FIGURE SUMMARY
Title

Type I Diabetes in Zebrafish Reduces Sperm Quality and Increases Insulin and Glucose Transporter Transcripts

Authors
Diogo, P., Martins, G., Simão, M., Marreiros, A., Eufrásio, A.C., Cabrita, E., Gavaia, P.J.
Source
Full text @ Int. J. Mol. Sci.

Confirmation of pancreatic β-cell ablation and onset of diabetes in zebrafish males. (A) Graphic representation of the construct inserted in the Tg(ins:nfsb-mCherry) genome. (B) Observation of pancreas with lack of fluorescence in Tg(ins:nfsb-mCherry) diabetic males, with evident pancreatic β-cell ablation and onset of diabetes. (C) Observation of pancreatic fluorescence in Tg(ins:nfsb-mCherry) control males after dissection, with the presence of live β cells (white area showing magnification of the pancreas).

Gene expression of insulin metabolism markers in zebrafish sperm pools: (A) expression of insa in spermatozoa pools from AB males in control (n = 4) and metronidazole (Met) treatment (n = 4) groups and Tg(ins:nfsb-mCherry) in control (n = 5) and Met treatment (n = 7) groups; (B) expression of insra in spermatozoa pools from AB males in control (n = 4) and Met treatment (n = 4) groups and Tg(ins:nfsb-mCherry) in control (n = 7) and Met treatment (n = 7) groups; (C) expression of slc2a2 in spermatozoa pools from AB males in control (n = 4) and Met treatment (n = 4) groups and Tg(ins:nfsb-mCherry) in control (n = 5) and Met treatment (n = 6) groups. The plotted values represent means ± SD. Different letters represent statistical differences (one-way ANOVA, post hoc SNK, p < 0.05).

Spermatozoa plasma membrane viability of fresh and cryopreserved sperm from untreated Tg(ins:nfsb-mCherry) males (n = 5) and metronidazole (Met)-treated males (n = 6). The plotted values represent means ± SD. Different superscript letters represent statistical differences (independent samples t-test, p < 0.05).

Sperm motility parameters of fresh sperm obtained from untreated Tg(ins:nfsb-mCherry) males (n = 5) and metronidazole (Met)-treated males (n = 8) and of cryopreserved sperm from untreated (n = 5) and Met-treated males (n = 10). Sperm was activated, and motility parameters were recorded every 10 s for 1 min in terms of: (A) total motility (%); (B) progressive motility (%); (C) curvilinear velocity (μm/s); (D) straight-line velocity (μm/s), and (E) linearity (%). The plotted values represent means. The dashed lines represent fresh sperm and the continuous lines represent cryopreserved sperm. Sperm from untreated males is represented with a white circle and sperm from Met-treated males is represented with a dark circle.

DNA integrity in zebrafish sperm: (A) comet with high DNA fragmentation; (B) comet with low DNA fragmentation; (C) DNA fragmentation of fresh sperm obtained from untreated Tg(ins:nfsb-mCherry) males (n = 4) and metronidazole (Met)-treated males (n = 8) and of cryopreserved sperm from untreated males (n = 5) and Met-treated males (n = 6). The plotted values represent means ± SD. Different letters represent statistical differences between treatments and the asterisk represents a statistical difference between fresh and cryopreserved sperm (independent samples t-test, p < 0.05).

Hypothetic model of putative effects of diabetes on sperm quality in type I diabetes zebrafish strain [Tg(ins:nfsb-mCherry)].

Acknowledgments
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