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A temporal and spatial map of axons in developing mouse prostate

  • Anne E. Turco
  • Mark T. Cadena
  • Helen L. Zhang
  • Jaskiran K. Sandhu
  • Steven R. Oakes
  • Thrishna Chathurvedula
  • Richard E. Peterson
  • Janet R. Keast
  • Chad M. VezinaEmail author
Original Paper

Abstract

Prostate autonomic and sensory axons control glandular growth, fluid secretion, and smooth muscle contraction and are remodeled during cancer and inflammation. Morphogenetic signaling pathways reawakened during disease progression may drive this axon remodeling. These pathways are linked to proliferative activities in prostate cancer and benign prostate hyperplasia. However, little is known about which developmental signaling pathways guide axon investment into prostate. The first step in defining these pathways is pinpointing when axon subtypes first appear in prostate. We accomplished this by immunohistochemically mapping three axon subtypes (noradrenergic, cholinergic, and peptidergic) during fetal, neonatal, and adult stages of mouse prostate development. We devised a method for peri-prostatic axon density quantification and tested whether innervation is uniform across the proximo–distal axis of dorsal and ventral adult mouse prostate. Many axons directly interact with or innervate neuroendocrine cells in other organs, so we examined whether sensory or autonomic axons innervate neuroendocrine cells in prostate. We first detected noradrenergic, cholinergic, and peptidergic axons in prostate at embryonic day (E) 14.5. Noradrenergic and cholinergic axon densities are uniform across the proximal–distal axis of adult mouse prostate while peptidergic axons are denser in the periurethral and proximal regions. Peptidergic and cholinergic axons are closely associated with prostate neuroendocrine cells whereas noradrenergic axons are not. These results provide a foundation for understanding mouse prostatic axon development and organization and, provide strategies for quantifying axons during progression of prostate disease.

Keywords

Mouse Axon development Prostate innervation Sympathetic Parasympathetic 

Notes

Acknowledgements

This work was supported by National Institutes of Health Grants RO1ES001332, U54DK104310, T32ES007015, U01DK110807, and U01DK110807-S1.

Compliance with ethical standards

Conflict of Interest

The authors report no conflict of interest.

Supplementary material

418_2019_1784_MOESM1_ESM.tif (23.3 mb)
Supplementary Fig. 1 CGRP+, VAChT+, and TH+ axons are denser during prostate branching morphogenesis and sexual maturity than during prostatic bud formation. E17.5, P9, and P50 mouse prostate tissue sections were immunostained with antibodies against TH+, VAChT+, and CGRP+ (green) and e-cadherin (CDH1, to visualize prostatic and urethral epithelium, red). Axon pixel densities were quantified in the 10 μM periductal spaces radiating outward from the basilar surface of prostatic epithelium and results are shown pictorially on the right. Yellow arrowheads identify varicose axons. Results are mean ± SE of seven mice per group and three non-adjacent tissue sections per mouse. Asterisks indicate significant differences (*p < 0.05, and **p < 0.01) between regions. Scale bar is 50 µm (TIFF 23808 kb)
418_2019_1784_MOESM2_ESM.tif (26.6 mb)
Supplementary Fig. 2 CGRP+ axon density in P50 mouse ventral prostate is greater in the periurethral and proximal region than the distal region while VAChT+ and TH+ axon density does not significantly differ between proximal–distal regions. P50 mouse prostate tissue sections were immunostained with antibodies against TH+, VAChT+, or CGRP+ (green) and e-cadherin (CDH1, to visualize prostatic and urethral epithelium, red). Regions selected for analysis are schematized at the top of the figure and representative images are shown at the bottom left. Axon pixel densities were quantified in the 10 μM periductal spaces radiating outward from the basal surface of prostatic epithelium and results are shown pictorially at the right. Results are mean ± SE of ten mice per group and three non-adjacent tissue sections per mouse. Asterisks indicate significant differences (**p < 0.01) between regions. Scale bar is 50 µm (TIFF 27271 kb)
418_2019_1784_MOESM3_ESM.tif (4.4 mb)
Supplementary Fig. 3 Graphs showing CGRP+ axon density in P50 mouse dorsal prostate is greater in the periurethral and proximal region than the distal region while VAChT+ and TH+ axon density does not significantly differ between proximal–distal regions. Axon densities in P50 mouse prostate tissue sections were determined using the procedure described in Methods and Fig. 1 legend. Axon pixel densities were quantified in the 10 μm periductal spaces radiating outward from the basal surface of prostatic epithelium and results graphed. (a) CGRP+ axon density in the periurethral and proximal region is significantly greater than in the distal region. (b) VAChT+ axon density is uniform across the periurethral, proximal, and distal regions. (c) TH+ axon density is uniform across the periurethral, proximal, and distal regions. Results are mean ± SE of ten mice per group and three non-adjacent tissue sections per mouse. Asterisks indicate significant differences (**p < 0.01) between regions. RS identifies the rhabdosphincter. Scale bar is 50 µm (TIFF 4497 kb)
418_2019_1784_MOESM4_ESM.tif (2.2 mb)
Supplementary Fig. 4 Graphs showing CGRP+ axon density in P50 mouse ventral prostate is greater in the periurethral and proximal region than the distal region while VAChT+ and TH+ axon density does not significantly differ between proximal–distal regions. Axon densities in P50 mouse prostate tissue sections were determined using the procedure described in Methods and Fig. 1 legend. Axon pixel densities were quantified in the 10 μm periductal spaces radiating outward from the basal surface of prostatic epithelium and results graphed. (a) The CGRP+ axon density in the periurethral and proximal region is significantly greater than in the distal region. (b) VAChT+ axon density is uniform across the periurethral, proximal and distal regions. (c) TH+ axon density is uniform across the periurethral, proximal and distal regions. Results are mean ± SE of ten mice per group and three non-adjacent tissue sections per mouse. Asterisks indicate significant differences (*p < 0.05, and **p < 0.01) between regions (TIFF 2233 kb)
418_2019_1784_MOESM5_ESM.docx (32 kb)
Supplementary Table 1 List of primary antibodies used in this study. The antibody target, vendor and catelog number, RRID number, host species, dilution, and proof of specificity is listed. (DOCX 31 kb)

