Genetics of Male and Female Sterility in Hybrids of Drosophila pseudoobscura and D. persimilis

The genetic basis of male and female sterility in hybrids of Drosophila pseudoobscura-Drosophila persimilis was studied using backcross analysis. Previous studies indirectly assessed male fertility by measuring testis size; these studies concluded that male sterility results from an X chromosome-autosome imbalance. By directly scoring for the production of motile sperm, male sterility is shown to be largely due to an incompatibility between genes on the X and Y chromosomes of these two species. These species have diverged at a minimum of nine loci affecting hybrid male fertility. Semisterility of hybrid females appears to result from an X chromosome-cytoplasm interaction; the X chromosome thus has the largest effect on sterility in both male and female hybrids. This is apparently the first analysis of the genetic basis of female sterility, or of sterility/inviability affecting both sexes, in an animal hybridization.     

The effects of inbreeding and heat stress on male sterility in Drosophila melanogaster

Understanding the consequences of inbreeding in combination with stress is important for the persistence of small endangered populations in a changing environment. Inbreeding and stress can influence the population at all stages of the life cycle, and in the last two decades a number of studies have demonstrated inbreeding depression for most life‐cycle components, both in laboratory populations and in the wild. Although male fertility is known to be sensitive to temperature extremes, few studies have focused on this life‐cycle component. We studied the effects of inbreeding on male sterility in benign and stressful environments using Drosophila melanogaster as a model organism. Male sterility was compared in 21 inbred lines and five non‐inbred control lines at 25.0 and 29.0 °C. The effect of inbreeding on sterility was significant only at 29.0 °C. This stress‐induced increase in sterility indicates an interaction between the effects of inbreeding and high‐temperature stress on male sterility. In addition, the stress‐induced temporary and permanent sterility showed significant positive correlation, as did stress‐induced sterility and the decrease in egg‐to‐adult viability. This suggests that the observed stress‐induced decline in fitness could result from conditionally expressed, recessive deleterious alleles affecting both sterility and viability simultaneously. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 432–442.     

Misregulation of spermatogenesis genes in Drosophila hybrids is lineage‐specific and driven by the combined effects of sterility and fast male regulatory divergence

Hybrid male sterility is a common outcome of crosses between different species. Gene expression studies have found that a number of spermatogenesis genes are differentially expressed in sterile hybrid males, compared with parental species. Late‐stage sperm development genes are particularly likely to be misexpressed, with fewer early‐stage genes affected. Thus, a link has been posited between misexpression and sterility. A more recent alternative explanation for hybrid gene misexpression has been that it is independent of sterility and driven by divergent evolution of male‐specific regulatory elements between species (faster male hypothesis). The faster male hypothesis predicts that misregulation of spermatogenesis genes should be independent of sterility and approximately the same in both hybrids, whereas sterility should only affect gene expression in sterile hybrids. To test the faster male hypothesis vs. the effect of sterility on gene misexpression, we analyse spermatogenesis gene expression in different species pairs of the Drosophila phylogeny, where hybrid male sterility occurs in only one direction of the interspecies cross (i.e. unidirectional sterility). We find significant differences among genes in misexpression with effects that are lineage‐specific and caused by sterility or fast male regulatory divergence.     

Reproductive isolation caused by azoospermia in sterile male hybrids of Drosophila

Recently diverged populations in the early stages of speciation offer an opportunity to understand mechanisms of isolation and their relative contributions. Drosophila willistoni is a tropical species with broad distribution from Argentina to the southern United States, including the Caribbean islands. A postzygotic barrier between northern populations (North America, Central America, and the northern Caribbean islands) and southern populations (South American and the southern Caribbean islands) has been recently documented and used to propose the existence of two different subspecies. Here, we identify premating isolation between populations regardless of their subspecies status. We find no evidence of postmating prezygotic isolation and proceeded to characterize hybrid male sterility between the subspecies. Sterile male hybrids transfer an ejaculate that is devoid of sperm but causes elongation and expansion of the female uterus. In sterile male hybrids, bulging of the seminal vesicle appears to impede the movement of the sperm toward the sperm pump, where sperm normally mixes with accessory gland products. Our results highlight a unique form of hybrid male sterility in Drosophila that is driven by a mechanical impediment to transfer sperm rather than by an abnormality of the sperm itself. Interestingly, this form of sterility is reminiscent of a form of infertility (azoospermia) that is caused by lack of sperm in the semen due to blockages that impede the sperm from reaching the ejaculate.     

