Aspects of spermatogenesis in immature and mature specimens of the long-lived Greenland shark: Novelties concerning the germinal compartment's assembly, complement of Sertoli cells and demise.

Cystic spermatogenesis in the subadult, maturing and adult Greenland shark (Somniosus microcephalus) displays multiple novel features, characterized early on by an unorganized internal cellular environment of the spermatocysts (anatomically discrete follicle-like units containing a single germ cell stage and its complement of co-developing Sertoli cells). These typically show polar asymmetries due to asymmetrically distributed germ and Sertoli cells. These arise from several novel cellular rearrangements at the immature pole, including fusion of a cluster of somatic cells with newly formed cysts containing only one to three spermatogonia and that already display an excess of Sertoli cells. The subadult's germinative zone revealed an additional novelty, namely numerous previously formed somatic cell-lined rings into which spermatogonia were incorporated. A striking finding was the conspicuous rarity of the routinely discernible Sertoli mitotic figures in the hallmark cyst stage of diametric elasmobranch spermatogenesis that is known for the peak display of the latter. Scrutiny of sequentially unfolding phenomena in the linearly arranged spermatogonial generations revealed that the cellular developments at the most common type of cyst-duct transition area (comprising slender to spindle-like basophilic cells with pointed ends) were concurrent with the discreet appearance of a second dark Sertoli nucleus, a development that persisted in spermiated cysts. Spermatogenically active mature males displayed vigorous meiotic divisions. However, a scattering of their spermatid cysts also displayed shark-atypical asynchronous passage through spermiogenesis, phenomena which were exacerbated as arrested spermiogenesis in an archival collection of tissues from 13 maturing specimens. Subadult specimens revealed meiotic arrest, and foci of infiltration of leukocytes that originate from a mass of eosinophilic, granule-laden immune cells dorsally under the testis capsule. This tissue was identical to the testis-affixed bone marrow equivalent in other shark species. This tissue is likely developmentally regulated in the Greenland shark as it is absent in adults.

Several routes of cell death to secondary necrosis in the elasmobranch testis.

The process of spermatogenesis features significant germ cell loss through apoptosis. Routine histology of the testes of well-studied animal models hardly discloses any trace of their phagocytic clearance by the supporting Sertoli cells. This review highlights lessons learnt from the cystic, diametric testes of some seasonally migrating elasmobranchs (e.g., spiny dogfish and blue sharks) that offer unconventional investigative paradigms to study these phenomena as these organs readily disclose a pronounced apoptosis gradient affecting exclusively spermatogonial clones that each are enclosed with their own Sertoli cells in spherical structures called spermatocysts. This gradient is visible at a certain time of year in the spermatogenically active shark, and peaks in mature spermatogonial cysts as clustered deaths with sporadic, and not massive secondary necrosis. Conversely, immature spermatogonial cysts in blue sharks reveal a characteristic periluminal display of single apoptotic deaths. Tracing aberrations in the immunostaining patterns of the conserved cell cycle marker, proliferating cell nuclear antigen, the gradual progression of the death process in individual or coalesced spermatogonia in contiguous cysts becomes clear. The multiple apoptotic nuclear fragmentation morphologies inform also of a protracted death process involving three different morphological routes of nuclear fragmentation (of which some are TUNEL-positive and other TUNEL-negative) and concomitant chromatin compaction that culminate in freed apoptotic bodies (i.e., secondary necrosis). It is discussed that the staggered spermatogonial deaths and accompanying intermittent secondary necrosis in mature blue shark spermatogonial cysts may well relate to the low phagocytosis capacity of cyst's Sertoli cells that are still functionally naïve.

Multiple Sources for Sertoli Cells and Two Sertoli Phenotypes in the Adult Elasmobranch Testis: Insight from Two Species Belonging to Different Orders.

