BACKGROUND: Huntington disease (HD) is a fatal neurodegenerative disorder involving reduced muscle coordination, mental and behavioral changes, and testicular degeneration. In order to further clarify the decreased fertility and penetration ability of the spermatozoa of transgenic HD minipig boars (TgHD), we applied a set of mitochondrial metabolism (MM) parameter measurements to this promising biological material, which can be collected noninvasively in longitudinal studies. OBJECTIVE: We aimed to optimize methods for MM measurements in spermatozoa and to establish possible biomarkers of HD in TgHD spermatozoa expressing the N-terminal part of mutated human huntingtin. METHODS: Semen samples from 12 TgHD and wild-type animals, aged 12-65 months, were obtained repeatedly during the study. Respiration was measured by polarography, MM was assessed by the detection of oxidation of radiolabeled substrates (mitochondrial energy-generating system; MEGS), and the content of the oxidative phosphorylation system subunits was detected by Western blot. Three possibly interfering factors were statistically analyzed: the effect of HD, generation and aging. RESULTS: We found 5 MM parameters which were significantly diminished in TgHD spermatozoa and propose 3 specific MEGS incubations and complex I-dependent respiration as potential biomarkers of HD in TgHD spermatozoa. CONCLUSIONS: Our results suggest a link between the gain of toxic function of mutated huntingtin in TgHD spermatozoa and the observed MM and/or glycolytic impairment. We determined 4 biomarkers useful for HD phenotyping and experimental therapy monitoring studies in TgHD minipigs.
- MeSH
- dýchání MeSH
- geneticky modifikovaná zvířata MeSH
- Huntingtonova nemoc komplikace genetika patologie MeSH
- kyseliny trikarboxylové metabolismus MeSH
- lidé MeSH
- miniaturní prasata MeSH
- mitochondriální proteiny metabolismus MeSH
- mitochondrie metabolismus MeSH
- mutace genetika MeSH
- oxidativní fosforylace MeSH
- prasata MeSH
- protein huntingtin genetika MeSH
- pyruvátdehydrogenasový komplex metabolismus MeSH
- sperma metabolismus MeSH
- spermie metabolismus patologie MeSH
- trinukleotidové repetice genetika MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Huntington's disease is induced by CAG expansion in a single gene coding the huntingtin protein. The mutated huntingtin (mtHtt) primarily causes degeneration of neurons in the brain, but it also affects peripheral tissues, including testes. OBJECTIVE: We studied sperm and testes of transgenic boars expressing the N-terminal region of human mtHtt. METHODS: In this study, measures of reproductive parameters and electron microscopy (EM) images of spermatozoa and testes of transgenic (TgHD) and wild-type (WT) boars of F1 (24-48 months old) and F2 (12-36 months old) generations were compared. In addition, immunofluorescence, immunohistochemistry, Western blot, hormonal analysis and whole-genome sequencing were done in order to elucidate the effects of mtHtt. RESULTS: Evidence for fertility failure of both TgHD generations was observed at the age of 13 months. Reproductive parameters declined and progressively worsened with age. EM revealed numerous pathological features in sperm tails and in testicular epithelium from 24- and 36-month-old TgHD boars. Moreover, immunohistochemistry confirmed significantly lower proliferation activity of spermatogonia in transgenic testes. mtHtt was highly expressed in spermatozoa and testes of TgHD boars and localized in all cells of seminiferous tubules. Levels of fertility-related hormones did not differ in TgHD and WT siblings. Genome analysis confirmed that insertion of the lentiviral construct did not interrupt any coding sequence in the pig genome. CONCLUSIONS: The sperm and testicular degeneration of TgHD boars is caused by gain-of-function of the highly expressed mtHtt.
- MeSH
- genetické vektory MeSH
- geneticky modifikovaná zvířata MeSH
- Huntingtonova nemoc metabolismus patologie MeSH
- Lentivirus genetika MeSH
- lidé MeSH
- miniaturní prasata MeSH
- modely nemocí na zvířatech MeSH
- mutace * MeSH
- počet spermií MeSH
- prasata MeSH
- proliferace buněk fyziologie MeSH
- protein huntingtin genetika metabolismus MeSH
- spermie metabolismus patologie MeSH
- stárnutí metabolismus patologie MeSH
- testis metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- Huntingtonova nemoc * diagnóza etiologie genetika MeSH
- interakce spermie a vajíčka MeSH
- kapacitace spermií MeSH
- laboratorní zvířata MeSH
- lidé MeSH
- mitochondrie * fyziologie metabolismus patologie ultrastruktura MeSH
- mužská infertilita MeSH
- myši MeSH
- odběr biologického vzorku * využití MeSH
- oxidativní fosforylace * MeSH
- preklinické hodnocení léčiv MeSH
- spermie * abnormality fyziologie metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Pri Huntingtonovej chorobe dochádza ktestikulárnej degenerácii. Tá je následne príčinou zníženej reprodukčnej schopnosti pacientov. Do teraz ale nie je zistená presná príčina spôsobujúca poruchu spermiogenézy. Následnými štúdiami by sa mohla táto príčina objasniť a tým taktiež napomôcť k objasneniu patogenézy mutovaného huntingtínu.
One of the effect of Huntington‘s disease is the testicular degeneration. It causes a reduced reproductive capacity of patients. Unfortunately, up to now the exact cause of the malfunction of spermiogenesis is not know. Subsequent studies could clarify the cause and thus help to clarify the pathogenesis of mutated huntingtin.
- MeSH
- Huntingtonova nemoc * komplikace patologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- mužská infertilita * diagnóza genetika patologie MeSH
- mužské pohlavní orgány patologie MeSH
- prasata MeSH
- spermatogeneze MeSH
- spermie patologie MeSH
- testis patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
