oogenesis until maturation of the oocyte, when the nurse cell degenerates and the oocyte enters meiotic metaphase. Radiolabelling of germ cells in mid-oogenesis with tritiated amino acids and uridine during the first day of incubation indicates normal development with synthesis of RNA and protein, and pulses two days later verify a continued normal protein synthesis and yolk formation. The investigation confirms autosynthesis of yolk proteins in the germ cells of this species and indicates a leading role of the nurse cell in the process.

THE EFFECT OF PROGESTERONE AND OESTRADIOL ON IMMATURE MOUSE UTERI MAINTAINED AS ORGAN CULTURES

1973 ◽  
Vol 57 (1) ◽  
pp. 171-174 ◽  
Author(s):  
P. S. GRANT
Keyword(s):  
Calf Serum ◽  
Foetal Calf Serum ◽  
Organ Cultures ◽  
Independent Observer ◽  
Histological Sections ◽  
Progesterone Treatment ◽  
Immature Mouse

SUMMARY Uterine horns from immature mice were cultured for 2 days in Trowell's T8 medium containing 20% foetal calf serum and either ethanol (0·125%), oestradiol-17β (5 μg/ml), progesterone (5 μg/ml) or progesterone plus oestradiol (5 μg + 5 μg/ml). The uteri were fixed in Bouin's fluid and the degree of closure of the lumen was assessed by an independent observer after examination of histological sections. Uteri treated with oestradiol alone usually did not differ from those treated with ethanol: progesterone treatment either alone or in combination with oestradiol led to luminal closure.

Hexachlorobenzene toxicity in the rat ovary: II. Ultrastructure induced by medium (10 mg/kg) dose exposure

1992 ◽  
Vol 50 (1) ◽  
pp. 668-669
Author(s):  
Amreek Singh ◽  
Warren G. Foster ◽  
Anna Dykeman ◽  
David C. Villeneuve
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Surface Epithelium ◽  
Morphological Parameters ◽  
Comprehensive Investigation ◽  
Ovary Surface Epithelium ◽  
Rat Ovary ◽  
High Doses

Hexachlorobenzene (HCB) is a known toxicant that is found in the environment as a by-product during manufacture of certain pesticides. This chlorinated chemical has been isolated from many tissues including ovary. When administered in high doses, HCB causes degeneration of primordial germ cells and ovary surface epithelium in sub-human primates. A purpose of this experiment was to determine a no-effect dose of the chemical on the rat ovary. The study is part of a comprehensive investigation on the effects of the compound on the biochemical, hematological, and morphological parameters in the monkey and rat.

Transfusion of Chick Primordial Germ Cells into Quail Embryos and their Settlement in Gonads

1998 ◽  
Vol 69 (10) ◽  
pp. 911-915 ◽  
Author(s):  
Tamao ONO ◽  
Ryohei YOKOI ◽  
Seishi MAEDA ◽  
Takao NISHIDA ◽  
Hirohiko AOYAMA
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells
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