Characterization of GALNTL5 gene sequence and expression in ovine testes and sperm.

The polypeptide N-acetylgalactosaminyltransferase-like protein 5 (GALNTL5), which belongs to the polypeptide N-acetylgalactosaminyltransferase (pp-GalNAc-T) gene family, is a newly identified gene that is specifically expressed in testis and involved in spermatogenesis. However, there is no data showing the existence of GALNTL5 in ram testis at various developmental stages and its influence on sperm motility. Therefore, the objectives of the present study were to evaluate the presence of GALNTL5 in the testis of 3-24 months (M) ram and to investigate the expression of GALNTL5 in spermatozoa with different motilities. We detected a 1602 bp cDNA fragment of GALNTL5 that included a 1326 bp coding sequence, encoding 441 amino acids and 90 and 185 bp of the 5' and 3' untranslated regions, respectively. The GALNTL5 amino acid sequence showed 51.87-83.48% identity with the sequences of proteins from other species. It was detected exclusively in the testis and the levels of both the mRNA and protein were progressively increased with age. Immunohistochemistry further revealed that GALNTL5 specifically localized in the elongating spermatids and spermatozoa, and it was demonstrated to be strongly concentrated in the head, neck, and mid-piece region of spermatozoa by immunocytochemistry. The sperm density and the percentage of live sperm in the high motility group (≥80%) were significantly higher than in the low motility group (≤50%), and the reverse trend was observed with the abnormal sperm. Western blot analysis showed that the protein expression of PGK2, ALDOA, and GALNTL5 were significantly higher in the high motility group than in the low motility group. Overall, the data suggest that GALNTL5 is an important functional molecule during spermatogenesis. Moreover, it is the first to suggest that the expression level of GALNTL5 is positively correlated with the sperm motility.

Yao X ，Yang H ，Zhang Y ，Ren C ，Nie H ，Fan Y ，Zhou W ，Wang S ，Feng X ，Wang F ... -  《-》

A heterozygous mutation of GALNTL5 affects male infertility with impairment of sperm motility.

Takasaki N ，Tachibana K ，Ogasawara S ，Matsuzaki H ，Hagiuda J ，Ishikawa H ，Mochida K ，Inoue K ，Ogonuki N ，Ogura A ，Noce T ，Ito C ，Toshimori K ，Narimatsu H ... -  《-》

Expression and localization of polypeptide N-acetylgalactosaminyltransferase-like protein 5 in the reproductive organs and sperm of Hu sheep.

The polypeptide N-acetylgalactosaminyltransferase-like protein 5 (GALNTL5) is involved in male fertility; however, its involvement in the reproduction and fertility of females remains unclear. Therefore, the present study aimed to examine the presence of GALNTL5 in the reproductive organs of ewes during the estrus period and to investigate its expression in cauda epididymal and ejaculated sperm. Results showed that GALNTL5 mRNA and protein were present in some reproductive organs of ewes during the estrus period. The highest levels of GALNTL5 mRNA and protein occurred in the uterine horn and oviductal ampulla and the lowest in the uterine cervix and oviductal infundibulum. Immunohistochemical analysis revealed that GALNTL5 protein was mainly located in luminal and glandular epithelial cells of the uterus and oviduct, and in the theca and granulosa cells of the ovary. GALNTL5 gene expression was significantly higher in ejaculated sperm than in cauda epididymal sperm. The amount of GALNTL5 protein in seminal plasma was significantly higher than in ejaculated sperm. Additionally, GALNTL5 was strongly localized in the mid-piece and head of ejaculated sperm, and in the head-tail coupling apparatus and acrosome of cauda epididymal sperm. This is the first evidence that GALNTL5 might play an important role in a range of reproductive functions as well as in sperm motility and capacitation. Further studies are required to evaluate the function of GALNTL5 in reproduction.

Yao X ，Ei-Samahy MA ，Feng X ，Zhang T ，Li F ，Zhang G ，Pang J ，Nie H ，Wang F ... -  《-》

A novel splice variant of the bovine GALNTL5 gene identified in Chinese Holstein bull testis tissue and its mRNA expression.

Li H ，Jiang Q ，Ju ZH ，Huang JM ，Wang XG ，Yang CH ，Sun Y ，Yang GW ，Zhong JF ，Wang CF ... -  《-》

The expression characteristics of FAM71D and its association with sperm motility.

What are the features of FAM71D (Family with sequence similarity 71, member D) expression and is there an association between FAM71D expression and sperm motility? FAM71D, a novel protein exclusively expressed in the testis, is located in sperm flagella and is functionally involved in sperm motility. Some testis-specific proteins have been reported as potential diagnostic biomarkers to evaluate the spermatogenesis process and sperm quality. We have identified a novel testis-specific protein, FAM71D, through microarray data analysis, yet little is known about its expression and function. FAM71D mRNA and protein expression was quantified during mouse testis development. Its localization in germ cells was detected by dual-labeled immunostaining in testis sections and sperm smears. The clinical significance was assessed by comparing FAM71D expression in spermatozoa from normozoospermic controls and asthenozoospermic patients. Testes were dissected from C57BL/6 J male mice at postnatal ages of 1, 2, 3, 4, 6, 8 weeks and 6 months, and sperm was collected from cauda epididymides of adult mice by the swim-up method. Human spermatozoa were isolated from 100 human semen samples by density gradient Percoll centrifugation. RT-qPCR and western blot were performed to semi-quantify the expression of FAM71D in mouse testis, and in the ejaculated spermatozoa of normozoospermic controls and asthenozoospermic patients. Immunofluorescence staining was used to detect the localization of FAM71D. Co-immunoprecipitation assay was performed to evaluate the interaction between FAM71D and calmodulin. An antibody blocking assay was employed to assess the role of FAM71D in sperm motility. Our results showed that FAM71D was exclusively expressed in the testis in an age-dependent manner. FAM71D expression exhibited dynamic change in the cytoplasm of spermatids during spermiogenesis and was finally retained in sperm flagella. FAM71D could interact with calmodulin. Use of anti-FAM71D antibody on sperm significantly decreased sperm motility. Expression level of FAM71D was markedly reduced in the ejaculated spermataozoa of asthenozoospermic patients (P < 0.05), and this was correlated with sperm progressive motility (r = 0.7435, P < 0.0001). N/A. The sample size was limited and it is necessary to verify the correlation of FAM71D expression with sperm motility in larger cohorts. Furthermore, our results were descriptive and follow-up studies would be needed to elucidate the detailed role of FAM71D in sperm motility. This is the first systematic study to document the expression of endogenous FAM71D and a function for FAM71D in sperm motility. It provides new insights into our understanding of sperm motility regulation and causes of male infertility. This study was funded by the National Natural Science Foundation of China, Guangdong Natural Science Foundation and the Shenzhen Project of Science and Technology. The authors have no competing interests.

Ma Q ，Li Y ，Luo M ，Guo H ，Lin S ，Chen J ，Du Y ，Jiang Z ，Gui Y ... -  《-》