Clipboard, Search History, and several other advanced features are temporarily unavailable. Part 1: background to spermatogenesis, spermatogonia, and spermatocytes. Generating an ePub file may take a long time, please be patient. Biol Reprod. Lipid droplets are cytoplasmic organelles that store neutral lipids and are critically important for energy metabolism. Methods: Canine and Feline Epididymal Semen-A Plentiful Source of Gametes. First detected by Santiago Ramn y Cajal, there are strange droplets in the cytoplasm of cells that are not well understood. The confusion in CD morphology and location between mouse and human spermatozoa may result from the fact that mouse epididymal spermatozoa are usually used for studies, whereas human ejaculated, rather than epididymal, spermatozoa are analyzed. Accessibility We also examined CDs on spermatozoa of three knockout mouse lines with late spermiogenic defects. The cytoplasm also contains Golgi apparatus, and only a small amount of ribosomes and endoplasmic reticulum. Zheng H, Stratton CJ, Morozumi K, Jin J, Yanagimachi R, et al. An aliquot of suspension (5l) was transferred to a chamber slide of 20m depth (Conception Technologies) for observation and movie recording. Introduction. 2021 Oct 14;11(10):2961. doi: 10.3390/ani11102961. An official website of the United States government. 2013 Feb 26;55(1):17. doi: 10.1186/1751-0147-55-17. Supplementary Information accompanies the paper on Asian journal of Andrology website (http://www.nature.com/aja). Their function in energy storage is firmly established and increasingly well characterized. Oko R, Hermo L, Chan PT, Fazel A, Bergeron JJ. METTL21A, a Non-Histone Methyltransferase, Is Dispensable for Spermatogenesis and Male Fertility in Mice. The https:// ensures that you are connecting to the CD, cytoplasmic droplet; WT, wild-type. Motility and percentage of CD-bearing spermatozoa in WT, Ejaculated spermatozoa of C57Bl/6J male mice collected from the female (CD1) reproductive tract immediately after females were mated. While CDs of 61%74% of wild type spermatozoa were located on the flagella (any flagellar domain), only 19%34% of Spem1-null spermatozoa possessed CDs on the flagella (Figure 5b). Cooper TG. This phenomenon may be universal and further examination of CDs in other KO mouse lines with late spermiogenic disruptions may strengthen this claim. Loss-of-function 25.15C), creating clefts or separation artifacts that assist in dif-PTCH mutations are common, and about one-third of these ferentiating basal cell carcinomas from certain appendage tumors mutations consist of CT transitions that are hallmarks of that are also characterized by proliferation of basaloid cells, such UV damage. WY conceived and designed the study. Future proteomic and functional analyses may shed light on the true physiological role of this mysterious organelle. Pea AI, Barrio M, Becerra JJ, Quintela LA, Herradn PG. (a) Total (initial+progressive) motility of spermatozoa collected from the caput, corpus and cauda epididymidis of WT, Tnp1/, Tnp1/Hils1/ and Spem1/ mice. Allow efficient transportation of nutrients across cells in plants and fungi 4. CD, cytoplasmic droplet. CM and CLD metabolism are closely connected processes that are under coordinated molecular regulation [7]. Whereas when obtained from the caput epididymidis the majority of spermatozoa (91%) possessed CDs, the number of CD-bearing spermatozoa decreased to 50% when from the corpus epididymidis, and to only 16% when from the cauda epididymidis (Figure 4b), suggesting that monkey spermatozoa gradually lose their CDs during their epididymal transition from the caput to cauda. (B) DIC microscopic images showing various positions of CDs on monkey epididymal spermatozoa. An investigation of excess residual cytoplasm in human spermatozoa and its distinction from the cytoplasmic droplet. Pie charts showing proportions of the CD locations on spermatozoa collected from the caput, corpus and cauda epididymidis of mice (C57BL/6J, ICR and FVB strains) and monkeys. However, we have noticed that epididymal spermatozoa that display initial (flagellation in situ) and progressive motility mostly possess CDs, whereas spermatozoa without CDs are rarely motile, suggesting that CDs have a role in motility development during sperm epididymal maturation. Yeung CH, Callies C, Rojek A, Nielsen S, Cooper TG. In the present study, we systematically studied the correlation between