Normal sperm head morphometric reference values in fertile Asian males

Recombinant inbred strains were developed from reciprocal crosses between two inbred strains of mice (CBA and KE) differing in sperm head shape, proportion of normal sperm heads (CBA, 95%; KE, 78%) and fertilization efficiency (CBA, 100% of fertilized ova; KE, 72%), to determine whether the indices of sperm morphology and function were correlated. The following parameters were analysed in recombinant inbred and progenitor strains: index of sperm head shape (head width in the middle of its length/head length), percentage of abnormal sperm heads, percentage of spermatozoa with progressive movements, efficiency of penetration of hyaluronic acid polymer (Sperm Select) and percentage of fertilized ova after mating males from the tested strains with females from an outbred stock. For each investigated character, recombinant inbred strains, recombinant inbred EXCB and CBXE, could be divided into at least three categories: KE-like, CBA-like and intermediate, suggesting that in each case a minimum of two genes was involved. Recombinant strains derived from the reciprocal crosses of progenitor strains differed only with respect to the proportion of abnormal sperm heads, showing the involvement of the Y chromosome in determining this character. Penetration into Sperm Select was significantly correlated both with fertilization efficiency and sperm motility, while correlation with the proportion of normal spermatozoa did not reach the level of significance. However, there was a significant negative correlation of both sperm abnormalities and the incidence of supplementary spermatozoa in the perivitelline space with the index of sperm head shape.(ABSTRACT TRUNCATED AT 250 WORDS)

Sperm exhibit extraordinary levels of morphological diversification across the animal kingdom. In songbirds, sperm have a helically shaped head incorporating a distinct acrosomal membrane or "helical keel," the form and extent of which varies across species. The functional significance of this helical shape, however, remains unknown. Using scanning electron microscopy, we quantified inter- and intraspecific variation in sperm head morphology across 36 songbird species (Passeriformes: Passerida). Using phylogenetic comparative methods, we investigated the relationship between sperm head morphology and both sperm swimming speed and the frequency of extra-pair young (EPY). We found that species whose sperm had a relatively more pronounced helical form (i.e., long acrosome, short nucleus, wide helical membrane, and a more pronounced waveform along the sperm head "core") had faster-swimming sperm. We found no evidence of a relationship between interspecific variation in sperm head morphology and EPY, although we did find that among- and within-male variation in sperm head traits were negatively correlated with EPY. Applying principles of fluid mechanics, we discuss how the helical form of the sperm head may influence swimming speed, and suggest that further studies considering aspects of sperm morphology beyond sperm length are needed to improve our understanding of sperm structure-function relationships.

The main objective of this study was to evaluate the potential genetic effects of SEPT14 on male infertility through sequencing the SEPT14 coding region. To address this research gap, 254 men with sperm abnormalities and 116 normozoospermic men were recruited, and the whole-coding regions of SEPT14 were sequenced. Two heterozygous mutations, p.Ala123Thr (3/254 vs. 0/116) and p.Ile333Thr (3/254 vs. 0/116), were identified in these cases. A high percentage of defective sperm heads was found in sperm with mutated SEPT14. Both mutations are highly evolutionarily conserved among vertebrates. The results of a fine morphological and chromatin structural analysis indicated severely malformed sperm heads with abnormal chromatin packaging through transmission electron microscopy and Toluidine blue staining. Compared with controls, high DNA fragmentation was demonstrated in sperm from cases carrying SEPT14 mutations using the comet assay. In addition, these two mutations in SEPT14 affected its polymerization ability in vitro. These data revels that the two SEPT14 missense mutations impaired sperm head morphology and induced DNA damage. Our study suggests that genetic variant of SEPT14 is one of the effects for human sperm formation and male fertility.

