Reproductive toxicity of 1-bromopropane, a newly introduced alternative to ozone layer depleting solvents, in male rats.

1-Bromopropane has been newly introduced as an alternative to ozone-depleting solvents. We aimed to clarify its dose-dependent reproductive toxicity in male rats. Thirty-six Wistar male rats were randomly divided into 4 groups of 9. The groups were exposed to 200, 400, or 800 ppm 1-bromopropane or only fresh air, 8 h per day for 12 weeks. Epididymal sperm indices were evaluated after a 12-week exposure. The testes, epididymides, seminal vesicle, prostate, and other organs were weighed and examined histopathologically. Spermatogenic cells, in stage VII seminiferous tubules, and retained spermatids, at the basal region of stages IX-XI seminiferous epithelium, were counted. Plasma testosterone levels were measured by radioimmunoassay. The testicular weight did not significantly change, but the weight of epididymides, seminal vesicle, and prostate dose-dependently decreased. The weight of seminal vesicle decreased significantly at the lowest concentration of 200-ppm and over. 1-Bromopropane induced a dose-dependent decrease in the epididymal sperm count and in motility, as well as an increase in tailless sperm and sperm with an immature head shape. The spermatogonia, preleptotene spermatocytes, pachytene spermatocytes, and round spermatids did not decrease significantly at stage VII. Retained, elongated spermatids near the basement membrane at the postspermiation stages IX-XI increased dose-dependently. Plasma testosterone levels significantly decreased at the 800-ppm dosage. 1-Bromopropane caused failure of spermiation. Its reproductive toxicity is different from that of 2-bromopropane, which specifically impairs spermatogonia. Thus, this solvent may have serious reproductive toxic effects in men, and should be used very cautiously in the workplace.     

Unique alternative translation from two open reading frames on Acpin1 mRNA yields an acrosomal protein and a salivary-gland-specific protein

Objective:   To examine the expression profiles of the proteins translated from Acpin1 mRNA in germ cells.
Methods:   Northern and western blotting of various tissues and immunohistochemical analysis of germ cells were carried out in a mouse model.
Results:   ACPIN1 protein was transcribed from the longer, 3' open reading frame (ORF) of Acpin1. An alternative-splicing variant, Acpin1vs , contained only the smaller, 5' ORF of the full-length Acpin1 gene. Its gene product, SAGSIN1, was expressed specifically in salivary glands. Retrotransposed regions of Acpin1 homology were also detected in various chromosomes, and intronless paralogous genes on the X chromosome were expressed in the testis and other tissues. The genomic structure of Acpin1 is highly conserved in mammals.
Conclusion:   The two ORFs on the Acpin1 mRNA are independently translated in differentiated cells. Analysis of gene Acpin1 might clarify the molecular mechanism of spermatogenesis.     

Genetic loss or pharmacological blockade of testes-expressed taste genes causes male sterility

TAS1R taste receptors and their associated heterotrimeric G protein gustducin are involved in sugar and amino acid sensing in taste cells and in the gastrointestinal tract. They are also strongly expressed in testis and sperm, but their functions in these tissues were previously unknown. Using mouse models, we show that the genetic absence of both TAS1R3, a component of sweet and amino acid taste receptors, and the gustducin α-subunit GNAT3 leads to male-specific sterility. To gain further insight into this effect, we generated a mouse model that expressed a humanized form of TAS1R3 susceptible to inhibition by the antilipid medication clofibrate. Sperm formation in animals without functional TAS1R3 and GNAT3 is compromised, with malformed and immotile sperm. Furthermore, clofibrate inhibition of humanized TAS1R3 in the genetic background of Tas1r3^−/−, Gnat3^−/− doubly null mice led to inducible male sterility. These results indicate a crucial role for these extraoral “taste” molecules in sperm development and maturation. We previously reported that blocking of human TAS1R3, but not mouse TAS1R3, can be achieved by common medications or chemicals in the environment. We hypothesize that even low levels of these compounds can lower sperm count and negatively affect human male fertility, which common mouse toxicology assays would not reveal. Conversely, we speculate that TAS1R3 and GNAT3 activators may help infertile men, particularly those that are affected by some of the mentioned inhibitors and/or are diagnosed with idiopathic infertility involving signaling pathway of these receptors.     