the presence or absence of CDs and sperm motility in three commonly used laboratory mouse strains and cynomolgus monkeys. In mice, about 30%40% of cauda epididymidal spermatozoa collected into human tubal fluid (HTF) medium never become motile.19 Similarly, about 20%40% of human spermatozoa in ejaculates are immotile,20 suggesting that a large proportion of spermatozoa produced are defective, at least in their ability to develop motility. The .gov means its official. Soler C, Cooper TG, Valverde A, Yniz JL. Most clinicians consider their presence on sperm to be indicative of abnormal sperm, whereas basic scientists consider them to be attributes of normal sperm. We have noticed that spermatozoa produced by KO mice with defective spermiogenesis tend to display aberrant motility characterized by reduced motility that quickly diminishes within a short period of time in vitro.21,22,23 Interestingly, we found that epididymal spermatozoa from these mutant male mice either do not have CDs or their CDs are located in abnormal positions (i.e., are ectopic). Motility and percentage of CD-bearing spermatozoa in WT, Ejaculated spermatozoa of C57Bl/6J male mice collected from the female (CD1) reproductive tract immediately after females were mated. (a) Total (initial+progressive) motility of spermatozoa collected from the caput, corpus and cauda epididymidis of monkeys. The animal protocols were approved by the Institutional Animal Care and Use Committee of the University of Nevada (Reno, NV, USA). However, the total motility of Spem1-null spermatozoa lasted for 30min in HTF medium at 37C, and within 1h all spermatozoa become immotile (data not shown). PMC Clipboard, Search History, and several other advanced features are temporarily unavailable. Abnormal CD formation, e.g., a complete lack of CDs or ectopic CDs, is indicative of defective spermiogenesis. Our data suggest that the CD is a normal organelle transiently present exclusively on epididymal spermatozoa, and normal CD morphology and location are associated with normal motility development during epididymal maturation of spermatozoa. Tnp-1 and Hils1 double KO (Tnp1/; Hils1/) spermatozoa collected from the three regions of the epididymis all displayed significantly reduced initial motility compared with WT spermatozoa (Figure 5a). PLoS One. Alkaline phosphatase in the cytoplasmic droplet of mammalian spermatozoa. Ejaculated spermatozoa of C57Bl/6J male mice collected from the female (CD1) reproductive tract immediately after females were mated. Accessibility However, such mechanism remains unclear in dogs. Isolation and characterization of the cytoplasmic droplet in the rat. The term cytoplasmic droplets is used to mean two different structures by basic scientists and clinicians. As cytoplasmic P -Akt could still be associated with intracellular membranes or the droplet surface, it may be well positioned to modulate this transport process. The term cytoplasmic droplets is used to mean two different structures by basic scientists and clinicians. Retained cytoplasmic droplets (CD) provide the most abundant sperm abnormality in boar and reduce fertility. Spem1-null cauda epididymidal spermatozoa display initial motility and weak progressive motility, neither of which is long lasting (<1h). CDs are believed to play a role in sperm volume adaptation. A mutation study of sperm head shape and motility in the mouse: lessons for the clinic. Pure crowding effects typically are modeled as hard-sphere repulsive interactions that sterically exclude . The recognition that the cell cytoplasm is a highly crowded medium has led to much study and theorizing about the effects of macromolecular crowding on cellular biochemistry (for reviews see Dix and Verkman, 2008 ; Zhou et al., 2008 ). Fabrizio JJ, Rollins J, Bazinet CW, Wegener S, Koziy I, Daniel R, Lombardo V, Pryce D, Bharrat K, Innabi E, Villanobos M, Mendoza G, Ferrara E, Rodway S, Vicioso M, Siracusa V, Dailey E, Pronovost J, Innabi S, Patel V, DeSouza N, Quaranto D, Niknejad A. Biol Open. 2007 Oct;42(5):471-8. doi: 10.1111/j.1439-0531.2006.00809.x. Motility and percentage of CD-bearing spermatozoa in WT, Tnp1/, Tnp1/Hils1/ and Spem1/ mice. Presence of CDs is correlated with sperm motility. Bookshelf Though the distinction between cytoplasmic droplets and excess residual cytoplasm has been made, some studies Samples collected from the corpus epididymis showed higher motility and mitochondrial activity