Inverse correlation between chromatin condensation and sperm head size in a case of enlarged sperm heads

Introduction Studies of the association between polychlorinated biphenyls (PCBs) and sperm morphology are contradictory. Two possible reasons are 1) different PCB congeners and congener mixtures may have different effects and 2) the standard measure of percent abnormal sperm combines several outcomes, e.g. abnormal heads and tails. Here we explore the effect of prenatal exposure to PCBs on 3 measures of abnormal sperm morphology. Methods In a follow-up of 196 men from the Child Health and Development Studies cohort, we evaluated associations between prenatal exposure to total PCBs and 15 PCB congeners, both individually and grouped according to chlorination patterns and proposed biological mechanism, and percent abnormal sperm (total, head and tail) at mean age 43. PCBs were measured in stored maternal blood taken immediately postpartum. Sperm morphology was measured in semen collected from adult male offspring and analyzed according to WHO criteria. All analyses adjusted for maternal total lipids, race, income and coffee consumption, and offspring age, smoking, and perceived stress. Results Each ng/mL increase in total PCBs in maternal blood was associated with an increase of 0.92 percent total normal sperm (95% CI: 2.35, -0.51), driven by a reduction in head abnormalities (bhead= -1.30 [-2.31, -0.29] but with contrasting evidence for tail abnormalities (btail=0.49 [-0.38, 1.36]). Among total, grouped and individual PCBs, prenatal exposure was generally associated with fewer head abnormalities and more tail abnormalities and the estimated association was strongest for di ortho substituted PCBs Conclusions Prenatal PCB exposure may have opposite effects on sperm head and tail morphology, warranting further research into the influence of PCBs on the development of fetal Sertoli cells. Prenatal PCB exposure may create an unfavorable environment for production or survival of sperm with head abnormalties. The clinical consequences of this finding are uncertain.

Testicular structure of 9 species and sperm head morphology of 19 species of Cottidae were observed in order to clarify relationships between morphological characteristics of the male reproductive organ and reproductive mode (copulation or non-copulation). Morphological structure of the testis was divided into the following five types based on the sperm transfer and reservoir system: (1) a non-duct type in which the sperm duct is not a distinct exterior structure, but the tube for sperm transport traverses along the testis as an interior structure; (2) an anterior duct type with distinct anterior sperm ducts traversing along the testis; (3) a posterior duct type with distinct anterior sperm ducts traversing along the dorsal hilus of testis and posterior sperm ducts extending to the rear of the testis; (4) an anterior duct posterior vesicle type with distinct anterior sperm ducts traversing along the testis, and the right and left sperm ducts fusing in the rear of testis, forming the seminal vesicle; (5) a non-duct posterior vesicle type in which sperm ducts do not accompany the testis, and the testis and seminal vesicle are connected directly or through posterior sperm ducts. It is thought that in Cottidae the non-duct type of reproductive organ is primitive, and the anterior duct type is common to all non-copulating species. The testes and accompanying seminal vesicle were seen only in copulating species. Sperm head morphology was divided into three types according to the length/width ratio: oval type ≤2, intermediate type >2 and ≤3, and slender type >3. The type of sperm head corresponded closely to the reproductive mode; non-copulating species had oval sperm head, and copulating species had intermediate or slender ones. These results suggest that the structure of the testis and the morphology of the sperm head evolved from testes with anterior sperm ducts and oval sperm heads to testes with an associated seminal vesicle and slender sperm heads in association with the evolution from non-copulatory to copulatory reproduction in Cottidae.

Mammalian sex chromosomes are enriched for large, nearly-identical, palindromic sequences harboring genes expressed predominately in testicular germ cells. Discerning if individual palindrome-associated gene families are essential for male reproduction is difficult due to challenges in disrupting all copies of a gene family. Here we generate precise, independent, deletions to assess the reproductive roles of two X-linked palindromic gene families with spermatid-predominant expression, 4930567H17Rik and Mageb5. Sequence analyses reveals mouse 4930567H17Rik and Mageb5 are orthologs of human HSFX3 and MAGEB5, respectively, where 4930567H17Rik/HSFX3 is harbored in a palindrome in humans and mice, while Mageb5 is not. Additional sequence analyses show 4930567H17Rik and HSFX3 are rapidly diverging in rodents and primates, respectively. Mice lacking either 4930567H17Rik or Mageb5 gene families do not have detectable defects in male fertility, fecundity, spermatogenesis, or in gene regulation, but do show differences in sperm head morphology, suggesting a potential role in sperm function. We conclude that while all palindrome-associated gene families are not essential for male fertility, large palindromes influence the evolution of their associated gene families.