Findings presented here for the chondrichthyans Prionace glauca and Isurus oxyrinchus show that the assembly of the spermatocyst and it's attached collecting duct are seamlessly connected developmental phenomena. The cyst's somatic cell component (i.e., Sertoli cells, SCs) and the duct's constituent cells derive both from a common precursor cell type (typically a large oblong cell) found among the A-spermatogonia in the folliculogenic region. Novel findings show that the co-developing collecting duct itself serves also as a source of either normal-looking or basophilic atypical SCs (aSCs), depending on whether the duct-cyst transition remains mitotically active and open, or is sealed. The aSCs arise from accumulating slender basophilic cells at the duct-cyst interface after which the newly formed cyst is sealed. Quantitative analysis of the latter in P. glauca revealed a correlation between the appearance of this aSC in immature cysts and the degenerated testicular condition that displays a gradient of multinucleate cell (MNC) death among spermatogonial cysts. Findings seem to implicate these aSCs in the life-death balance in mature spermatogonial cohorts downstream in the spermatogenic sequence, rather than in newly formed cysts that exhibit low rates of apoptosis. Photomicrographs of developing spermatogonial cysts showing several aSCs interspersed among cytoplasmically linked spermatogonia that are proliferating or have died, seem to suggest that these small SCs may be involved in confining MNC death to a given cyst region under conditions of subthreshold levels of apoptosis such as to ensure cyst recovery in immature spermatogonial cysts. Anat Rec, 301:1944-1954, 2018. © 2018 Wiley Periodicals, Inc.

The challenges of primary health care nurse leaders in the wake of New Health Care Reform in Norway.

BACKGROUND: The local municipality, whose management style is largely inspired by the New Public Management (NPM) model, has administrative responsibilities for primary health care in Norway. Those responsible for health care at the local level often find themselves torn between their professional responsibilities and the municipality's market-oriented funding system. The introduction of the new health care reform process known as the Coordination Reform in January 2012 prioritises primary health care while simultaneously promoting a more collaborative and multidisciplinary approach to health care. Nurse leaders experience constant cross-pressure in their roles as members of the municipal executive team, the execution of their professional and administrative duties, and the overall political aims of the new reform. The aim of this article is to illuminate some of the major challenges facing nurse leaders in charge of nursing homes and to draw attention to their professional concerns about the quality of nursing care with the introduction of the new reform and its implementation under NPM-inspired municipal executive leadership. METHOD: This study employs a qualitative design. In-depth interviews were conducted with 10 nurse leaders in 10 municipalities, with a phenomenological-hermeneutic approach used for data analysis and interpretation. RESULT: Findings highlighted the increasingly complex challenges facing nurse leaders operating in the context of the municipality's hierarchical NPM management structure, while they are required to exercise collaborative professional interactions as per the guidelines of the new Coordination Reform. The interview findings were interpreted out of three sub-themes 1) importance of support for the nurse leader, 2) concerns about overall service quality, and 3) increased tasks unrelated to nursing leadership. CONCLUSION: The priorities of municipal senior management and the focus of the municipality's care service need clarification in the light of this reform. The voices of those at the frontlines of the caring services need to be heard as the restructuring of the caring services may have implications both for funding allocation and for the quality of patient care.

A Comparative Study of Intratesticular Ductules in the Spermatogenically Active Testes of Shortfin Mako and Thresher Sharks.

