Localization of Receptors for Sex Steroids and Pituitary Hormones in the Female Genital Duct throughout the Reproductive Cycle of a Viviparous Gymnophiona Amphibian, Typhlonectes compressicauda.

Reproduction in vertebrates is controlled by the hypothalamo-pituitary-gonadal axis, and both the sex steroid and pituitary hormones play a pivotal role in the regulation of the physiology of the oviduct and events occurring within the oviduct. Their hormonal actions are mediated through interaction with specific receptors. Our aim was to locate α and ß estrogen receptors, progesterone receptors, gonadotropin and prolactin receptors in the tissues of the oviduct of Typhlonectes compressicauda (Amphibia, Gymnophiona), in order to study the correlation between the morphological changes of the genital tract and the ovarian cycle. Immunohistochemical methods were used. We observed that sex steroids and pituitary hormones were involved in the morpho-functional regulation of oviduct, and that their cellular detection was dependent on the period of the reproductive cycle.

Immunohistochemical Localization of Calpains in the Amphibian Xenopus laevis.

Though histochemical techniques have been used for decades, they are still very important in basic research. They make it possible to work on fixed tissues and provide a large amount of information in a relatively short time and at a low cost. Here we describe methods for indirect immunohistochemistry and immunofluorescence on sections of tadpoles and tissues of adult amphibians belonging to the species Xenopus laevis. The objective is to localize calpains within tissues in order to understand their involvement in cellular processes.

Patterns of Apoptosis and Proliferation throughout the Biennial Reproductive Cycle of Viviparous Female Typhlonectes compressicauda (Amphibia, Gymnophiona).

Typhlonectes compressicauda is an aquatic gymnophionan amphibian living in South America. Its breeding cycle is linked to seasons, characterized by a regular alternation of rainy and dry seasons. During a complex biennial cycle, the female genital tract undergoes a series of alternations of increasing and decreasing, governed by equilibrium of proliferation and apoptotic phenomena. Immunohistochemical methods were used to visualize cell proliferation with the detection of Ki67 antibody, a protein present in proliferative cells; terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and Apostain were performed to detect apoptotic cells on sections of ovaries and oviducts. In ovaries, both phenomena affect the germinal nests and follicles according to the cycle period. In the oviduct, the balance was in favor of proliferation during preparation for reproduction, and in favor of apoptosis when genital ducts regress. Apoptosis and proliferation are narrowly implicated in the remodeling of the genital tract and they are accompanied by the differentiation of tissues according to the phase of the breeding cycle. These variations permit the capture of oocytes at ovulation, always at the same period, and the parturition after 6-7 months of gestation, at a period in which the newborns live with their mother, protected in burrows in the mud. During the intervening year of sexual inactivity, the female reconstitutes body reserves.

Relationships between calpains and glutamate- or kainate-induced apoptosis in Xenopus laevis tadpoles.

Cell glutamate-damage induced by overstimulation of ionotropic receptors is initiated by modification of the intracellular Ca2+ homeostasis and the concomitant activation of Ca2+-dependent cysteine proteases, the calpain and caspase families. The resultant cleavage of target molecules mediates a critical function in the execution of the cell death. In this work, we investigated relationships between the activity of calpain and glutamate-orkainate-induced apoptosis in several organs of Xenopus laevis tadpole. Animals (stage 48) were incubated for 3 hours with glutamate (30-120 mM) or kainate (0.015-0.75 mM) and the rise of both apoptosis and calpain was observed in several organs. Our results indicated that glutamate (120 mM) or kainate (0.15 mM) exposure induced cell death with apoptotic features. The toxic effects of drugs into the organs were variable. Apoptosis was probably not the only form of cell death and option of necrosis or apoptosis was depending on the stimulation degree of the receptor, i.e. the receptor type, intensity and time course of molecule exposure. The increase of ubiquitous calpain was not correlated with the peak of apoptosis, suggesting the role of calpain in cell death was complex: calpain and caspase pathways were tightly interrelated in the glutamate- or kainate-induced cell death and the contribution of calpain to another type of death than apoptosis was perhaps preferred.