The role of the microbial factor in the pathogenesis of male infertility

Microorganisms can be isolated from most seminal fluid samples, but the significance of bacteriospermia is uncertain because many males lack symptoms associated with the bacterial infection of the reproductive tract. The data on the influence of urogenital tract infections of fertility are contradictory. In many cases opportunistic microorganisms cause such classical infections of the urogenital tract as epididymitis and prostatitis, as well as subclinical reproductive tract infections. Some possible pathophysiological mechanisms of the development of infertility linked with infection of the ejaculate are considered: its direct effect on the fertile properties of the seminal fluid due to a decrease in the number of spermatozoa, the suppression of their motility, changes in their morphology and fertilizing capacity, its indirect influence due to the inhibition of spermatogenesis resulting from testicular damage, autoimmune processes induced by inflammation, secretory dysfunction of the male accessory sex glands as a consequence of the infection of the reproductive tract organs, leukocytospermia with its secondary influence of the ejaculate parameters, etc. The importance of the pathogenic properties of microorganisms for the localization of the inflammatory process in the urogenital tract of males and their role in the pathogenesis of male infertility are discussed.     

The mouse t complex distorter/sterility candidate, Dnahc8, expresses a gamma-type axonemal dynein heavy chain isoform confined to the principal piece of the sperm tail

Heterozygosity for a t haplotype (t) in male mice results in distorted transmission (TRD) of the t-bearing chromosome 17 homolog to their offspring. However, homozygosity for t causes male sterility, thus limiting the spread of t through the population at large. The Ca(2+)-dependent sperm tail curvature phenotypes, "fishhook", where abnormally high levels of sperm exhibit sharp bends in the midpiece, and "curlicue", where motile sperm exhibit a chronic negative curving of the entire tail, have been tightly linked to t-associated male TRD and sterility traits, respectively. Genetic studies have indicated that homozygosity for the t allele of Dnahc8, an axonemal gamma-type dynein heavy chain (gammaDHC) gene, is partially responsible for expression of "curlicue"; however, its involvement in "fishhook"/TRD, if any, is unknown. Here we report that the major isoform of DNAHC8 is copiously expressed, carries an extended N-terminus and full-length C-terminus, and is stable and equally abundant in both testis and sperm from +/+ and t/t animals. By in silico analysis we also demonstrate that at least three of the seventeen DNAHC8(t) mutations at highly conserved positions in wild-type DHCs may be capable of substantially altering normal DNAHC8 function. Interestingly, DNAHC8 is confined to the principal piece of the sperm tail. The combined results of this study suggest possible mechanisms of DNAHC8(t) dysfunction and involvement in "curlicue", and support the hypothesis that "curlicue" is a multigenic phenomenon. They also demonstrate that the accelerated "fishhook" phenotype of sperm from +/t males is not directly linked to DNAHC8(t) dysfunction.     

Fécondation assistée par micro injection des spermatides: Etat des lieux et questions posées

In order to treat male infertility, since 5 years ago, intracytoplasmic sperm injection (ICSI) represents a new therapeutic approach of different forms of male sterility including obstructive and spermatogenic failure azoospermia. This paper reviews the use of ICSI with round or elongated spermatids.     

Genes in the first and fourth inversions of the mouse t complex synergistically mediate sperm capacitation and interactions with the oocyte

The t haplotypes (t) are recent evolutionary derivatives of an alternate form of the mouse t complex region located at the proximal end of chromosome 17. This variant form of approximately 1% of the mouse genome is a source of mutations altering numerous sperm functions crucial for fertilization. Males that carry two t haplotypes (t/t) are invariably sterile. t haplotypes contain four inversions relative to the wild-type t complex (+), so that in matings involving a +/t heterozygote, t is usually transmitted as a single unit. However, rare recombinants have been recovered, which carry only part of the t genotype and express only some of the t-dependent phenotypes. Use of these partial t haplotypes in genetic crosses has resulted in the general location of the two major t male sterility factors, S1 and S2, within inversions 1 and 4, respectively. Since sterility can result from a plethora of sperm defects, we have made a detailed study of various functional parameters of sperm from mice carrying S1 or S2 heterozygously or homozygously or in combination. Both S1 and S2 contain mutations altering sperm functions, including motility, capacitation, binding to the zona pellucida, binding to the oocyte membrane, and penetration of the zona pellucida-free oocyte. Therefore it seems clear that each of these factors contains multiple genes contributing to sterility. Furthermore, our results indicate that genes within S1 interact with genes in S2 for all sperm functions examined. However, S1 and S2 genes affecting motility interact in a purely additive fashion, while S1 and S2 genes affecting most other sperm characteristics interact in a synergistic manner. Additionally, the patterns of synergism between S1 and S2 for abnormalities in capacitation, sperm-oolemma binding, and zona-free oocyte penetration are nearly identical. This suggests that these three defects are caused by mutation of the same gene within each sterility factor. These findings will not only be instrumental in matching the various t haplotype sperm defects to candidate genes for S1 and S2, but will facilitate a more comprehensive understanding of the cellular and genetic mechanisms underlying t haplotype male sterility.     