This comparative study of the radial testes of sexually mature thresher sharks (Alopias vulpinus) and shortfin mako sharks (Isurus oxyrinchus) describes the histology of the three-tiered network of sperm-carrying ductules in the testis and the lymphomyeloid tissue associated with it, namely the epigonal organ. In both species, a testis → epigonal gradient was evident regarding the thickness of the ductule epithelial lining and subepithelial investment of connective tissue. Ductules straddling the testis-epigonal border often displayed luminal leukocytes and various signs of regression, including the progressive thickening of the ductule epithelial lining, dissolution of the cytoplasm, and loss of normal histoarchitecture. In Isurus, large amorphous areas formed due to the fusion of neighboring regressing ductules. The epigonal organ of Alopias additionally revealed circular degenerative sperm-containing, Hassall-like bodies with either a degenerate or cellular appearance, the latter the result of cell proliferative activity (as shown by proliferating cell nuclear antigen (PCNA) immunohistochemistry) in an expanding outer border comprising cells with intensely PCNA immunoreactive slender and oblong nuclei. The latter cells exhibited a periphery-to-center transformation of their nuclei, at which stage they were PCNA-negative and most likely in a terminally differentiated state as they phagocytized the cell debris in the degenerate core. Intermediate stages of these circular bodies were a rarity. The relationship between these degenerate bodies, and the common occurrence of blind pockets in the epithelial linings and non-apoptosis-related degenerate patches in the apical cytoplasmic regions of the irregular shaped ductules in Alopias is unclear, and needs further elucidation. Anat Rec, 299:1435-1448, 2016. © 2016 Wiley Periodicals, Inc.

Sperm of Galeorhinus galeus (Elasmobranchii, Triakidae) Traverse an Excurrent Duct System Characterized by Pronounced Regionalization: A Scanning Electron and Light Microscopy Study.

The transport and subsequent maturation of spermatozoa in the vertebrate excurrent duct require the creation of a series of biochemically defined luminal milieus along the length of the duct. Such specialization is accomplished, among others, by changes in the epididymal histoarchitecture. Here we show that the intratesticular and extratesticular genital ducts of mating Galeorhinus galeus exhibit pronounced regionalization both in terms of epithelial histology and lumen diameter size. Findings also reveal distinct differences in the manner in which the spermatozoa were found in each segment of the duct. Novel scanning electron microscopy evidence is presented showing that the wide lumen ductuli epididymides, which ultimately convey the spermatozoa to the proximal epididymis, show functional specialization as well. The wall of the former consisted of cuboidal ciliated and nonciliated cells whose spatial arrangement in the duct wall resulted in a luminal surface showing lengthy rows of cilia-free areas, with each row bordered on both sides by a single row of cilia. The proximal epididymis comprised several subregions whose epithelial histology varied widely. The distal epididymis and ampulla of the epididymis possessed many fingerlike projections and transverse septa, respectively. As the main storage site for spermatozoa, the ampulla completed the bundling of spermatozoa into spermatozeugmata. These were circular sperm masses in which the heads of the spermatozoa were aligned side by side and embedded in a seminal matrix, while their tails extended outward. These findings of pronounced regionalization differ greatly from the rather uniform epididymal histology seen in some rays.

The epigonal organ and mature pole of the testis in the recreationally fished blue shark (Prionace glauca): histochemico-functional correlates.

The exact role of the immune system in normal spermatogenesis is poorly understood. The attachment, however, of the lymphomyeloid epigonal organ specifically to the testis's mature pole in many shark species is a curious finding. Unlike the histology of the lymphomyeloid tissues of many other elasmobranchs, the epigonal organ leukocytes of wild-caught blue shark (Prionace glauca), besides exhibiting extensive nuclear heterogeneity, contain some of the largest known granules ever seen in vertebrate white blood cells. It was previously shown that the blue shark epigonal organ remains unremarkable and functionally unchanged despite cestode parasites embedded into its surface, suggesting that it might have other functions in addition to microbial defense. We show here that Prionace epigonal leukocytes shed their granule-laden cytoplasm into the cyst resorption zone (RZ) of the testis, i.e. the region separating the spermatogenic tissue from the epigonal organ, as they begin to migrate into the RZ. Using the immunoreactivity of the conserved transcription factor (proliferating cell nuclear antigen) as marker, it is shown that the granule-lacking leukocytes exclusively infiltrated spermatozoal cysts leftover after the wave of wide-spread multinuclear cell death in summer-breeding males in a seasonally dependent manner. By contrast, Prionace caught 2 months later showed fully recovered testes containing numerous completely intact spermatozoal cysts. Conversely, degenerating immature spermatids were gradually phagocytized by their accompanying Sertoli cells, and leukocytes did not infiltrate such cysts. The autoimmune response described here resembles in every aspect the testicular autoimmune response induced experimentally in a teleost fish. These observations suggest functional adaptation of shark leukocytes in response to specific changes in the testicular microenvironment.