in comparison to the caput sperm. Nucleoprotein transitions during spermiogenesis in mice with transition nuclear protein Tnp1 and Tnp2 mutations. Bethesda, MD 20894, Web Policies FOIA and datasets marked with different letters are statistically significant (P<0.05, n=4). All authors declared no competing financial interests. MeSH Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs-Relationship with Sperm Morphology. The ePub format uses eBook readers, which have several "ease of reading" features TMAs can be analysed and detected on imported files. The presence of cytoplasmic droplets on human sperm was examined using conventional air-dried, fixed and stained sperm smears and in living and fixed wet preparations. Results derived through Excel VBA. Pie charts showing proportions of the CD locations on spermatozoa collected from the caput, corpus and cauda epididymidis of mice (C57BL/6J, ICR and FVB strains) and monkeys. Oxidative Stress in the Male Germline: A Review of Novel Strategies to Reduce 4-Hydroxynonenal Production. (. (b) Percentage of CD-bearing cells of the total motile spermatozoa collected from the caput, corpus and cauda epididymidis of monkeys. Moving to the beat: a review of mammalian sperm motility regulation. ( A ) DIC, CD positional changes during epididymal transit in mice and monkeys. Arrows point to CDs, which can be located at the neck (a), the middle piece (b), the mid-principal piece junction (c), and the principal/end piece (d) (scale bar=5m). They respond to stimuli initiated either on the cell surface or in the cytoplasm as conditions change. 2022 Feb 9;23(4):1942. doi: 10.3390/ijms23041942. Biol Psychiatry. PMC Disclaimer, National Library of Medicine Although the cytoplasm of spermatids is removed at the end of spermiogenesis, a tiny portion is usually retained in the sperm flagellum, which is termed the cytoplasmic droplet (CD) in mammals. Reprod Domest Anim. This site needs JavaScript to work properly. Cooper TG, Yeung CH, Fetic S, Sobhani A, Nieschlag E. Cytoplasmic droplets are normal structures of human sperm but are not well preserved by routine procedures for assessing sperm morphology. Nat Rev Urol. Arrdc4-dependent extracellular vesicle biogenesis is required for sperm maturation. Loss-of-function mutations in seipin account for a severe form of congenital lipodystrophy (Berardinelli-Seip congenital lipodystrophy) and . Ahmed O, Lehloenya K, Mphaphathi M, Hassen A. (b) Percentage of spermatozoa bearing CDs on any region of the flagellum, collected from the caput, corpus and cauda epididymidis of WT, Tnp1/, Tnp1/Hils1/ and Spem1/ mice. official website and that any information you provide is encrypted Interestingly, the majority of monkey caput epididymidal spermatozoa (92%) displayed CDs on the neck, whereas the majority of CDs (88%89%) of corpus and cauda epididymidal spermatozoa were found on the mid-piece (Figure 2), demonstrating a descending movement of CDs as spermatozoa transited through the epididymis in monkeys. J Histochem Cytochem. Cyto means cell and plasm means stuff, so together it's cell staff. In contrast, cauda epididymidal spermatozoa displayed mostly progressive (i.e., forward) motility after incubation in the HTF medium (Supplementary Movie 3). 2006 Feb;41(1):63-7. doi: 10.1111/j.1439-0531.2006.00621.x. Accessibility 2012 Nov 17;10:92. doi: 10.1186/1477-7827-10-92. The Rev protein mediates cytoplasmic transport of viral mRNAs that code for HIV-1 structural protein; It is, therefore, not surprising that spermatozoa from both KO lines display drastically reduced total motility. The Gregarious Lipid Droplet*. MeSH (a) Percentage of CD-bearing motile and immotile spermatozoa from the caput epididymidis of C57BL/6J, ICR and FVB mouse strains. It is proposed that a change in terminology is required: 'cytoplasmic droplet' should be used to describe normal droplets associated with functional sperm produced by normal spermatogenesis and 'excess residual cy toplasm' is suggested to describe abnormal droplet associated with dysfunctional sperm that are products of faulty sperMatogenesis. Epub 2014 Dec 16. A total of five adult KO mice with each of the four genotypes (wild-type, Tnp1/, Tnp1/Hils1/ and Spem1/) were used to collect data on the presence or absence of CDs and ectopic CDs on epididymal spermatozoa. Nord Vet Med. Acquisition of volume regulatory response of sperm upon maturation in the epididymis and the role of the cytoplasmic droplet. Online ahead of print. 