Telomerase Activity in Somatic Tissues and Ovaries of Diploid and Triploid Rainbow Trout (Oncorhynchus mykiss) Females.

Telomerase activity has been found in the somatic tissues of rainbow trout. The enzyme is essential for maintaining telomere length but also assures homeostasis of the fish organs, playing an important role during tissue regeneration. The unique morphological and physiological characteristics of triploid rainbow trout, when compared to diploid specimens, make them a promising model for studies concerning telomerase activity. Thus, in this study, we examined the expression of the Tert gene in various organs of subadult and adult diploid and triploid rainbow trout females. Upregulated Tert mRNA transcription was observed in all the examined somatic tissues sampled from the triploid fish when compared to diploid individuals. Contrastingly, Tert expression in the ovaries was significantly decreased in the triploid specimens. Within the diploids, the highest expression of Tert was observed in the liver and in the ovaries of the subadult individuals. In the triploids, Tert expression was increased in the somatic tissues, while the ovaries exhibited lower activity of telomerase compared to other organs and decreased compared to the ovaries in the diploids. The ovaries of triploid individuals were underdeveloped, consisting of only a few oocytes. The lack of germ cells, which are usually characterized by high Tert expression, might be responsible for the decrease in telomerase activity in the triploid ovaries. The increase in Tert expression in triploid somatic tissues suggests that they require higher telomerase activity to cope with environmental stress and maintain internal homeostasis.

Telomerase Activity in Androgenetic Rainbow Trout with Growth Deficiency and in Normally Developed Individuals.

Role of telomerase in specimens with retarded growth (dwarfs) has not been thoroughly examined to date. Considering that some of the fish species show correlation between somatic growth and activity of telomerase, it has been tempting to assume that pattern of telomerase activity in specimens with retarded growth and these with normal growth rate may vary. In the present research, telomerase activity has been examined in liver, skin, and muscles in the androgenetic rainbow trout (Oncorhynchus mykiss) with growth deficiency and their normally developed siblings. Among the examined organs, the liver showed the highest telomerase activity in all studied fish, what may be linked to the enormous regeneration capacity of the liver tissue. Although dwarf specimens examined here displayed significantly lower body size and weight they did not exhibit any significant differences in the telomerase activity measured in liver and muscle when compared to the rainbow trout without growth deficiency. In turn, telomerase activity in skin was significantly upregulated in the normally developed androgenotes. The present study indicates that dwarfism in the androgenetic rainbow trout is neither associated with ceased telomerase activity nor its decrease throughout the ontogenetic development.

Application of high hydrostatic pressure (HHP) shock to induce triploid development in the European grayling (Thymallus thymallus L.).

Use of parental individuals from allochthonous lineages to produce stocking specimens resulted in genetic contamination of many local European grayling (Thymallus thymallus) populations. Stocking programs including triploid individuals that are not able to reproduce with stocks located in natural habitats due to being reproductively sterile could contribute to populations of the European grayling and prevent regional lineages from the genetic pollution. The main goal of the present research, therefore, was to provide conditions for triploidization of the European grayling and examine gonads of triploid individuals. High Hydrostatic Pressure (HHP) shock (9000 psi/5 min.) applied to activated and incubated grayling eggs at 10 °C between the 17 min. 30 sec. and 20 minute timepoints subsequent to the time of insemination resulted in inhibition of the second polar body release and triploidization. Histological analysis confirmed that all 1-yr-old cytogenetically confirmed triploids had gonadal tissues indicative of sterility with the gonads being composed of connective tissue including fibroblasts, adipocytes and degenerated epithelial structures without there being any differentiated germ cells detected. There were no differences in the body length and Fulton's condition factor between 1 year old diploid and triploid grayling, however, the average body weight was markedly less in triploid than diploid specimens. Although, 1-yr-old triploid European female and male graylings did not have fully developed gonads, before these triploid specimens are deemed safe, as a result of being reproductively sterile for stocking in natural grayling habitats, there needs to be further examination of the 2- and 3-year-old triploid specimens.

Telomere length variation does not correspond with the growth disturbances in the rainbow trout (Oncorhynchus mykiss).

Somatic growth is considered to affect pace of the telomere attrition in vertebrates. As normally developed and dwarf fish differ in the body size we have decided to compare telomere length in the rainbow trout (Oncorhynchus mykiss) with normal growth and with growth reduced due to the dwarf condition. Examined 1-year-old fish with normal and dwarf appearance were siblings originated from androgenetic fully homozygous doubled haploid (DH) line of rainbow trout. Particular dwarf individuals had body deformities such as humpback, kyphosis, and lordosis. Somatic cells of examined rainbow trout had an average telomere length between 17 and 20 kb, comparable in females and males. Dwarf rainbow trout exhibited significantly lower body length and weight than their normally developed siblings even though no differences in the telomere length were found between these fishes. Statistical analysis did not exhibit any correlation between body size and the telomere length. Equal length of telomeres observed in the studied normal and dwarf rainbow trout suggests morphological and physiological differences in fish with different growth rates do not affect dynamics of telomeric DNA. Or any variation in the telomere length might have been levelled by telomerase that in rainbow trout is active in all tissues irrespective of the individual developmental stage.

Telomere Dynamics in the Diploid and Triploid Rainbow Trout (Oncorhynchus mykiss) Assessed by Q-FISH Analysis.

Changes of telomere length with age were assessed in diploid and triploid rainbow trout (Oncorhynchus mykiss) females in the cross-sectional study using Q-FISH technique. Triploid trout as sterile do not invest an energy in gametogenesis and continue to grow, whereas fertile diploid individuals suffer from declines in growth and survival during sexual maturation. However, triploid and diploid specimens exhibited similar patterns of telomere dynamics. Telomere length in the embryos, larvae and one-year-old juveniles did not change significantly. In the second year after hatching, subadults exhibited substantially shortened telomeres, while significant increase of the telomere length was reported in the three-year-old adults. On the other hand, correlation between telomere length and body size was observed in the triploid, but not in the diploid rainbow trout. Telomere shortening observed in two-year-old subadults may have been associated with the premature period of the fast growth in rainbow trout. Similar pattern of the telomere dynamics reported in the fertile diploids and sterile triploids indicated processes related to reproduction did not affect telomere dynamics in this species. Unexpected increase of the telomere length reported during the third year of life confirmed that in rainbow trout telomeric DNA shortens and lengthens, depending on the developmental stage.