The contribution of the y chromosome to hybrid male sterility in house mice

Hybrid sterility in the heterogametic sex is a common feature of speciation in animals. In house mice, the contribution of the Mus musculus musculus X chromosome to hybrid male sterility is large. It is not known, however, whether F(1) male sterility is caused by X-Y or X-autosome incompatibilities or a combination of both. We investigated the contribution of the M. musculus domesticus Y chromosome to hybrid male sterility in a cross between wild-derived strains in which males with a M. m. musculus X chromosome and M. m. domesticus Y chromosome are partially sterile, while males from the reciprocal cross are reproductively normal. We used eight X introgression lines to combine different X chromosome genotypes with different Y chromosomes on an F(1) autosomal background, and we measured a suite of male reproductive traits. Reproductive deficits were observed in most F(1) males, regardless of Y chromosome genotype. Nonetheless, we found evidence for a negative interaction between the M. m. domesticus Y and an interval on the M. m. musculus X that resulted in abnormal sperm morphology. Therefore, although F(1) male sterility appears to be caused mainly by X-autosome incompatibilities, X-Y incompatibilities contribute to some aspects of sterility.     

Mutation of cyp19a1b results in sterile males due to efferent duct obstruction in Nile tilapia

The involvement of estrogen in male fertility has been well established in mammals. However, less is known about the role of estrogen in fish male reproduction. Our recent study revealed that Cyp19a1a deficiency had no effect on fertility in male fish. In this study, expression of Cyp19a1b, but not Cyp19a1a, was detected by immunohistochemistry in Leydig cells of tilapia testes. cyp19a1b mutation resulted in a significant decrease in the concentration of 17β‐estradiol in serum and sterility in XY fish, as no offspring were obtained when crossed with control XX fish at 240 days after hatching (dah). No sperm was obtained from the mature mutants by in vitro extrusion. Further examination of the mutant gonads revealed excessive semen accumulation and testicular hypertrophy. Semen collected from the mutant testes during autopsy contained sperm with a normal morphology that showed no significant differences in motility, VCL, BCF, STR, or fertility compared with control sperm. Efferent ducts from the mutant testes, which had low‐convolution levels, fewer branches, and no blood vessels observed inside the walls, were significantly smaller in size. qRT‐PCR analyses showed downregulated expression of ion exchange genes. There was increased apoptosis in the epithelial cells of the efferent ducts and other somatic cells of the testes as revealed by TUNEL staining, as well as upregulation of apoptosis gene expression in the mutants. At 360 dah, mutant fish showed testicular atrophy and efferent duct fibrosis. These results demonstrated that estrogen deficiency caused by Cyp19a1b mutation resulted in male sterility due to efferent duct obstruction.     

Effects on spermiogenesis in the mouse of a male sterile neurological mutation,Purkinje cell degeneration

Purkinje cell degeneration (pcd) is a neurological mutation in the mouse that causes male sterility, but not female sterility. In order to assess the effects of this mutation on spermiogenesis, the structure of the testis and of epididymal spermatozoa was examined by transmission and scanning electron microscopy. In the mutant males, the sperm count was reduced, sperm were nonmotile, and 93% of the sperm were characterized by structural abnormalities of the head, the tail, or both. In the testes of mutant mice, Sertoli cell structure was normal, as were also the early stages of spermiogenesis. However, the elongating and maturing spermatids were characterized by abnormally shaped heads and tails with extraneous and ectopic outer dense fibers. These defects were common in the testes of the mutant mice and rare in the testes of the littermate control mice. It was concluded that the structural abnormalities of the pcd sperm occurred during spermiogenesis and were not due to degeneration of the sperm in the epididymis. These structural abnormalities are similar to those found in all other reported male sterile mutants of the mouse; therefore, although they are caused by the expression of the pcd gene, they are not unique to the expression of this gene.     

