Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis)

Abstract

To understand mechanisms of spermatogenesis, the proteome and the phosphoproteome in prepubertal and pubertal swamp buffalo (Bubalus bubalis) testes were analyzed using tandem mass tag (TMT) coupled with liquid chromatography-tandem mass spectrometry (LC–MS/MS). In prepubertal testes, 80 proteins were overexpressed, 148 proteins were underexpressed, and 139 and 142 protein sites had higher and lower phosphorylation, respectively, compared to the levels in pubertal testes. Several of these proteins were associated with reproductive processes such as sexual reproduction, spermatogenesis, fertilization, and spermatid development. In particular, outer dense fiber protein 1 (ODF1), protein maelstrom homolog (MAEL), actin-like protein 7B (ACTL7B), tyrosine-(Y)-phosphorylation regulated (CABYR), and tripartite motif containing 36 (TRIM36) were upregulated with age at both the proteome and phosphoproteome levels. Combining proteome and phosphoproteome analysis can be effectively applied to study the protein/phosphorylation patterns of buffalo testes. These data provide new regulatory candidates and evidence for a complex network in spermatogenesis in buffalo testes, and serve as an important resource for exploring the physiological mechanism of spermatogenesis in mammals.

Microtubular TRIM36 E3 Ubiquitin Ligase in Embryonic Development and Spermatogenesis

“…In addition, they showed that retinoic acid, directly involved in spermatogenesis developmental transitions [ 34 , 40 ], can anticipate the expression of TRIM36 early on culture (day 8) [ 41 ]. Further, differential proteomics and phosphoproteomics in buffalo, comparing pre-pubertal and pubertal testes, demonstrated that TRIM36 is upregulated with age suggesting a role in the spermatogenesis process in this species [ 42 ]. Furthermore, Trim36 null mouse-derived spermatozoa showed lower total and progressive motility and less complex motility patterns [ 27 ].…”

Actl7b-deficiency leads to mislocalization of LC8 type dynein light chains and disruption of murine spermatogenesis

“…Further, proteomic and phosphoproteomic analysis of prepubertal and pubertal testis of swamp buffalo identified ACTL7B to be higher abundant and phosphorylated in the pubertal testis, again suggesting a role in spermatogenesis (Huang et al, 2020). Single nucleotide polymorphisms in the coding sequence of ACTL7B in infertile men have been reported, however not been directly correlated to male infertility (Tanaka et al, 2007, Tanaka et al, 2019.…”

Actl7b deficiency leads to mislocalization of LC8 type dynein light chains and disruption of murine spermatogenesis

“…Noteworthy, a study comparing two groups of Luzhong mutton sheep with different fecundity identified nine genes, among these ACTL7B (and ACTL7A), which were associated with reduced litter size (Tao et al, 2021). Further, proteomic and phosphoproteomic analysis of prepubertal and pubertal testis of swamp buffalo identified ACTL7B to be higher abundant and phosphorylated in the pubertal testis, again suggesting a role in spermatogenesis (Huang et al, 2020). Single nucleotide polymorphisms in the coding sequence of ACTL7B in Development • Accepted manuscript infertile men have been reported, however not been directly correlated to male infertility (Tanaka et al, 2007, Tanaka et al, 2019.…”