Detailed descriptions of mammalian sperm morphology across a range of closely related taxa are rare. Most contributions have been generalized descriptions of a few distantly related mammalian species. These studies have emphasized a generalized ungulate sperm morphology, but have not underscored several important morphological differences in ungulate sperm, such as head shape. The present study is the first to document descriptions of sperm head morphology using cold field-emission scanning electron microscopy (FE-SEM) for a large number of closely related mammalian species. In total, the sperm of 36 species in three orders: Artiodactyla (even-toed ungulates), Cetacea (whales, porpoises, and dolphins), and Perissodactyla (odd-toed ungulates) were examined to gather new information relevant to the debate about the phylogenetic placement of cetaceans relative to terrestrial ungulates. In all species examined, the sperm heads were generally flattened and ovate in shape with a distinct apical ridge, although considerable variation in sperm head shape was detected, both within and between orders. In artiodactylans, the sperm head was uniformly flat in lateral view, whereas perissodactylan and cetacean sperm heads showed a distinct posterior thickening. In both artiodactylans and perissodactylans, the mitochondria were elongate and wound in a tight helix around the midpiece, whereas in cetaceans the mitochondria were rounded and appeared to be randomly arranged around the midpiece. Additionally, prominent ridges running along the anterior-posterior axis were observed in the postacrosomal region of the sperm head in four species of cetaceans. These ridges were not observed in any of the terrestrial ungulates examined. Pits or fenestrations were detected in the postacrosomal region in most artiodactylan species examined; these structures were not detected in perissodactylans or cetaceans. The equatorial segment of the acrosome was detected in the artiodactylan species examined, tentatively identified in perissodactylans, but not found in cetaceans. Its shape and location are described for relevant taxa. The presence of a recently reported substructure within the equatorial segment (the equatorial subsegment; Ellis et al. [2002] J Struct Biol 138:187-198) was detected in artiodactylans, and its shape is described for the species examined.

In vitro, He-Ne laser show a modifying response of cells to ionizing radiations. So there is a need to investigate this property in vivo.To investigate the effect of He-Ne laser (632.8nm) pre and post irradiation against the UVC (253.7nm) irradiation in vivo on the sperm morphology, and DNA of testis in Mouse. In this study Twenty adult albino male mice were divided into four groups, the first group was He-Ne laser irradiated, and the second group was UVC irradiated. The third group was pre-irradiated by He-Ne laser followed by UVC irradiation, with one hour time interval between the two irradiations. The fourth group was pre-irradiated by UVC then by He-Ne laser, with one hour separating the two irradiations. The He-Ne laser per-irradiation show a protection properties, which appeared on both sperm head abnormality and DNA damage against UVC light irradiation. Four sperm head abnormality features were found after each type of irradiation. But the He-Ne laser preirradiation against UVC irradiation lower the frequencies of sperm head abnormality, farther more reduce the DNA testis damaging. Both UVC and He-Ne laser irradiations revealed a toxic effect on sperm head morphology, but the HeNe laser was found less effective. The He-Ne laser protective properties was observed only when He-Ne laser irradiation was applied before the UVC irradiation.

This chapter summarizes the scientific and clinical utility of the nuclear and cytoplasmic sperm biomarkers in the diagnosis and treatment of male infertility. These biomarkers were developed and used by the Yale Sperm Physiology Laboratory in the past 15 years. These biomarkers tested and were proven to reflect the degree of sperm development and fertilizing potential, including cytoplasmic retention measured by sperm creatine kinase content, chromatin structure, the key factor of HSPA2 chaperon protein concentration, DNA chain integrity, chromosomal aneuploidies, sperm head and tail shape and length, and most important, spermiogenetic remodeling of the plasma membrane that is the basis of the sperm-hyaluronic acid-binding assay in the Andrology Laboratory, and of the hyaluronic acid-mediated ICSI sperm selection in the Embryology laboratory. Some aspects of ICSI sperm selection by sperm shape combined with high magnification are also discussed.