Haldane’s Rule Is Linked to Extraordinary Sex Ratios and Sperm Length in Stalk-Eyed Flies

We use three allopatric populations of the stalk-eyed fly Teleopsis dalmanni from Southeast Asia to test two predictions made by the sex chromosome drive hypothesis for Haldane’s rule. The first is that modifiers that suppress or enhance drive should evolve rapidly and independently in isolated populations. The second is that drive loci or modifiers should also cause sterility in hybrid males. We tested these predictions by assaying the fertility of 2066 males derived from backcross experiments involving two pairs of populations and found that the proportion of mated males that fail to produce any offspring ranged from 38 to 60% among crosses with some males producing strongly female-biased or male-biased sex ratios. After genotyping each male at 25–28 genetic markers we found quantitative trait loci (QTL) that jointly influence male sterility, sperm length, and biased progeny sex ratios in each pair of populations, but almost no shared QTL between population crosses. We also discovered that the extant X^SR chromosome has no effect on sex ratio or sterility in these backcross males. Whether shared QTL are caused by linkage or pleiotropy requires additional study. Nevertheless, these results indicate the presence of a “cryptic” drive system that is currently masked by suppressing elements that are associated with sterility and sperm length within but not between populations and, therefore, must have evolved since the populations became isolated, i.e., in <100,000 years. We discuss how genes that influence sperm length may contribute to hybrid sterility.     

Double copulation of a female with sterile diploid and polyploid males recovers fertility in Bombyx mori

Silkworm males produce dimorphic sperm, nucleate eupyrene sperm and anucleate apyrene sperm. Apyrene sperm have been speculated to have an assisting role in fertilisation. However, the coexistence of eupyrene and apyrene sperm in the testis and female reproductive organs has made it difficult to define the role of apyrene sperm. Polyploid males are highly sterile. Microscopic observation revealed that the elimination of eupyrene nuclei by peristaltic squeezing caused the sterility of polyploids. Heat-shock applied to pupae of Daizo males (DH) also induced high sterility due to the lack of normal apyrene sperm. When eupyrene sperm of sterile DH males and apyrene sperm of sterile polyploid males were mixed by double copulation, a remarkable increase in fertility of the double-mated females was observed. This finding strongly suggests that the apyrene sperm are indispensable in fertilisation of the silkworm and that polyploid apyrene sperm function as a substitute for diploid sperm. We established an experimental system in which we can separate the two types of sperm for further studies on their functions without chemical and/or mechanical treatments.     

Hidden effects of X chromosome introgressions on spermatogenesis in Drosophila simulans x D. mauritiana hybrids unveiled by interactions among minor genetic factors.

One of the most frequent outcomes of interspecific hybridizations in Drosophila is hybrid male sterility. Genetic dissection of this reproductive barrier has revealed that the number of responsible factors is very high and that these factors are frequently engaged in complex epistatic interactions. Traditionally, research strategies have been based on contrasting introgressions of chromosome segments that produce male sterility with those that allow fertility. Few studies have investigated the phenotypes associated with the boundary between fertility and sterility. In this study, we cointrogressed three different X chromosome segments from Drosophila mauritiana into D. simulans. Hybrid males with these three segments are usually fertile, by conventional fertility assays. However, their spermatogenesis shows a significant slowdown, most manifest at lower temperatures. Each of the three introgressed segments retards the arrival of sperm to the seminal vesicles. Other small disturbances in spermatogenesis are evident, which altogether lead to an overall reduction in the amount of motile sperm in their seminal vesicles. These results suggest that a delay in the timing of spermatogenesis, which might be brought about by the cumulative action of many different factors of minor segment, may be the primary cause of hybrid male sterility.     

Male Sterility in Tomato

Male sterility research has been directed toward two goals: identifying genes required for the pollen development pathway and, more practically, identifying genetically stable lines that can be used in hybrid seed-breeding programs. The present resurgence of interest in male sterility remains true to these goals, but in addition seeks a molecular understanding of pollen development in order to genetically engineer controllable male sterility for hybrid seed production. In this review, we discuss the genetic and histochemical studies of tomato male sterile mutants that were conducted prior to 1970 in the context of gene expression and interaction. We also examine the use of molecular biological techniques in recent studies of male sterility and report on the current strategies being used for hybrid seed production.     

The interaction between X-rays and transposon mobility in Drosophila: hybrid sterility and chromosome loss

