Molecular Reproduction and DevelopmentVolume 88, Issue 10 p. 647-649 CORRESPONDENCE Male contraception: Are we seeing the end of the beginning? Harvey M. Florman, Corresponding Author Harvey M. Florman harvey.florman@umassmed.edu orcid.org/0000-0002-2399-3493 Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester, Massachusetts, USA Correspondence Harvey M. Florman, Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester, MA 01655, USA. Email: harvey.florman@umassmed.eduSearch for more papers by this author Harvey M. Florman, Corresponding Author Harvey M. Florman harvey.florman@umassmed.edu orcid.org/0000-0002-2399-3493 Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester, Massachusetts, USA Correspondence Harvey M. Florman, Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester, MA 01655, USA. Email: harvey.florman@umassmed.eduSearch for more papers by this author First published: 30 August 2021 https://doi.org/10.1002/mrd.23531Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume88, Issue10October 2021Pages 647-649 RelatedInformation

January 1919 was a busy month. Hostilities of World War I ended in November 1918 but a state of war persisted. On January 8, 1919, Woodrow Wilson enumerated the postwar aims of the United States in his 14 points and this was followed later in the month by the opening of the Paris Peace Conference. Despite this, widespread chaos continued. Consider Europe, there were bloody street battles in Berlin and elsewhere in Germany from January 5 to 12, 1919 between Spartacist militias on one side and Freikorps and government units on the other. Attempted coups were mounted in the Balkans and Poland. In Ireland, the First Dail issued a “Message to the Free Nations of the World” on January 21, 1919, in which independence was declared from Great Britian, and fighting began the same day with the Soloheadbeg Ambush. Glasgow saw the Battle of George Square between striking labor groups on one side against police and military units on January 31, 1919. Ongoing conflicts included the Baltic wars of independence from Russia, as well as the Russian civil war between White and Red forces. And It was not only Europe: there was, for example, the Semana Tragica (January 7–14, 1919) in Argentina, with fighting between Anarchist and Communist forces on one side against federalist police. For cell and developmental biologists that January of 1919 was also notable for the monthly issue of The Biological Bulletin. It included three articles by Ernest Everett Just which considered various aspects of fertilization in Echinarachnius parma, the common sand dollar. The first of those reported details of the egg's cortical response to insemination, including a dissection of the functional consequences of this reaction (Just, 1919). That paper marks a conceptual turning point in our still-emerging understanding of how the egg limits sperm entry. Why this was the case requires some background. Let us look further back another 40 years. The last several decades of the 19th century were a critical period for cell and developmental biology. New methods of fixation and staining of tissues were developed and disseminated; some of these approaches were particularly well known, such as the Golgi “black reaction” (Peters, 2007), but in fact, a large number of techniques first became available to the general community (Lee, 1885). In addition, advances in optics and glass chemistry—notably from Jena (then in the Duchy of Saxe-Weimar-Eisenach and today in the Federal State of Thuringia in Germany) by the commercial collaboration of Carl Zeiss, Otto Schott, and Ernst Abbe—lead to the development of immersion apochromatic lenses of high numerical aperture and of advanced condensers. The result was a light microscope of then-unparalleled resolution and clarity. These technological developments contributed to an era of conceptual advances in histology and cytology. It was at this time that two students of Ernst Haeckel-Oscar Hertwig and Hermann Fol—are generally credited with independently discovering the fertilization of animal eggs by sperm, although this history may be more complex than is often presented (Briggs & Wessel, 2006). Both observed the fusion of echinoderm pronuclei although Fol went further and actually described sperm entry into eggs (Fol, 1877, 1879; Hertwig, 1877). In addition, he first articulated the difference between physiological and pathological polyspermy; in taxa of the former group normal development occurs in the presence of multiple sperm-derived pronuclei with the egg cytoplasm, while in the latter group multiple sperm pronuclei result in a halting of development. This distinction provided a valuable tool for the analysis of cellular processes: for example, studies with polyspermic eggs provided Boveri with essential data leading to the development of the chromosome theory of inheritance (Baltzer, 1964; Laubichler & Davidson, 2008; Opitz, 2016; Wilson, 1925). Finally, Fol described the elevation of the fertilization envelope in detail and, importantly, concluded that this provided a cellular mechanism for the block to polyspermy (Fol, 1877, 1879). Experimental proof for a link between the expansion of the fertilization envelope and the block to polyspermy was soon provided by the pharmacological experiments of Oscar and Richard Hertwig (Hertwig & Hertwig, 1887). The role of the fertilization envelope in the prevention of polyspermic fertilization was widely (Heilbrunn, 1913; Lillie, 1919; Wilson, 1925), though not universally (Loeb, 1915a, 1915b), accepted. It is here that Just enters the story. His insight, based on careful experimental design and meticulous animal maintenance, was that the elevation of the fertilization envelope was too slow to account for the block to polyspermy (Just, 1919; as noted by a sharp-eyed reviewer of this essay, Just suggests in a footnote on page 7 of that 1919 report that Oskar Hertwig may have anticipated a role for the egg cortex in the block to polyspermy as early as 1878.) From here we trace the distinction between the cell surface and egg coat blocks to polyspermy. This also provided an experimental approach that permitted examination of underlying mechanisms. The cell surface block, which is often termed the “fast block,” was studied extensively in the following decades (Clark, 1936; Gray, 1922; Rothschild, 1954; Rothschild & Swann, 1952; Tyler, 1948), culminating in Jaffe's landmark demonstration of the role of egg membrane potential in the control of egg fertilizability (Jaffe, 1976; Nuccitelli & Grey, 1984). Yet, some taxa, including mammals, have egg surface blocks that are not mediated by membrane potential (Florman & Fissore, 2015; Jaffe, Sharp, & Wolf, 1983), and in those cases, a cellular mechanism is not yet understood. A slower block mediated by the egg coat was also the subject of intense interest in a range of taxa, including mammals (Austin & Braden, 1953a, 1953b), and subsequently shown to be based on biochemical modifications in the egg coat (Bleil, Beall, & Wassarman, 1981; Burkart, Xiong, Baibakov, Jimenez-Movilla, & Dean, 2012; Foerder & Shapiro, 1977; Wong & Wessel, 2006). What follows is a consideration of our understanding of polyspermy regulation today. Just's work provides a rationale for this thematic issue, as it definitively established a role for the egg cortex in the echinoderm block to polyspermy (Just, 1919). Yet this is not intended as a retrospective on Just. Byrnes does conside the role of Just in the history of biology, and readers interested in more details about this remarkable man and scientist are referred to other work by Byrnes (Byrnes, 2009; Byrnes & Newman, 2014), as well as available articles and monographs (Crow, 2008; Manning, 1985). Rather, our focus here is on the consequences of Just's report. Wozniak and Carlson discuss new information on the egg surface ion channels that drive the block to polyspermy, while the egg surface and egg coat blocks are reviewed by Evans and by Fahrenkamp et al., respectively. As discussed previously, Fol had noted that in some taxa polyspermy is compatible with development. This is explored in animals by Iwao et al. and in plants by Tekleyohans and Groβ-Hardt and by Toda and Okamoto. The author declares that there are no conflict of interests.

