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Abstract The epididymal function and gene expression in mammals are under the control of the testis. Sex steroids are secreted from the testis and act on the epididymis in an endocrine manner. There is another, non-sex steroidal secreted signaling, named lumicrine signaling, in which testis-derived secreted proteins go through the male reproductive tract and act on the epididymis. The effects of such multiple regulations on the epididymis by the testis have been investigated for many genes. The recent development of high-throughput next-generation sequencing now enables us a further comparative survey of endocrine and lumicrine action-dependent gene expression. In the present study, testis-derived endocrine and lumicrine actions on epididymal gene expression were comparatively investigated by RNA-seq transcriptomic analyses. This investigation utilized experimental animal models in which testis-derived endocrine and/or lumicrine actions were interfered with, such as unilateral or bilateral orchidectomy. By bilateral orchidectomy, which interferes with both endocrine and lumicrine actions, 431 genes were downregulated. By unilateral orchidectomy, which also interferes with endocrine and lumicrine actions by the unilateral testis, but the endocrine action was compensated by the contralateral testis, 283 genes were downregulated. The content of such genes downregulated by unilateral orchidectomy was like those of lumicrine action-interfered efferent duct-ligation, W/Wv, and Nell2 −/− mice. When genes affected by unilateral and bilateral orchidectomy were compared, 154 genes were commonly downregulated, whereas 217 genes were specifically downregulated only by bilateral orchidectomy, indicating the distinction between endocrine and lumicrine actions on the proximal epididymal transcriptome. Comparative transcriptome analyses also showed that the expressions of genes emerging since Amniota were notably impacted by bilateral orchidectomy, unilateral orchidectomy, and lumicrine action-interfering treatments; the degree of influence from these treatments varied based on the evolutionary stage beyond Amniota. These findings unveil an evolutional transition of regulated gene expression in the proximal epididymis by two different testis-derived signaling mechanisms.

The physiological functions of the mammalian epididymis are typically regulated by the testes. In addition to sex steroids secreted by testicular Leydig cells, which act on the epididymis in an endocrine manner, there is a non-sex-steroidal signaling pathway known as the lumicrine pathway. This lumicrine signaling pathway involves ligand proteins secreted from germ cells within the testicular seminiferous tubules traversing the male reproductive tract, which induce epithelial differentiation in the epididymis. These findings prompted an inquiry into whether treatments influencing testis physiology can disrupt epididymal function by interfering with testis-epididymis communication. Busulfan, an alkylating agent commonly used to deplete testicular germ cells in reproductive biology, has not been sufficiently explored because of its effects on the epididymis. This study investigated the effects of busulfan administration on the proximal epididymis using histological and transcriptomic analyses. Notably, busulfan, as opposed to the vehicle dimethyl sulfoxide (DMSO), altered the morphology of the initial segment of the epididymis, leading to a reduction in the cell height of the luminal epithelium. RNA sequencing identified 185 significantly downregulated genes in the proximal epididymis of busulfan-administered mice compared to DMSO-administered mice. Comparative transcriptome analyses revealed similarities between the epididymal transcriptome of busulfan-administered mice and lumicrine-deficient mice, such as efferent-duct-ligated W/Wv and Nell2-/- mice. However, this differed from that of bilaterally orchidectomized mice, in which both the endocrine and lumicrine signaling pathways were simultaneously ablated. Collectively, these results suggested that the harmful effects of busulfan on the proximal epididymis are secondary consequences of the ablation of testis-epididymis lumicrine signaling.

The epididymal function and gene expression in mammals are under the control of the testis. Sex steroids are secreted from the testis and act on the epididymis in an endocrine manner. There is another, non-sex steroidal secreted signaling, named lumicrine signaling, in which testis-derived secreted proteins go through the male reproductive tract and act on the epididymis. The effects of such multiple regulations on the epididymis by the testis have been investigated for many genes. The recent development of high-throughput next-generation sequencing now enables us a further comparative survey of endocrine and lumicrine action-dependent gene expression. In the present study, testis-derived endocrine and lumicrine actions on epididymal gene expression were comparatively investigated by RNA-seq transcriptomic analyses. This investigation utilized experimental animal models in which testis-derived endocrine and/or lumicrine actions were interfered with, such as unilateral or bilateral orchidectomy. By bilateral orchidectomy, which interferes with both endocrine and lumicrine actions, 431 genes were downregulated. By unilateral orchidectomy, which also interferes with endocrine and lumicrine actions by the unilateral testis, but the endocrine action was compensated by the contralateral testis, 283 genes were downregulated. The content of such genes downregulated by unilateral orchidectomy was like those of lumicrine action-interfered efferent duct-ligation, W/Wv, and Nell2−/− mice. When genes affected by unilateral and bilateral orchidectomy were compared, 154 genes were commonly downregulated, whereas 217 genes were specifically downregulated only by bilateral orchidectomy, indicating the distinction between endocrine and lumicrine actions on the proximal epididymal transcriptome. Comparative transcriptome analyses also showed that the expressions of genes emerging since Amniota were notably impacted by bilateral orchidectomy, unilateral orchidectomy, and lumicrine action-interfering treatments; the degree of influence from these treatments varied based on the evolutionary stage beyond Amniota. These findings unveil an evolutional transition of regulated gene expression in the proximal epididymis by two different testis-derived signaling mechanisms. [ABSTRACT FROM AUTHOR]