2021 Aug 31;11(9):2570. doi: 10.3390/ani11092570. Data are presented as means.e.m. Anti-AUP1 Antibody (PACO07954) from Assay Genie is a high quality polyclonal antibody developed for research and validated in WB, IHC applications PMC There has been considerable interest in them lately, as they seem to be important to the activities and functions of the cell. CD, cytoplasmic droplet. The .gov means its official. CD positional changes during epididymal transit in mice and monkeys. Comparative study of boar sperm coming from the caput, corpus, and cauda regions of the epididymis. In summary, the data presented here suggest that the CD is a normal organelle exclusively present on epididymal spermatozoa in mice and monkeys, and normal CD morphology, location and migration patterns are correlated with motility development potential during epididymal sperm maturation. Llavanera M, Delgado-Bermdez A, Mateo-Otero Y, Padilla L, Romeu X, Roca J, Barranco I, Yeste M. Antioxidants (Basel). Spem1-null spermatozoa display head-bent-back' phenotype with 100% penetrance.21 Electron microscopic analyses suggest that cytoplasmic removal may have been delayed in the absence of SPEM1 in elongated spermatids, leading to CD-like structures enveloping the bent head and neck in Spem1-null spermatozoa.21,29,30 The existence of CDs was further confirmed by immunofluorescence staining of proteins known to be highly abundant in CDs, including SPEM1, ubiquitin and 15-lipoxygenase.21 Three KO mouse lines (Tnp1/, Tnp1/Hisl1/ and Spem1/) were housed in a temperature- and humidity-controlled animal facility in the University of Nevada, Reno, NV, USA, with free access to water and food. Would you like email updates of new search results? We have generated several lines of KO mice with late spermiogenic defects with no histologically discernable disruptions of the seminiferous epithelium.23 We have long noticed that epididymal spermatozoa in those KO male mice either do not bear CDs or have CDs that are not located in the common positions (i.e., middle piece or mid-principal piece junction) on the flagella.21,22,25,26 We analyzed three such KO mouse lines in this study, and our results support the notion that a lack of CDs or ectopic CDs appears to be indicative of spermiogenic defects in the absence of seminiferous epithelial disruption at histological level. Instead they displayed initial motility characterized by non-progressive flagellation in situ in the HTF medium (Supplementary Movies 1 and 2). Positions of CDs on mouse and monkey epididymal spermatozoa. Infertility in a dog due to proximal cytoplasmic droplets in the ejaculate: investigation of the significance for sperm functionality in vitro. More interestingly, the presence of CDs is correlated with the potential of motility development. Cooper TG, Yeung CH, Fetic S, Sobhani A, Nieschlag E. Hum Reprod. MeSH Evaluation of the Morphometry of Sperm from the Epididymides of Dogs Using Different Staining Methods. Bookshelf Rengan AK, Agarwal A, van der Linde M, du Plessis SS. Movie clips of different fields were recorded by using an inverted phase contrast microscope equipped with a video camera (AxioVision; Carl Zeiss, Oberkochen, Germany). These results suggest that defective spermiogenesis, although it may not be histologically discernable, often produces spermatozoa without CDs or with ectopic CDs. 2019 Jan 15;85(2):164-171. doi: 10.1016/j.biopsych.2018.09.007. The protocol for euthanizing monkeys was approved by the Institutional Animal Care and Use Committee which assures compliance with the United States Department of Agriculture, Public Health Service Office of Laboratory Animal Welfare Policy and the Animal Welfare Act (Charles River Laboratories, Preclinical Services, Sparks, NV, USA). Although CDs could be found in any position along the flagella of epididymal spermatozoa in mice (Figure 1A), the majority (80%97%) of CDs were found on the middle piece of sperm flagella in all three strains of mice analyzed (Figure 2). 300mosmol l1 in the uterine cavity vs. 440mosmol l1 in the cauda epididymidis).10. Noteworthy, recruitment of FUS into phase separated RNP granules, e.g . Hermo L, Pelletier RM, Cyr DG, Smith CE. Hermo L, Dworkin J, Oko R. Role of epithelial clear cells of the rat epididymis in the disposal of the contents of cytoplasmic droplets detached from spermatozoa. 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