Asthenozoospermia: Cellular and molecular contributing factors and treatment strategies

Semen sample with poor sperm motility, which called asthenozoospermia, is considered as one of the main factors contributing to male infertility. Recognition of the cellular and molecular pathways contributing to sperm motility reduction may lead to applying novel treatment strategies for overcoming low sperm motility in asthenozoospermia individuals. In this review, we intend to discuss the main causes of sperm motility reduction in asthenozoospermia and some treatment strategies used to overcome low sperm motility.     

Electron microscopy analysis of histone acetylation and DNA strand breaks in mouse elongating spermatids using a dual labelling approach

Chromatin remodelling steps in mammalian spermatids include post‐translational modifications of histones and DNA fragmentation. Histone H4 hyperacetylation (AcH4) establishes a chromatin state that facilitates DNA repair in somatic cells. So we sought to determine whether a similar link exists in spermatids by combining immunogold labelling with detection of DNA strand breaks, making use of gold particles of different sizes. DNA strand breaks were not detected in the vicinity of AcH4 chromatin, suggesting that this modified histone may not be involved in the aetiology of DNA fragmentation and repair in spermatids. The AcH4 reactivity, however, indicates that chromatin remodelling is distributed throughout the nucleus.     

Ultrastructure of spermatogenesis of <Emphasis Type="Italic">Taenia taeniaeformis</Emphasis> (Batsch, 1786) (Cestoda,Cyclophyllidea, Taeniidae) and comparison of spermatological characters in the family Taeniidae Ludwig, 1886

The ultrastructure of spermatogenesis of Taenia taeniaeformis is described for the first time by means of transmission electron microscopy (TEM). Mature testes contain all stages of spermatogenesis; primary spermatogonia are usually situated at the periphery and mature spermatozoa in the centre of testes. The general process is similar to that described in other cestodes. Six incomplete, synchronic cytokineses occur: four mitotic and two meiotic cell divisions. All these divisions occur simultaneously, resulting in a rosette cluster of four tertiary spermatogonia, then eight quaternary spermatogonia, and subsequently sixteen primary spermatocytes. All of these enter into a growth period and their enlarged nuclei move to the periphery of cells of the rosettes. The first meiotic division forms thirty-two secondary spermatocytes and after the second meiotic division, there are sixty-four spermatids. Spermiogenesis in T. taeniaeformis corresponds to the Ba and Marchand’s Type 3 and begins with the formation of a differentiation zone in the form of a conical projection of cytoplasm delimited by a ring of arching membranes and surrounded by submembranous cortical microtubules. Within this area, there are two centrioles, orthogonally disposed, and vestigial striated rootlets. Only one of the centrioles develops a flagellum that grows externally to the cytoplasmic extension. Posteriorly, a flagellar rotation inferior to 90° occurs and the flagellum becomes parallel to the cytoplasmic extension. Later, the two processes fuse during the so-called proximodistal fusion. The nucleus elongates and moves into the cytoplasmic extension. In the final stage of spermiogenesis, a single crested body appears at the base of the differentiating spermatozoon. Finally, the ring of arching membranes constricts and the young spermatozoon detaches from the residual cytoplasm. Ultrastructural aspects of spermatogenesis are compared with that of other cestodes studied to date, particularly of the family Taeniidae. Dedicated to the memory of Professor Krystyna Rybicka on the occasion of 85th Anniversary of her birthday     

Immunolocalization assessment of metastasis-associated protein 1 in human and mouse mature testes and its association with spermatogenesis

Aim： To investigate the stage-specific localization of metastasis-associated protein 1 （MTA1） during spermatogenesis in adult human and mouse testis. Methods： The immunolocalization of MTA1 was studied by immunohistochemistry and Western blot analysis. The distribution pattern of MTA1 in mouse testis was confirmed by using quantitative analysis of purified spermatogenic cells. Results： The specificity of polyclonal antibody was confirmed by Western blot analysis. MTA1 was found expressed in the nucleus of germ cells, except elongate spermatids, and in the cytoplasm of Sertoli cells; Leydig cells did not show any specific reactivity. MTA1 possessed different distribution patterns in the two species： in humans, the most intensive staining was found in the nucleus of round spermatids and of primary spermatocytes while in mice, the most intense MTA 1 staining was in the nucleus of leptotene, zygotene and pachytene spermatocytes. In both species the staining exhibited a cyclic pattern. Conclusion： The present communication initially provides new evidence for the potential role of MTA1 in mature testis. In addition, its distinctive expression in germ cells suggests a regulatory role of the peptide during spermatogenesis.     

Expression of Mammalian P1 Protamine Gene

Expression of Mammalian P1 Protamine Gene