Let G be an arbitrary group. For any subsets A and B of G, let A * B = { a * b ; a ∈ A , b ∈ B } where ' * ' is the binary operation on G. By μ G (r , s) , we denote the minimum cardinality of the set A * B , where | A | = r and | B | = s . In 2003, Eliahou et al. proposed a conjecture that for every finite group G of order g and every pair of integers r , s with 1 ≤ r , s ≤ g , k G (r , s) ≤ μ G (r , s) , where k G (r , s) = min d ∈ H (G) { (⌈ r d ⌉ + ⌈ s d ⌉ − 1) d } and H (G) is the set of orders of finite subgroups of G. In 2003, Eliahou et al. verified the conjecture for dihedral group D q of index q = p v . In this paper, we determine the upper bound for the size of μ Q 4 n (r , s) and μ U m , n (r , s) , where Q 4 n = 〈 a , b : a 2 n = e , b 2 = a n , a b = b a − 1 〉 is a dicyclic group of order 4n and U m , n = 〈 a , b : a 2 n = e , b m = e , a b a − 1 = b − 1 〉 and verify the conjecture proposed by Eliahou et al. [7] for Q 4 n for n to be a prime power. [ABSTRACT FROM AUTHOR]

Let G be an arbitrary group. For any subsets A and B of G, let A * B = { a * b ; a ∈ A , b ∈ B } where ' * ' is the binary operation on G. By μ G (r , s) , we denote the minimum cardinality of the set A * B , where | A | = r and | B | = s . In 2003, Eliahou et al. proposed a conjecture that for every finite group G of order g and every pair of integers r , s with 1 ≤ r , s ≤ g , k G (r , s) ≤ μ G (r , s) , where k G (r , s) = min d ∈ H (G) { (⌈ r d ⌉ + ⌈ s d ⌉ − 1) d } and H (G) is the set of orders of finite subgroups of G. In 2003, Eliahou et al. verified the conjecture for dihedral group D q of index q = p v . In this paper, we determine the upper bound for the size of μ Q 4 n (r , s) and μ U m , n (r , s) , where Q 4 n = 〈 a , b : a 2 n = e , b 2 = a n , a b = b a − 1 〉 is a dicyclic group of order 4n and U m , n = 〈 a , b : a 2 n = e , b m = e , a b a − 1 = b − 1 〉 and verify the conjecture proposed by Eliahou et al. [7] for Q 4 n for n to be a prime power. [ABSTRACT FROM AUTHOR]

Abstract Background Morphological and functional alterations in aging reproductive organs result in decreased male fertility. The epididymis functions as the transition region for post-testicular sperm maturation. And we have previously demonstrated that the epididymal initial segment (IS), a region of the reproductive tract essential for sperm maturation and capacitation, undergoes considerable histological changes and chronic immune activation in mice during aging. However, the local aging-associated cellular and molecular changes in the aged epididymal IS are poorly understood. Results We conducted single-cell RNA sequencing analysis on the epididymal IS of young (3-month-old) and old (21-month-old) mice. In total, 10,027 cells from the epididymal IS tissues of young and old mice were obtained and annotated. The cell composition, including the expansion of a principal cell subtype and Ms4a4b Hi Ms4a6b Hi T cells, changed with age. Aged principal cells displayed multiple functional gene expression changes associated with acrosome reaction and sperm maturation, suggesting an asynchronous process of sperm activation and maturation during epididymal transit. Meanwhile, aging-related altered pathways in immune cells, especially the “cell chemotaxis” in Cx3cr1 Hi epididymal dendritic cells (eDCs), were identified. The monocyte-specific expression of chemokine Ccl8 increased with age in eDCs. And the aged epididymal IS showed increased inflammatory cell infiltration and cytokine secretion. Furthermore, cell–cell communication analysis indicated that age increased inflammatory signaling in the epididymal IS. Conclusion Contrary to the general pattern of lower immune responses in the male proximal genital tract, we revealed an inflammaging status in mouse epididymal initial segment. These findings will allow future studies to enable the delay of male reproductive aging via immune regulation.