Mammalian oocytes are enveloped by the zona pellucida (ZP), an extracellular matrix of glycoproteins. In sperm, stimulation with ZP proteins evokes a rapid Ca(2+)influx via the sperm-specific, pH-sensitive Ca(2+)channel CatSper. However, the physiological role and molecular mechanisms underlying ZP-dependent activation of CatSper are unknown. Here, we delineate the sequence of ZP-signaling events in mouse sperm. We show that ZP proteins evoke a rapid intracellular pH(i)increase that rests predominantly on Na+/H(+)exchange by NHA1 and requires cAMP synthesis by the soluble adenylyl cyclase sAC as well as a sufficiently negative membrane potential set by the spem-specific K(+)channel Slo3. The alkaline-activated CatSper channel translates the ZP-induced pH(i)increase into a Ca(2+)response. Our findings reveal the molecular components underlying ZP action on mouse sperm, opening up new avenues for understanding the basic principles of sperm function and, thereby, mammalian fertilization.

Enkurin was identified initially in mouse sperm where it was suggested to act as an intracellular adaptor protein linking membrane calcium influx to intracellular signaling pathways. In order to examine the function of this protein, a targeted mutation was introduced into the mouse Enkurin gene. Males that were homozygous for this mutated allele were subfertile. This was associated with lower rates of sperm transport in the female reproductive tract, including reduced entry into the oviduct and slower migration to the site of fertilization in the distal oviduct, and with poor progressive motility in vitro. Flagella from wild-type animals exhibited symmetrical bending and progressive motility in culture medium, and demembranated flagella exhibited the "curlicue" response to Ca2+ in vitro. In contrast, flagella of mice homozygous for the mutated allele displayed only asymmetric bending, nonprogressive motility, and a loss of Ca2+-responsiveness following demembrantion. We propose that Enkurin is part of a flagellar Ca2+-sensor that regulates bending and that the motility defects following mutation of the locus are the proximate cause of subfertility.

