Large-scale discovery of male reproductive tract-specific genes through analysis of RNA-seq datasets

Abstract

Background: The development of a safe, effective, reversible, non-hormonal contraceptive method for men has been an ongoing effort for the past few decades. However, despite significant progress on elucidating the function of key proteins involved in reproduction, understanding male reproductive physiology is limited by incomplete information on the genes expressed in reproductive tissues, and no contraceptive targets have so far reached clinical trials. To advance product development, further identification of novel reproductive tract-specific genes leading to potentially druggable protein targets is imperative. Results: In this study, we expand on previous single tissue, single species studies by integrating analysis of publicly available human and mouse RNA-seq datasets whose initial published purpose was not focused on identifying male reproductive tract-specific targets. We also incorporate analysis of additional newly acquired human and mouse testis and epididymis samples to increase the number of targets identified. We detected a combined total of 1178 genes for which no previous evidence of male reproductive tract-specific expression was annotated, many of which are potentially druggable targets. Through RT-PCR, we confirmed the reproductive tract-specific expression of 51 novel orthologous human and mouse genes without a reported mouse model. Of these, we ablated four epididymis-specific genes (Spint3, Spint4, Spint5, and Ces5a) and two testis-specific genes (Pp2d1 and Saxo1) in individual or double knockout mice generated through the CRISPR/Cas9 system. Our results validate a functional requirement for Spint4/5 and Ces5a in male mouse fertility, while demonstrating that Spint3, Pp2d1, and Saxo1 are each individually dispensable for male mouse fertility. Conclusions: Our work provides a plethora of novel testis-and epididymis-specific genes and elucidates the functional requirement of several of these genes, which is essential towards understanding the etiology of male infertility and the development of male contraceptives.

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

“…Our study generated a high-quality transcriptomic data for testicular tissue of adult and juvenile domestic cats, which provides an important resource for future studies on testis development, spermatogenesis and fertility in cats, as well as new insights into tissue resilience. Transcriptomic data can also contribute to identification of druggable protein targets in male reproductive tracts and development of male contraception [ 59 ]. Our study provides a 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.…”

Transcriptomic analysis of the seminal vesicle response to the reproductive toxicant acrylamide

“…To compare the specificity of the seminal vesicle transcriptome with that of other mouse tissues, we interrogated data from a recent large scale discovery study of male reproductive tract specific genes [33] (Fig. 2B).…”

“…To compare the seminal vesicle transcriptome to previously published mouse transcriptomic analyses, we obtained data from a recent large-scale discovery study of male reproductive tract specific genes [33], which presented data in TPM. These transcriptomic data included testis, spermatogonia, Sertoli cell, caput epididymis, corpus epididymis, cauda epididymis, as well as the transcriptomes of non-reproductive tissues, including adipose tissue, adrenal gland, bone marrow, cerebral cortex, colon, duodenum, heart, kidney, liver, lung, pancreas, skeletal muscle, small intestine and stomach [33,[124][125][126][127] and were log2 TPM data collated from Additional file 4, Table S4 in [33]. From our seminal vesicle transcriptome, RSEM calculated autosomal gene expression levels [120] were normalised using the TPM approach [120] and for the comparison to other cell types was similarly converted to log2 values.…”

“…gov/geo/query/acc.cgi?acc=GSE183632. Other transcriptomics data used in this study were obtained from [16] with permission from the authors, and accessed from Additional file 4, Table S4 of [33].…”

Análise da expressão de genes alvos em homens com varicocele

“…Os genes WFDC são importantes durante a maturação espermática e na formação de proteínas que participam da coagulação seminal, além de serem altamente expressos no epidídimo (CLAUSS et al, 2011). Não o bastante, foi visto que os genes SPINT3 e o SPINT4 são expressos no epidídimo com alta especificidade também, o que sugere um papel na maturação dos espermatozoides (MCCRUDDEN et al, 2008;ROBERTSON et al, 2020). O aumento da expressão de SPINT3 pode ser reflexo do aumento da temperatura escrotal, conduzindo assim, a um desiquilíbrio das proteínas presentes no sêmen.…”