DIS3 is an RNA exosome associated ribonuclease that degrades a wide range of transcripts that can be essential for cell survival and development. The proximal region of the mouse epididymis (initial segment and caput) plays a pivotal role in sperm transport and maturation required for male fertility. However, whether DIS3 ribonuclease mediates RNA decay in proximal epididymides remains unclear. Herein, we established a conditional knockout mouse line by crossing a floxed Dis3 allele with Lcn9-cre mice in which the recombinase is expressed in the principal cells of initial segment as early as post-natal day 17. Morphological and histological analyses, immunofluorescence, computer-aided sperm analysis and fertility were used for functional analyses. We document that DIS3 deficiency in the initial segment had no effect on male fertility. Dis3 cKO males had normal spermatogenesis and initial segment development. In cauda epididymides of Dis3 cKO mice, sperm abundance, morphology, motility, and the frequency of acrosome exocytosis were comparable to controls. Collectively, our genetic model demonstrates that loss of DIS3 in the initial segment of the epididymis is not essential for sperm maturation, motility, or male fertility. • DIS3 ribonuclease is expressed in the mouse epididymal epithelium. • DIS3 ablation in the initial segment has no effect on male fertility. • Dis3 cKO males have normal spermatogenesis and initial segment development. • Dis3 cKO males have normal sperm abundance, morphology, motility and acrosome reaction. [ABSTRACT FROM AUTHOR]

Background: The proximal region of the mouse epididymis plays a pivotal role in sperm transport, sperm maturation, and male fertility. Several studies have focused on segment-dependent gene expression of the mouse epididymis through high-throughput sequencing without the precision of the microdissection. Methods and results: Herein, we isolated the initial segment (IS) and proximal caput (P-caput) by physical microdissection using an Lcn9-cre; Rosa26tdTomato mouse model. We defined the transcriptome changes of caput epididymis by RNA sequencing (RNA-seq), which identified 1,961 genes that were abundantly expressed in the IS and 1,739 genes that were prominently expressed in the P-caput. In addition, we found that many differentially expressed genes (DEGs) were predominantly or uniquely expressed in the epididymis and region-specific genes were highly associated with transport, secretion, sperm motility, fertilization, and male fertility. Conclusion: Thus, this study provides an RNA-seq resource to identify regionspecific genes in the caput epididymis. The epididymal-selective/specific genes are potential targets for male contraception and may provide new insights into understanding segment-specific epididymal microenvironment-mediated sperm transport, maturation, and male fertility. [ABSTRACT FROM AUTHOR]

Background: Morphological and functional alterations in aging reproductive organs result in decreased male fertility. The epididymis functions as the transition region for post-testicular sperm maturation. And we have previously demonstrated that the epididymal initial segment (IS), a region of the reproductive tract essential for sperm maturation and capacitation, undergoes considerable histological changes and chronic immune activation in mice during aging. However, the local aging-associated cellular and molecular changes in the aged epididymal IS are poorly understood. Results: We conducted single-cell RNA sequencing analysis on the epididymal IS of young (3-month-old) and old (21-month-old) mice. In total, 10,027 cells from the epididymal IS tissues of young and old mice were obtained and annotated. The cell composition, including the expansion of a principal cell subtype and Ms4a4bHiMs4a6bHi T cells, changed with age. Aged principal cells displayed multiple functional gene expression changes associated with acrosome reaction and sperm maturation, suggesting an asynchronous process of sperm activation and maturation during epididymal transit. Meanwhile, aging-related altered pathways in immune cells, especially the "cell chemotaxis" in Cx3cr1Hi epididymal dendritic cells (eDCs), were identified. The monocyte-specific expression of chemokine Ccl8 increased with age in eDCs. And the aged epididymal IS showed increased inflammatory cell infiltration and cytokine secretion. Furthermore, cell–cell communication analysis indicated that age increased inflammatory signaling in the epididymal IS. Conclusion: Contrary to the general pattern of lower immune responses in the male proximal genital tract, we revealed an inflammaging status in mouse epididymal initial segment. These findings will allow future studies to enable the delay of male reproductive aging via immune regulation. [ABSTRACT FROM AUTHOR]

