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Comparative study of the clastogenic efficiency of ethyl methanesulfonate and diethyl sulfate in Drosophila melanogaster mature sperm.

Chromosome loss and translocation tests were carried out in Drosophila melanogaster sperm, stored in untreated females for up to 24 days, to compare the clastogenicity of ethyl methanesulfonate (EMS) and diethyl sulfate (DES). The sex-linked recessive lethal test was used as a "biological dosimeter" and the following results were obtained: The yield of 2-3 translocations induced by both mutagens increased steadily with storage, being significantly higher after EMS than after DES treatment. The frequencies of partial losses induced by EMS and DES were similar and increased with storage. With up to 11 days' storage, the frequency of complete loss induced by DES was higher than that induced by EMS and remained unchanged when storage was extended to 24 days. Complete loss induced by EMS increased significantly with further storage (12-24 days). With DES, complete (but not partial) loss was detected with a dose at which EMS failed to modify the control values. These data suggest that the lower recovery of II-III translocations after treatment with DES does not result from a low breaking capacity but from a diminished or delayed rejoining of the induced breaks. This could be due to a physiological impairment of the treated cells by the high toxicity of DES or to an actual lower rejoinability of the broken ends. The differential recovery of complete and partial losses after DES treatment further suggests that the mechanisms leading to the fixation of both types of damage are somehow different, and that processes intervening in the recovery of partial losses are less affected, or not at all, by the proposed reduced rejoining of chromosome breaks.

Aneuploidy↗

Ectopic epididymal expression of guinea pig intestinal phospholipase B. Possible role in sperm maturation and activation by limited proteolytic digestion.

Guinea pig intestinal phospholipase B is a calcium-independent phospholipase hydrolyzing sequentially the acyl ester bonds at sn-2 and sn-1 positions of glycerophospholipids, promoting the formation of sn-glycero-3-phosphocholine from phosphatidylcholine. This 140-kDa glycoprotein from the brush border membrane of differentiated enterocytes contributes to lipid digestion as an ectoenzyme. The cDNA coding for guinea pig phospholipase B was revealed to be the homologue of AdRab-B, an mRNA appearing in rabbit upon intestine development. The sequence predicts a polypeptide of 1463 amino acids displaying four homologous repeats, two of them containing the lipase consensus sequence GXSXG. A 5-kilobase transcript was particularly abundant in mature ileal and jejunal enterocytes but was also detected in epididymis, where phospholipase B displayed a higher molecular mass (170 kDa versus 140 kDa in intestine), with no obvious evidence for enzyme activity. Trypsin treatment of phospholipase B immunoprecipitated from epididymal membranes reduced its size to 140 kDa, coinciding with the appearance of a significant phospholipase A2 activity. The same results were obtained in COS cells transfected with phospholipase B cDNA. Since sn-glycero-3-phosphocholine present at high concentrations in seminal plasma mainly stems from epididymis, this suggests a possible role of phospholipase B in male reproduction. This novel localization also unravels a mechanism of phospholipase B activation by limited proteolysis involving either trypsin in the intestinal lumen or a trypsin-like endopeptidase in the male reproductive tract.

Amino Acid Sequence↗

Silent cells? Potential for context-dependent gene expression in mature sperm.

Sperm are traditionally viewed as transcriptionally and translationally silent cells. However, observations that components of the cellular machinery of gene expression are maintained in ejaculated sperm are increasingly cited as challenges to this fundamental assumption. Here, we critically evaluate these arguments and present three lines of evidence from both model and non-model systems that collectively raise the question of whether ejaculated sperm may be capable of active gene expression. First, and critical for arguments surrounding the possibility of differential gene expression, we review recent evidence that spermatozoa may retain the capacity to transcribe and translate their genomes. Second, we highlight how sperm cells can exhibit differential transcript quantities across different post-ejaculation environments. Third, we ask whether the accumulating evidence of remarkable phenotypic plasticity in post-ejaculatory sperm phenotypes could be mechanistically underpinned by changes in sperm gene expression. While these lines of evidence are indirect and do not definitively show transcription of sperm genomes, we highlight how emerging technologies may enable us to test this hypothesis explicitly. Our review advocates for progress in this field and highlights several important evolutionary, ecological and practical implications that will probably transcend disciplines to the clinical and applied reproductive sectors.

Male↗

Microfluorometric assessment of sperm maturation in testicular biopsies from men with histologically normal or reduced spermatogenesis.

Spermatogenesis has been studied in testicular biopsies by means of microfluorometric assessment of the DNA-DNP complex using ethidium bromide in 10 men with histologically normal spermatogenesis and in 10 men with histologically reduced spermatogenesis. The expected haploid DNA value was found in the round spermatids, whereas only 70% of the haploid fluorescence value was found in the elongated spermatids and around 60% in the testicular spermatozoa. No difference was found in the mean fluorescence values between the pathological group and the controls. it is suggested that structural changes or an increase in basic nuclear proteins gradually exclude the ethidium bromide from the binding to the DNA molecule and that this phenomenon occurs concomitantly with a decrease in double-stranded RNA. The difference in fluorescence intensity before and after RNAase treatment was regarded as an approximate estimate of the RNA content. There was a significant decrease of these values during spermatogenesis.

Adult↗