Molecular Reproduction and Development (MRD) turns 40 years old in 2018. It appeared in 1978 under the guise of a different name –Gamete Research– as a vision of Ralph B.L. Gwatkin that “information exchange will be facilitated between those working on gametes and early development in the various phyla”. That mission was reflected in the contents of the first issue: to wit, a review on enzymatic events during mammalian fertilization and reports on fertilization, ovulation, and oocyte maturation in a range of mammalian and non-mammalian species. Although the mission and the captain where the same, the Journal was rechristened as MRD in 1988. For the last decade MRD was guided by Gary M. Wessel. During this period the Journal content continued in the traditional areas of gamete biology and fertilization in a wide range of systems, and expanded to reflect Gary's wide interests in areas such as evolutionary biology. Of note, Gary fostered greater interaction between the Journal and the research community through the sponsorship of meetings. As we go into the next phase, MRD will continue with the mission established by Ralph and refined by Gary. I hope to expand ongoing efforts in several areas, including what might be called “ecological reproduction and development” − that is, the adaptation of gametes and embryos to the external environment or, in the case of taxa that fertilize internally, to the female reproductive tract. We are also particularly interested in the ways that different taxa solved the conundrums of reproduction. Finally, I would like to emphasize the Correspondence as a vehicle for researchers to communicate their data. Firstly, Gary recognized that negative results may sometimes be of value to the community, and the brief style of a Correspondence is perfect for these well-controlled datasets. Secondly, the issues of experimental reproducibility in science constantly plague the field, but confirmatory data are often buried in larger manuscripts or not deemed worthy of publication. The stand-alone Correspondence is a perfect option for such studies. As with all manuscripts, data written for a Correspondence (see our Author Guidelines online) will be evaluated on a case-by-case basis. Let's all thank Ralph and Gary for their outstanding leadership and foresight during the past four decades, and look forward to the next decade of MRD. Harvey M. Florman Editor-in-Chief

Bayard T, Storey (July 13, 1932-June 4, 2017) spent his academic career at the University of Pennsylvania School of Medicine. After training in chemistry (B.S. in Chemistry from Harvard University; M.S. in Chemical Engineering from the Massachusetts Institute of Technology; Ph.D. in Chemistry from Harvard University), he spent several years working on ion-exchange resins at Rohm and Haas in Philadelphia. Only in 1965 did he return to academia as a 33-year-old post-doctoral fellow, with Britton Chance at the Eldridge Reeves Johnson Foundation for Medical Physics at the University of Pennsylvania. Bayard's initial interests in the mitochondrial respiratory chain lead to his recognition that sperm, with mitochondria tethered to the flagellum, provided a unique model. Thus began the parts of his career with which we are most familiar: studies of sperm physiology that encompassed bioenergetics, membrane lipid peroxidation, cryopreservation, and interaction with eggs. He brought the approaches he learned at the Johnson Foundation, where he focused on time resolving mitochondrial electron transfer events, to reproductive physiology. Today we routinely use fluorescent probes to follow events in gametes; however in the 1970s, Bayard, along with Henry Lardy (University of Wisconsin), were among the first to develop real-time assays to study fertilization-related events in living sperm. The reproductive biology community recognized Bayard in many ways, including electing him to organize a Gordon Conference on Fertilization and the Activation of Development and honoring him with the Distinguished Andrologist Award from the American Society of Andrology. Those of us who worked with him, were trained by him, or knew him (there were many) also remember his enthusiasm, wit, charm, and how he generously gave his time to help others. Such assistance could be with a particular experimental issue or a spirited and witty debate of controversies in our field or politics, often around a table with wonderful wines. He was particularly concerned with helping young investigators begin and advance their careers and was a strong advocate for the funding and support of reproductive biology. He is sorely missed, and will forever be remembered. Harvey Florman • University of Massachusetts Medical School, Worcester MA George Gerton • University of Pennsylvania School of Medicine, Philadelphia PA Gregory Kopf • fhi360, Durham NC Pablo Visconti • University of Massachusetts, Amherst MA

Cilia are organelles specialized for movement and signaling. To infer when during evolution signaling pathways became associated with cilia, we characterized the proteomes of cilia from sea urchins, sea anemones, and choanoflagellates. We identified 437 high-confidence ciliary candidate proteins conserved in mammals and discovered that Hedgehog and G-protein-coupled receptor pathways were linked to cilia before the origin of bilateria and transient receptor potential (TRP) channels before the origin of animals. We demonstrated that candidates not previously implicated in ciliary biology localized to cilia and further investigated ENKUR, a TRP channel-interacting protein identified in the cilia of all three organisms. ENKUR localizes to motile cilia and is required for patterning the left-right axis in vertebrates. Moreover, mutation of ENKUR causes situs inversus in humans. Thus, proteomic profiling of cilia from diverse eukaryotes defines a conserved ciliary proteome, reveals ancient connections to signaling, and uncovers a ciliary protein that underlies development and human disease.