EXOSC10 is an exosome-associated ribonuclease that degrades and processes a wide range of transcripts in the nucleus. The initial segment (IS) of the epididymis is crucial for sperm transport and maturation in mice by affecting the absorption and secretion that is required for male fertility. However, the role of EXOSC10 ribonuclease-mediated RNA metabolism within the IS in the regulation of gene expression and sperm maturation remains unknown. Herein, we established an Exosc10 conditional knockout ( Exosc10 cKO) mouse model by crossing Exosc10 F/F mice with Lcn9-Cre mice which expressed recombinase in the principal cells of IS as early as post-natal day 17. Morphological and histological analyses revealed that Exosc10 cKO males had normal spermatogenesis and development of IS. Moreover, the sperm concentration, morphology, motility, and frequency of acrosome reactions in the cauda epididymides of Exosc10 cKO mice were comparable with those of control mice. Thus, Exosc10 cKO males had normal fertility. Collectively, our genetic mouse model and findings demonstrate that loss of EXOSC10 in the IS of epididymis is dispensable for sperm maturation and male fertility.

It is known that every non-universal self-full degree in the structure of the degrees of computably enumerable equivalence relations (ceers) under computable reducibility has exactly one strong minimal cover. This leaves little room for embedding wide partial orders as initial segments using self-full degrees. We show that considerably more can be done by staying entirely inside the collection of non-self-full degrees. We show that the poset can be embedded as an initial segment of the degrees of ceers with infinitely many classes. A further refinement of the proof shows that one can also embed the free distributive lattice generated by the lower semilattice as an initial segment of the degrees of ceers with infinitely many classes. [ABSTRACT FROM AUTHOR]

DIS3L, a catalytic exoribonuclease associated with the cytoplasmic exosome complex, degrades cytoplasmic RNAs and is implicated in cancers and certain other diseases in humans. Epididymis plays a pivotal role in the transport, maturation, and storage of sperm required for male fertility. However, it remains unclear whether DIS3L-mediated cytoplasmic RNA degradation plays a role in epididymis biology and functioning. Herein, we fabricated a Dis3l conditional knockout (Dis3l cKO) mouse line in which DIS3L was ablated from the principal cells of the initial segment (IS). Morphological analyses showed that spermatogenesis and IS differentiation occurred normally in Dis3l cKO mice. Additionally, the absence of DIS3L had no dramatic influence on the transcriptome of IS. Moreover, the sperm count, morphology, motility, and acrosome reaction frequency in Dis3l cKO mice were comparable to that of the control, indicating that the Dis3l cKO males had normal fertility. Collectively, our genetic model demonstrates that DIS3L inactivation in the IS is nonessential for sperm maturation and male fertility., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest regarding the publication of this research., (Copyright © 2024 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