Coordination of spermatogenic processes in the testis: lessons from cystic spermatogenesis.

A common observation in the vertebrate testis is that new germ cell clones enter spermatogenesis proper before previously formed clones have completed their development. The extent to which the developmental advance of any given germ cell clone in any phase of spermatogenesis is dependent on that of neighboring clones and/or on the coordinating influence of associated Sertoli cells in the immediate vicinity or of others further away remains unclear. This review presents an overall synthesis of findings in an ancient vertebrate, the spiny dogfish shark and shows that, even at this phyletic level, the developmental advance of a given germ cell clone is the outcome of various processes emanating from its spatiotemporal relationship with (1) its own complement of Sertoli cells in the anatomically distinct spermatocyst and (2) Sertoli cells associated with other germ cell clones that lie upstream or downstream in the spermatogenic progression and that secrete, among others, androgen and estrogen destined for target sites upstream. Analysis of the protracted spermatogenic cycle shows the coordination in space and time of spermatogenic and steroidogenic events. Furthermore, the natural withdrawal of pituitary gonadotropin support in the dogfish causes a distinct and highly ordered gradient of apoptosis among the spermatogonial generations; this in turn is a major contributing factor to the cyclic nature of sperm production observed in this lower vertebrate. Because of the simplicity of their testicular organization, their cystic spermatogenesis and their phylogenetic position, cartilaginous fishes constitute a valid vertebrate reference system for comparative analysis with higher vertebrates.

On making nursing undergraduate human reproductive physiology content meaningful and relevant: discussion of human pleasure in its biological context.

The traditional presentation of the Reproductive Physiology component in an Anatomy and Physiology course to nursing undergraduates focuses on the broad aspects of hormonal regulation of reproduction and gonadal anatomy, with the role of the higher centres of the brain omitted. An introductory discussion is proposed which could precede the lectures on the reproductive organs. The discussion gives an overview of the biological significance of human pleasure, the involvement of the neurotransmitter dopamine, and the role of pleasure in the survival of the individual and even species. Pleasure stimuli (positive and negative) and the biological significance of naturally-induced pleasurable experiences are briefly discussed in the context of reproduction and the preservation of genetic material with an aim to foster relevancy between subject material and human behaviour in any type of society. The tenderness of this aspect of the human existence is well-understood because of its invariable association with soul-revealing human expressions such as love, infatuation, sexual flirtations, all of which are underpinned by arousal, desire and/or pleasure. Assuming that increased knowledge correlates with increased confidence, the proposed approach may provide the nurse with an adequate knowledge base to overcome well-known barriers in communicating with their patients about matters of sexual health and intimacy.

Fetal bovine serum simultaneously stimulates apoptosis and DNA synthesis in premeiotic stages of spermatogenesis in spiny dogfish (Squalus acanthias) in vitro: modulation by androgen and spermatogenic activity status.

Using the simple cystic spermatogenesis in the shark testis as a model, we previously reported the relative resistance of immature spermatogonia (stem cell and early-stage spermatogonia) to apoptosis in the normal testis and after spermatoxicant exposure in vivo. Apoptosis was monitored by fluorescence image analysis of living cysts, using the validated acridine orange (AO) vital staining technique. Findings show that FBS simultaneously stimulates both apoptosis and [(3)H]thymidine incorporation in immature spermatogonial clones in a concentration-dependent manner in vitro. Furthermore, androgen inhibits apoptosis and increases cyst viability, more so with 10% FBS than with 1% FBS. All the effects were as a function of spermatogenic activity status but were distinct in early-stage spermatogonial cysts isolated from testes awakening from the previous winter spermatogenic arrest period. Results are discussed in the context of the alternating germ-Sertoli cell population kinetics of early-stage spermatogonial cysts in Squalus acanthias's protracted testicular cycle.