Karyotype variation in the albino rainbow trout (Oncorhynchus mykiss (Walbaum))

Genome ◽  
2009 ◽  
Vol 52 (4) ◽  
pp. 347-352 ◽  
Author(s):  
K. Ocalewicz ◽  
S. Dobosz

A Robertsonian polymorphism resulting in diploid chromosome number ranging from 59 to 61 and constant chromosome arm number (fundamental number = 104) was observed in the albino rainbow trout ( Oncorhynchus mykiss (Walbaum)) from the yellow color strain. In one individual, 90 mitotic chromosomes and 156 chromosome arms were counted, indicating the fish as a triploid. Morphology of the chromosomes, DAPI staining, and the cytogenetic location of 5S rDNA sequences showed sex-related chromosomal heteromorphism in the specimens. Additionally, length polymorphism of the X chromosome was detected in the studied individuals and two morphs of the X chromosome were described, XL and XS, according to the size of its short arm (p). The XS was observed in the female as well as male albino rainbow trout; however, among females, no XSXS genotype was found. After primed in situ labeling with 5S rDNA primers, the p-arms of both types of the X chromosome showed similar hybridization signals. On the other hand, fluorescence in situ hybridization with telomeric PNA (peptide nucleic acid) probe exhibited weak hybridization spots on the p-arm of the XS chromosome compared with the distinct hybridization spots observed on the XL p-arm. This could reflect a different telomere length on the p-arm of the XS and XL chromosomes. Partial translocation and deletion of the X chromosome p-arm are considered to be responsible for the p-arm length difference between the two morphological variants of X chromosome.

1997 ◽  
Vol 34 (3) ◽  
pp. 253-261 ◽  
Author(s):  
Ø. Evensen ◽  
N. J. Olesen

The influence of the primary antibody, the fixative, and the antigen unmasking technique on the method sensitivity of immunohistochemistry as a method for the identification of viral hemorrhagic septicemia (VHS) virus in paraffin-embedded specimens of naturally infected rainbow trout ( Oncorhynchus mykiss) was examined. Fish (200-300 g) were collected during an outbreak of VHS. Parallel specimens from liver, spleen, kidney, and brain were fixed by immersion in 10% phosphate-buffered formalin, periodate-lysine-paraformaldehyde (PLP), Bouin's fluid, or absolute ethanol. Virus cultivation was also performed on parallel specimens, and the virus titer (TCID50/ml) was determined. Purified nucleocapsid protein (N-protein) of the virus was incorporated in an artificial antigen substrate (polymerized bovine serum albumin), fixed as described above, and embedded in paraffin wax. Microwave unmasking was performed on formalin-, PLP-, and Bouin's fluidfixed specimens. The presence of virus peptides in situ or N-protein in the artificial antigen substrates was visualized using an immunohistochemical method based on alkaline phosphatase or peroxidase and one polyclonal and five monoclonal polypeptide-specific antibodies. VHS virus was identified in situ in specimens with high virus titers (107–8 TCID50/ml) regardless of the fixative and without the need of an unmasking procedure. A pronounced masking effect was observed for the cross-linking formalin and PLP fixatives. Regardless of the primary antibodies used, there was a significantly higher epidemiologic sensitivity (the proportion of virus positive samples that tested positive by immunohistochemistry) using ethanol and Bouin's fluid compared with formalin and PLP ( P < 0.05). At 105 TCID50/ml, the average sensitivity reached 0.5, and at ≥106 TCID50/ml, sensitivity was 0.9. Unmasking procedures showed a moderate effect and did not result in significantly higher epidemiologic sensitivity ( P = 0.17). There was great variation for the different monoclonal antibodies/antigens and fixatives. Sensitivity studies on antigen substrates were in accordance with results of in situ studies that showed the highest sensitivity for ethanol and Bouin's fluid. Virus cultivation was more sensitive than immunohistochemistry. This study showed that the fixative and the primary antibody both influence method sensitivity and that VHS virus antigens concealed during fixation are difficult to reexpose. Immunostaining for VHS virus should be performed with monoclonal antibodies specific for the N-protein, and tissue samples should be fixed in either ethanol or Bouin's fluid. Immunohistochemistry is specific but is less sensitive than virus cultivation. Immunostaining for VHS virus can be a valuable supplement to virus cultivation during acute outbreaks of disease.


2008 ◽  
Vol 295 (3) ◽  
pp. R971-R978 ◽  
Author(s):  
Goran Ivanis ◽  
Marvin Braun ◽  
Steve F. Perry

Experiments were performed to assess the possible involvement of the Na+/H+ exchanger isoform 3 (NHE3; SLC9A3) in renal acid-base regulation in adult rainbow trout ( Oncorhynchus mykiss). NHE3 mRNA was expressed at high levels in the kidney relative to its paralog, NHE2. The results of in situ hybridization demonstrated an abundance of NHE3 mRNA in renal tubules. The combination of immunocytochemistry and histological staining revealed that NHE3 was confined to the apical membrane of proximal tubules, where it was colocalized with the vacuolar-type H+-ATPase. Levels of NHE3 protein (assessed by Western blotting) were increased during hypercapnia, likely as a result of increased transcription, as indicated by increasing levels of NHE3 mRNA (as determined by real-time PCR). Plasma cortisol concentration was increased during hypercapnia, and administration of exogenous cortisol caused a marked increase in NHE3 mRNA and protein. Thus we speculate that the elevation of plasma cortisol during hypercapnia contributes to transcriptional activation of NHE3 that ultimately promotes acid-base regulation by stimulating H+ secretion and HCO3− reabsorption.


2020 ◽  
Vol 269 ◽  
pp. 114675
Author(s):  
Waly Ndianco Ndiaye ◽  
Marie-Hélène Deschamps ◽  
Yves Comeau ◽  
Kabir Chowdhury ◽  
Jean-Daniel Bunod ◽  
...  

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