Ｓｅｄｉｍｅｎｔ ｉｓ ｃａｒｒｉｅｄ ｂｙ ｆｌｏｗ ｔｏ ｔｈｅ ｌａｋｅ ａｒｅａ ｄｕｒｉｎｇ ｔｈｅ ｆｏｒｍａｔｉｏｎ ｐｒｏｃｅｓｓ ｏｆ ｔｈｅ ｉｎｉ⁃ ｔｉａｌ ｓｅｇｍｅｎｔ ｏｆ ａ ｄｅｌｔａ， ｗｈｉｃｈ ｍａｉｎｌｙ ｄｅｐｅｎｄｓ ｏｎ ｔｈｅ ｉｎｉｔｉａｌ ｍｏｍｅｎｔｕｍ ｔｏ ｍａｉｎｔａｉｎ ｉｔｓ ｏｗｎ ｃｏｎ⁃ ｔｉｎｕｏｕｓ ｍｏｖｅｍｅｎｔ． Ａｃｃｏｒｄｉｎｇ ｔｏ ｔｈｅ ｃｈａｒａｃｔｅｒｉｓｔｉｃｓ ｏｆ ｔｈｉｓ ｐｒｏｃｅｓｓ， ｔｈｅ ｔｈｅｏｒｅｔｉｃａｌ ｍｏｄｅｌ ｆｏｒ ｔｈｅ ｐｌａｎｅ ｊｅｔ ｂｏｕｎｄａｒｙ ｌａｙｅｒ ｉｎ ｔｈｅ ｉｎｉｔｉａｌ ｓｅｇｍｅｎｔ ｏｆ ｔｈｅ ｄｅｌｔａ ｗａｓ ｅｓｔａｂｌｉｓｈｅｄ ｂａｓｅｄ ｏｎ ｔｈｅ ｓｈａｌｌｏｗ ｗａｔｅｒ ｅｑｕａｔｉｏｎｓ ｆｏｒ ｍｕｄｄｙ ｗａｔｅｒ， ａｎｄ ｔｈｅ ｆｌｏｗ ｆｉｅｌｄ ｄｉｓｔｒｉｂｕｔｉｏｎ ｏｆ ｔｈｅ ｉｎｉｔｉａｌ ｓｅｇｍｅｎｔ ｏｆ ｄｅｌｔａ ｆｏｒｍａｔｉｏｎ ｗａｓ ｏｂｔａｉｎｅｄ ｗｉｔｈ ｔｈｅ ｓｉｍｉｌａｒｉｔｙ ｓｏｌｕｔｉｏｎ ｍｅｔｈｏｄ． Ｂａｓｅｄ ｏｎ ｔｈｅ ｇｅｎｅｒａｌ ｍａｔｈ⁃ ｅｍａｔｉｃａｌ ｍｏｄｅｌ ｏｆ ｒｉｖｅｒ ｂｅｄ ｅｖｏｌｕｔｉｏｎ， ｔｈｅ ｔｈｅｏｒｅｔｉｃａｌ ｅｘｐｒｅｓｓｉｏｎ ｏｆ ｔｈｅ ｍｏｒｐｈｏｌｏｇｉｃａｌ ｃｈａｒａｃ⁃ ｔｅｒｉｓｔｉｃｓ ｏｆ ｔｈｅ ｉｎｉｔｉａｌ ｓｅｇｍｅｎｔ ｗａｓ ｄｅｒｉｖｅｄ， ａｎｄ ｔｈｅ ｅｒｏｓｉｏｎ ａｎｄ ｄｅｐｏｓｉｔｉｏｎ ｏｆ ｔｈｅ ｉｎｉｔｉａｌ ｓｅｇｍｅｎｔ ｗｅｒｅ ｑｕａｎｔｉｔａｔｉｖｅｌｙ ａｎａｌｙｚｅｄ． Ｔｈｒｏｕｇｈ ｔｈｅ ｅｘｐｅｒｉｍｅｎｔａｌ ｖｅｒｉｆｉｃａｔｉｏｎ， ｔｈｅ ｔｈｅｏｒｅｔｉｃａｌ ｓｏｌｕｔｉｏｎ ｃａｎ ｗｅｌｌ ｄｅｓｃｒｉｂｅ ｔｈｅ ｅｒｏｓｉｏｎ ａｎｄ ｄｅｐｏｓｉｔｉｏｎ ｔｒｅｎｄ ａｎｄ ｍｏｒｐｈｏｌｏｇｉｃａｌ ｃｈａｒａｃｔｅｒｉｓｔｉｃｓ ｉｎ ｔｈｅ ｅａｒｌｙ ｓｔａｇｅ ｏｆ ｄｅｌｔａ ｆｏｒｍａｔｉｏｎ． [ABSTRACT FROM AUTHOR]

Action potentials (APs) in nigral dopaminergic neurons often exhibit two separate components: the first reflecting spike initiation in the dendritically located axon initial segment (AIS) and the second the subsequent dendro-somatic spike. These components are separated by a notch in the ascending phase of the somatic extracellular waveform and in the temporal derivative of the somatic intracellular waveform. Still, considerable variability exists in the presence and magnitude of the notch across neurons. To systematically address the contribution of AIS, dendritic and somatic compartments to shaping the two-component APs, we modeled APs of previously in vivo electrophysiologically characterized and 3D-reconstructed male mouse and rat dopaminergic neurons. A parsimonious two-domain model, with high (AIS) and lower (dendro-somatic) Na+ conductance, reproduced the notch in the temporal derivatives, but not in the extracellular APs, regardless of morphology. The notch was only revealed when somatic active currents were reduced, constraining the model to three domains. Thus, an initial AIS spike is followed by an actively generated spike by the axon-bearing dendrite (ABD), in turn followed mostly passively by the soma. The transition from being a source compartment for the AIS spike to a source compartment for the ABD spike satisfactorily explains the extracellular somatic notch. Larger AISs and thinner ABD (but not soma-to-AIS distance) accentuate the AIS component. We conclude that variability in AIS size and ABD caliber explains variability in AP extracellular waveform and separation of AIS and dendro-somatic components, given the presence of at least three functional domains with distinct excitability characteristics. [ABSTRACT FROM AUTHOR]