Genetic traits associated with P-M hybrid dysgenesis in Drosophila melanogaster were synergistically affected by X-rays. The interaction between damages induced by these two mutator systems was evident when sterility and X/Y chromosome loss were used as endpoints. No interaction was detected in partial chromosome loss, monitored by the loss of BS and y+ markers. The synergism in sterility, measured either as all-or-none or premature sterility (fecundity) was observed when male hybrids derived from different P strains fathers, namely Harwich or II2, were X-irradiated and the effects compared relative to similarly treated non-dysgenic hybrids. Brooding of sperm showed that the effects of ionizing radiation were ionizing radiation were dependent upon the stage of spermatogenesis during which cells were irradiated. The highly synergistic effect on sterility was found when either spermatids or spermatocytes, but not mature sperm, were irradiated with 550 rad of X-rays. These findings were consistent with the higher radiosensitivity of spermatocytes and spermatids to genetic damage and with the correlation between the incidence of sterility and aging of dysgenic hybrids. The latter observation was particularly evident in the case of Harwich P strain-derived male hybrids whose fertility was greatly reduced due to P element mobility. The synergistic effect of X-rays in these dysgenic hybrids resulted in the virtual abolition of the germ line, increasing the sterility from 50% of the untreated 9-10-day old males, to 85% of the treated males when spermatocytes were irradiated. The synergism observed between transposon mobility and ionizing radiation can best the attributed to an interaction between X-ray-induced and P element-induced chromosome breaks. This interpretation is consistent with the more than additive increase in X or Y chromosome loss in irradiated, Harwich P strain-derived hybrid sons. The induction of these events was 1.164% in dysgenic irradiated males as compared to 0.234% in X-irradiated nondysgenic hybrids and 0.40% in dysgenic untreated males. No synergism was observed in X/Y loss in hybrids derived from II2 P strain fathers where the frequency of the events due to P element mobility alone was only one tenth (0.037%) of that found in Harwich-derived hybrids.     

Drosophila bipectinata species complex: study of phylogenetic relationship among four members through the analysis of morphology of testes and seminal vesicles

Species maintain their identity through reproductive isolating mechanisms, which are broadly classified into prezygotic and postzygotic isolating mechanisms. In the Drosophila bipectinata species complex, investigations were made on the degree of crossability (a prezygotic isolating mechanism) and the causes of hybrid male sterility (a postzygotic isolating mechanism) to analyse the phylogenetic relationship. Among the four species, D. bipectinata crosses with Drosophila parabipectinata freely in one direction and both of them also cross with Drosophila malerkotliana easily but it is difficult to cross all the three species with Drosophila pseudoananassae . In the hybrids involving D. pseudoananassae , no sperm were observed indicating high degree of perturbance during spermatogenesis while in the other hybrids immotile sperm were present indicating comparatively less disturbance during spermatogenesis. Testis size, which is an indicator of degree of perturbance during spermatogenesis and used as a proxy for sterility was measured in the four species and their hybrids. It was of same size in D. bipectinata , D. parabipectinata and D. malerkotliana but larger in D. pseudoananassae . In the hybrids involving D. pseudoananassae , testes were atrophied while in other hybrids it was larger. Since, the size of testis does not exhibit uniform pattern of variation in hybrids, it cannot be used as a good indicator for sterility. Therefore, we also measured the size of seminal vesicles (storing organ of sperm) in the four species and their hybrids. Interestingly, the size of seminal vesicles was reduced uniformly in all the hybrids indicating its use as better proxy for sterility. Further, the seminal vesicle size in D. pseudoananassae was smaller than that in the other three species. These observations provide evidence for phylogenetic proximity of D. bipectinata , D. parabipectinata and D. malerkotliana and their remote relationships with D. pseudoananassae .     

Male sterility of the mouse t-complex is due to homozygosity of the distorter genes

Evidence is presented that the male sterility produced by the mouse t-complex is due to interaction of at least three sterility factors. These factors are carried in the same partial haplotypes as the three distorter genes, Tcd-1, Tcd-2, and Tcd-3 and are suggested to be identical with them. When heterozygous, the distorter/sterility genes act on the wild-type form of the responder gene, rendering sperm carrying it nonfunctional, thus leading to high transmission of the t form of the responder. When homozygous, the harmful effects of the distorter genes are stronger and affect both forms of the responder, leading to sterility. If homozygous sterility is an inescapable part of ratio distortion, then the t-lethals confer a selective advantage in removing sterile males from the population. Thus, the relationship between the various properties of the t-complex can now be understood.     

The correction of dysbiotic disorders of the vaginal microflora by using a preparation made from highly adhesive lactobacteria

The effectiveness of a new bacterial preparation obtained from highly adhesive lactobacteria and intended for the correction of dysbiotic disturbances of vaginal microflora was studied in the treatment of 60 pregnant women with dysbacteriosis of the maternal passages. 30 pregnant women were simultaneously treated by the vaginal application of Lactobacterin. The study showed that the use of the preparation of highly adhesive lactobacteria caused the pronounced and stable correction of the microflora of the maternal passages. This correction was manifested by the domination of lactic acid bacterial flora and a decrease in the number of opportunistic microorganisms.