The current Editorial Board of Molecular Reproduction and Development would like to express our gratitude to Gary Wessel for his outstanding decade of editorship of the journal. Under Gary's direction, MRD became a “player” in developmental biology and reproductive biology publishing, with excellent research articles and reviews contributing to its increased visibility and impact factor. It is a difficult task to raise the profile of a journal, particularly in these times of proliferating journals and ultra-competitive publishing, but Gary did it! His efforts to recruit exciting and impactful articles, to demand the highest quality science for acceptance, and to make MRD a home for important research that often has had trouble finding a home in other journals - such as research on less-famous model systems - all contributed to this outcome. Gary has been focused and tireless in this pursuit. Many of his efforts are invisible to authors or potential authors, yet have greatly increased the quality and rigor of what is considered by, and accepted in, MRD. Unique features like Gary's interesting front-matter articles (like his most-downloaded from 2012, below), special-issues, and the aesthetic VISIONS, also have been very special contributions that have made MRD an important and interesting Journal. Gary also brought in Julian Wong to be MRD's Managing Editor during the Wessel-years. Julian's attention to detail and his dedication to upgrading, automating, and streamlining the workflow at the Journal have been of great value to MRD's authors and editors. Each of us, individually, is grateful to Gary for the privilege to work with him, and to contribute as a team to the rise of the Journal. Gary's encouragement, ideas, and behind-the-scenes support delivered in thoughtful and personalized ways made it a joy to work with him on MRD. In keeping with Wiley Inc. policy that journals change Editors-in-Chief every 10 years, Gary is leaving the helm. However, an excellent successor waits in the wings, preparing to fill the big “shoes” that Gary leaves behind. Thanks, Gary! Harvey Florman Keith Latham Randy Prather Carmen Williams Mariana Wolfner

This Special Issue contains a collection of scientific and personal remembrances of Min Chueh (MC) Chang (October 10, 1908 – June 5, 1991). Chang is most reknowned today for three achievements: the co-discovery of sperm capacitation, his essential role in the development of the first generation oral contraceptive, and the development of methods for in vitro fertilization. The publication of this Special Issue collection is timed to celebrate his birth, 108 years ago this month; his passing, 25 years ago; his discovery of sperm capacitation 65 years ago (in 1951, see below); and 60 years since he, along with Gregory Pincus and collaborators, reported that orally administered progestins blocked ovulation in animals (Pincus et al., 1956). That 60-year anniversary publication, with a companion report describing the prevention of the human menstrual cycle by the same compounds (Garcia et al., 1956), was a landmark in the development of the Enovid contraceptive pill, and represents one measure of Chang’s impact within the long history of reproductive biology. To consider Chang’s contribution to our understanding of fertilization it is necessary to look back a century. The experimental study of sperm-egg interaction began in the middle of the 19 century with examination of external fertilization in amphibians (Newport, 1851; Newport, 1853; Newport and Ellis, 1854) and echinoderms (Hertwig, 1877; Fol, 1879). In these models, fertilization occurs rapidly and without specialized conditions: a cortical response occurs < 1 min after insemination in sea urchin eggs (Moser, 1939; Allen and Griffi n, 1958) while an oocyte’s fertilization potential is seen within several minutes in Xenopus (Webb and Nuccitelli, 1985; Kline et al., 1991). These tractable experimental models led to the development of key concepts in fertilization, including sperm chemoattraction by factors released from eggs (Lillie, 1912; Lillie, 1913a), specifi c cell adhesion events underlying spermegg interaction (Lillie, 1913b), and the block to polyspermy (Hertwig and Hertwig, 1887; Just, 1919). The mammalian egg was identifi ed in 1827 (von Baer and O’Malley, 1956) whereas sperm were known to exist much earlier from the work of Leeuwenhoek and of others (Cobb, 2007); the presence of sperm within the egg was observed by 1843 (Barry, 1843). Attempts to fertilize mammalian oocytes in vitro began at least as early as 1878 (Schenk, 1878; Onanoff, 1893; Long, 1912; Pincus, 1930; Pincus and Enzmann, 1934; Frommolt, 1934; Krasovkaja, 1935; Pincus, 1935; Yamane, 1935; Rock and Menkin, 1944; Menkin and Rock, 1948; Moricard, 1950; Venge, 1953; Shettles, 1953), but these were either not reproducible, not effi cient, or are questioned now due to the prior application of criteria for fertilization that are, today, viewed as insuffi ciently rigorous. (Another factor for this lack of success, not related to this discussion, is that the meiotic maturation of mammalian oocytes was also incompletely understood at the time.) Consequently, studies of mammalian fertilization remained largely descriptive until the middle of the 20 century. One reason for this stagnation was a failure to appreciate an unanticipated aspect of mammalian sperm function – namely capacitation. In 1951, Chang (Chang, 1951) and Colin Austin (Austin, 1951) independently reported that the ability of sperm to fertilize eggs is conferred by the environment of the female reproductive tract. This functional maturation, or “capacitation” (Austin, 1952), provided the perspective needed to develop reproducible methods to achieve mammalian fertilization in vitro. During the next few years, two groups, including Chang’s, unequivocally demonstrated the in vitro fertilization of rabbit eggs using ejaculated sperm recovered from the uterus (Dauzier et al., 1954; Chang, 1959), and hence had begun the process of capacitation in vivo (Yanagimachi, 1994). Importantly, Chang also showed that eggs fertilized in vitro can develop normally and produce live pups (Chang, 1959). In 1963, 85 years after the initial attempt by Schenk, Chang and Yanagimachi reported in vitro fertilization in the golden hamster using sperm recovered from the cauda epididymis and capacitated in vitro (Yanagimachi and Chang, 1963). By the end of the 1960s, in vitro fertilization was reported in mouse (Iwamatsu and Chang, 1969), human (Edwards et al., 1969), and several other mammals.

