Testis differentiation in anuran amphibians is the result of two opposing processes: degeneration of the distal part, and development of the proximal part, which becomes a functional male gonad. Undifferentiated gonad differentiates directly into a testis without a transition phase. We described the morphology of developing testes in Rana temporaria and Hyla arborea, and made careful histology and ultrastructure in Pelophylax lessonae. The developing testis was divided into 10 stages (I-III, undifferentiated gonad, IV-X, testis). The earliest morphological symptoms of testis differentiation were observed in 4- to 5-week-old tadpoles at Gosner stage 27-28. At that time an undifferentiated gonad, composed of 6-9 metameres, differentiates into a testis. The proximal metameres (2-3 in the right gonad and 3-4 in the left one) differentiate into a functional testis, while the distal ones degenerate. The difference between left and right gonad size is maintained until adulthood (stage X). Degeneration of the distal part progresses along the posterior-anterior gradient and starts at stage IV. It affects first the germ cells with accompanying precursor Sertoli cells, and then the mesenchymal cells and fibroblasts. Finally the external epithelium forms a "sleeve" around the almost empty distal part. The total length of the testes stays the same until stage VIII at Gosner stage 41 (age 74-148 days). Active spermatogenesis starts at stage IX (juveniles after their first hibernation), during which the distal part eventually disappears and the proximal part starts growing considerably due to progressing spermatogenesis.
DNA elimination is a radical form of gene silencing and occurs both in somatic and germ cells. The programmed DNA elimination occurs during gametogenesis in interspecies hybrids that reproduce by hybridogenesis (stick insects, fishes, and amphibians) and concerns removal of whole genomes of one of the parental species and production of clonal gametes propagating the genome of the other species. The cellular mechanisms differ considerably in hybridogenetic insects and fishes but remains unknown in edible frogs Pelophylax esculentus, natural hybrids between Pelophylax lessonae and Pelophylax ridibundus. Here we report DNA elimination mechanism in early developing gonads of diploid and triploid hybrid frogs, studied by TEM, immunofluorescence, and cytochemistry. In gonocytes of both sexes (primary oogonia and prespermatogonia), micronuclei emerge as detached nuclear buds formed during interphase. We found depletion of nuclear pore complexes in micronuclear membrane and chromatin inactivation via heterochromatinization followed by degradation of micronuclei by autophagy. Micronuclei formation does not lead to apoptotic cell death showing that genome elimination is a physiological process. Chromatin elimination via micronuclei in P. esculentus is unique among hybridogenetic animals and contributes to broadening the knowledge about reproductive modes in animals.
The protective effects of carvedilol on heart function observed in the acute phase of experimental autoimmune myocarditis seem to be associated with its ability to decrease MMP-2 activity and subsequently prevent degradation of myofilaments and release of troponin I while not related to suppression of inflammation.
Our findings indicate that the use of both antlerogenic stem cell implants and the preparations derived from the cells offer alternative approaches to those based on autologous stem cells in the biological stimulation of osteogenesis and in bone regeneration.
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