Transcriptome dynamics in developing testes of domestic cats and impact of age on tissue resilience to cryopreservation

Abstract

Background Fundamental knowledge of cellular and molecular mechanisms in developing testicular tissues is critical to better understand gonadal biology and responses to non-physiological conditions. The objective of our study was to (1) analyze transcriptome dynamics in developing testis of the domestic cat and (2) characterize age effects on the initial response of the tissue to vitrification. Tissues from adult and juvenile cats were processed for histology, DNA integrity, and RNA sequencing analyses before and after vitrification. Results Transcriptomic findings enabled to further characterize juvenile period, distinguishing between early and late juvenile tissues. Changes in gene expression and functional pathways were extensive from early to late juvenile to adult development stages. Additionally, tissues from juvenile animals were more resilient to vitrification compared to adult counterparts, with early juvenile sample responding the least to vitrification and late juvenile sample response being closest to adult tissues. Conclusions This is the first study reporting comprehensive datasets on transcriptomic dynamic coupled with structural analysis of the cat testis according to the age and exposure to cryopreservation. It provides a comprehensive network of functional terms and pathways that are affected by age in the domestic cat and are either enriched in adult or juvenile testicular tissues.

Feline microRNAome in ovary and testis: Exploration of in-silico miRNA-mRNA networks involved in gonadal function and cellular stress response

“…One of the biggest miRNA clusters upregulated from juvenile to adult testis included 16 mature miRNAs: miR-146a-5p, -146b-5p, -15b-3p/5p, -29b-2-3p, -29c-3p, -449-3p/5p, -34c-3p/5p, -34b-3p, -191-3p/5p, -92b-3p/5p, and -200c-3p. This cluster was predicted to inhibit functions in cell-matrix adhesion, morphogenesis, Wnt signaling, immune response, stem cell pluripotency and cell migration in adult compared to early juvenile testis, all of which were reported to also be inhibited in our previous study (Amelkina et al, 2021). From this cluster, miR-34c-3p and miR-200c-3p exhibited continuous upregulation from early to late juvenile to adult period.…”

“…In the current study, we integrate previously obtained mRNA data on domestic cat ( Felis catus ) from Study 1, BioProject PRJNA662384, ( Amelkina and Comizzoli, 2020 ), and Study 2, BioProject PRJNA741252, ( Amelkina et al, 2021 ), for miRNA-mRNA analysis ( Figure 1 ). All conditions and protocols used for tissue processing and experiments were the same as in Study 1 ( Amelkina and Comizzoli, 2020 ) and 2 ( Amelkina et al, 2021 ).…”

“…It is also an important biomedical model, carrying a similarity with human genome with high degree of conserved synteny and serving as a model for ∼250 genetic diseases that are analogous to human disorders ( O’Brien et al, 2002 ; Pontius et al, 2007 ; Tamazian et al, 2014 ). Recently, our group has been focusing on elucidating transcriptome dynamics in the ovarian and testicular tissue of domestic cat to better understand molecular mechanisms behind gonadal development and response of tissues to stress caused by preservation protocols ( Amelkina and Comizzoli, 2020 ; Amelkina et al, 2021 ). We have reported major changes in mRNA expression in the ovarian tissue in response to vitrification and microwave-assisted preservation ( Amelkina and Comizzoli, 2020 ), as well as gradual shifts in transcriptome dynamics throughout testicular development ( Amelkina et al, 2021 ).…”

“…Recently, our group has been focusing on elucidating transcriptome dynamics in the ovarian and testicular tissue of domestic cat to better understand molecular mechanisms behind gonadal development and response of tissues to stress caused by preservation protocols ( Amelkina and Comizzoli, 2020 ; Amelkina et al, 2021 ). We have reported major changes in mRNA expression in the ovarian tissue in response to vitrification and microwave-assisted preservation ( Amelkina and Comizzoli, 2020 ), as well as gradual shifts in transcriptome dynamics throughout testicular development ( Amelkina et al, 2021 ). To better understand the processes observed in these gonadal tissues, we need to go further and unravel the mechanisms of gene expression regulation.…”

Key aspects of domestic cat spermatogenesis

“…Recently, characterization of changes in gene expression and functional pathways throughout domestic cat testis maturation was carried out. These molecular changes proved to be extensive throughout development permitting differentiation between early and late juvenile tissues (Amelkina et al., 2021).…”