At the University of Massachusetts Medical School, anatomy, development, histology, and physiology teaching were integrated into a single first-year course where students study development, structure, and function in a context of clinical imaging. Although the integrated course comprised a 25 % reduction in teaching time, student course performance and USMLE Step I scores have remained strong since curricular integration and student satisfaction with teaching of these basic science disciplines in an interdisciplinary format is high.

Fertilization is a key step in the process of reproduction. In mammals, this begins with the entry of sperm into the fertile state by the process of capacitation, which occurs under instruction from the female reproductive tract, and is required for sperm access to and interaction with the egg. The cellular events of sperm–egg interaction result in gamete fusion and the initiation of the program of development and are further described in this chapter.

Molecular Reproduction and DevelopmentVolume 79, Issue 5 p. Fm i-Fm i EditorialFree Access Coup d'oeil of sperm Harvey M. Florman, Harvey M. Florman Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorWenlei Cao, Wenlei Cao Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorMelissa K. Jungnickel, Melissa K. Jungnickel Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorKeith A. Sutton, Keith A. Sutton Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorPei-Shiue Tsai, Pei-Shiue Tsai Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this author Harvey M. Florman, Harvey M. Florman Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorWenlei Cao, Wenlei Cao Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorMelissa K. Jungnickel, Melissa K. Jungnickel Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorKeith A. Sutton, Keith A. Sutton Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this authorPei-Shiue Tsai, Pei-Shiue Tsai Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655Search for more papers by this author First published: 16 April 2012 https://doi.org/10.1002/mrd.22039AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume79, Issue5May 2012Pages Fm i-Fm i RelatedInformation

The voltage-sensitive phosphoinositide phosphatases provide a mechanism to couple changes in the transmembrane electrical potential to intracellular signal transduction pathways. These proteins share a domain architecture that is conserved in deuterostomes. However, gene duplication events in primates, including humans, give rise to the paralogs TPTE and TPTE2 that retain protein domain organization but, in the case of TPTE, have lost catalytic activity. Here, we present evidence that these human proteins contain a functional voltage sensor, similar to that in nonmammalian orthologs. However, domains of these human proteins can also generate a noninactivating outward current that is not observed in zebra fish or tunicate orthologs. This outward current has the anticipated characteristics of a voltage-sensitive proton current and is due to the appearance of a single histidine residue in the S4 transmembrane segment of the voltage sensor. Histidine is observed at this position only during the eutherian radiation. Domains from both human paralogs generate proton currents. This apparent gain of proton channel function during the evolution of the TPTE protein family may account for the conservation of voltage sensor domains despite the loss of phosphatase activity in some human paralogs.