DNA fragmentation and sperm head morphometry in cat epididymal spermatozoa

The morphology of the spermatozoon of representative species of the subfamily Nesomyinae (Muroidea: Nesomyidae), a monophyletic group of rodents endemic to Madagascar, was examined by light and electron microscopy to determine the sperm head shape and tail length across the species. Marked interspecific differences were found to occur in both the form of the sperm head and length of the tail. The species that possess a sperm head with an apical hook, which largely contains acrosomal material, generally displayed longer sperm tails, and a species with a spatulate sperm head had the shortest tail. The association between sperm head shape and tail length mirrors that previously found in Eurasian and Australasian murine rodents. Thus, the repeated association between sperm head shape and tail length across these groups of muroid rodents clearly indicates a functional relationship between these two features. A comparison of sperm morphology of the nesomyines to that of related muroid rodents on the mainland of Africa suggests that the possession of an apical hook is the ancestral condition.

Sperm display remarkable morphological diversity among even closely related species, a pattern that is widely attributed to postcopulatory sexual selection. Surprisingly few studies have used phylogenetic analyses to discern the details of evolutionary diversification in ornaments and armaments subject to sexual selection, and the origins of novel sperm traits and their subsequent modification are particularly poorly understood. Here we investigate sperm evolution in diving beetles (Dytiscidae), revealing dramatic diversification in flagellum length, head shape, presence of sperm heteromorphism, and the presence/type of sperm conjugation, an unusual trait where two or more sperm unite for motility or transport. Sperm conjugation was found to be the ancestral condition in diving beetles, with subsequent diversification into three forms, each exhibiting varying degrees of evolutionary loss, convergence, and recurrence. Sperm head shape, but not length or heteromorphism, was found to evolve in a significantly correlated manner with conjugation, consistent with the different mechanisms of head alignment and binding required for the different forms of conjugation. Our study reveals that sperm morphological evolution is channeled along particular evolutionary pathways (i.e., conjugate form), yet subject to considerable diversification within those pathways through modification in sperm length, head shape, and heteromorphism.

Mammalian spermatozoa, particularly those of rodent species, are extremely complex cells and differ greatly in form and dimensions. Thus, characterization of sperm size and, particularly, sperm shape represents a major challenge. No consensus exists on a method to objectively assess size and shape of spermatozoa. In this study we apply the principles of geometric morphometrics to analyze rodent sperm head morphology and compare them with two traditional morphometry methods, that is, measurements of linear dimensions and dimensions-derived parameters calculated using formulae employed in sperm morphometry assessments. Our results show that geometric morphometrics clearly identifies shape differences among rodent spermatozoa. It is also capable of discriminating between size and shape and to analyze these two variables separately. Thus, it provides an accurate method to assess sperm head shape. Furthermore, it can identify which sperm morphology traits differ between species, such as the protrusion or retraction of the base of the head, the orientation and relative position of the site of flagellum insertion, the degree of curvature of the hook, and other distinct anatomical features and appendices. We envisage that the use of geometric morphometrics may have a major impact on future studies focused on the characterization of sperm head formation, diversity of sperm head shape among species (and underlying evolutionary forces), the effects of reprotoxicants on changes in cell shape, and phenotyping of genetically-modified individuals.

Sperm heads contain not only the nucleus but also the acrosome which is a distinctive cap-like structure located anterior to the nucleus and is derived from the Golgi apparatus. The Golgi Associated RAB2 Interactors (GARINs; also known as FAM71) protein family shows predominant expression in the testis and all possess a RAB2-binding domain which confers binding affinity to RAB2, a small GTPase that is responsible for membrane transport and vesicle trafficking. Our previous study showed that GARIN1A and GARIN1B are important for acrosome biogenesis and that GARIN1B is indispensable for male fertility in mice. Here, we generated KO mice of other Garins, namely Garin2, Garin3, Garin4, Garin5a, and Garin5b (Garin2-5b). Using computer-assisted morphological analysis, we found that the loss of each Garin2-5b resulted in aberrant sperm head morphogenesis. While the fertilities of Garin2-/- and Garin4-/- males are normal, Garin5a-/- and Garin5b-/- males are subfertile, and Garin3-/- males are infertile. Further analysis revealed that Garin3-/- males exhibited abnormal acrosomal morphology, but not as severely as Garin1b-/- males; instead, the amounts of membrane proteins, particularly ADAM family proteins, decreased in Garin3 KO spermatozoa. Moreover, only Garin4 KO mice exhibit vacuoles in the sperm head. These results indicate that GARINs assure correct head morphogenesis and some members of the GARIN family function distinctively in male fertility.