References

  1. Abler LL, Keil KP, Mehta V, Joshi PS, Schmitz CT, Vezina CM (2011) A high-resolution molecular atlas of the fetal mouse lower urogenital tract. Dev Dyn 240(10):2364–2377.  https://doi.org/10.1002/dvdy.22730 CrossRefGoogle Scholar
  2. Aven L, Ai X (2013) Mechanisms of respiratory innervation during embryonic development. Organogenesis 9(3):194–198.  https://doi.org/10.4161/org.24842 CrossRefGoogle Scholar
  3. Batra S, Christensson PI, Hartley-Asp B (1990) Characterization of muscarinic cholinergic receptors in membrane preparations from rat prostatic adenocarcinoma. Prostate 17(4):261–268.  https://doi.org/10.1002/pros.2990170402 CrossRefGoogle Scholar
  4. Baumgarten HG, Falck B, Holstein AF, Owman C, Owman T (1968) Adrenergic innervation of the human testis, epididymis, ductus deferens and prostate: a fluorescence microscopic and fluorimetric study. Z Zellforsch Mikrosk Anat 90(1):81–95CrossRefGoogle Scholar
  5. Belle M, Godefroy D, Couly G, Malone SA, Collier F, Giacobini P, Chédotal A (2017) Tridimensional visualization and analysis of early human development. Cell 169(1):161–173.e12.  https://doi.org/10.1016/j.cell.2017.03.008 CrossRefGoogle Scholar
  6. Bianchi-Frias D, Vakar-Lopez F, Coleman IM, Plymate SR, Reed MJ, Nelson PS (2010) The effects of aging on the molecular and cellular composition of the prostate microenvironment. PLoS One 5(9):e12501.  https://doi.org/10.1371/journal.pone.0012501 CrossRefGoogle Scholar
  7. Branchfield K, Nantie L, Verheyden JM, Sui P, Wienhold MD, Sun X (2016) Pulmonary neuroendocrine cells function as airway sensors to control lung immune response. Science 351(6274):707–710.  https://doi.org/10.1126/science.aad7969 CrossRefGoogle Scholar
  8. Caine M, Raz S, Zeigler M (1975) Adrenergic and cholinergic receptors in the human prostate, prostatic capsule and bladder neck. Br J Urol 47(2):193–202CrossRefGoogle Scholar
  9. Cheng C-Y, Zhou Z, Nikitin AY (2013) Detection and organ-specific ablation of neuroendocrine cells by synaptophysin locus-based BAC cassette in transgenic mice. PLoS One 8(4):e60905.  https://doi.org/10.1371/journal.pone.0060905 CrossRefGoogle Scholar
  10. Danziger ZC, Grill WM (2017) Sensory feedback from the urethra evokes state-dependent lower urinary tract reflexes in rat. J Physiol (Lond). 595(16):5687–5698.  https://doi.org/10.1113/JP274191 CrossRefGoogle Scholar
  11. de Groat WC, Yoshimura N (2009) Afferent nerve regulation of bladder function in health and disease. Handb Exp Pharmacol 194:91–138.  https://doi.org/10.1007/978-3-540-79090-7_4 CrossRefGoogle Scholar
  12. Farrell J, Lyman Y (1937) A study of the secretory nerves of, and the action of certain drugs on the prostate gland. Am J Physiol 118:64–70CrossRefGoogle Scholar
  13. Georgas KM, Armstrong J, Keast JR, Larkins CE, McHugh KM, Southard-Smith EM, Cohn MJ, Batourina E, Dan H, Schneider K et al (2015) An illustrated anatomical ontology of the developing mouse lower urogenital tract. Development 142(10):1893–1908.  https://doi.org/10.1242/dev.117903 CrossRefGoogle Scholar
  14. Harburg GC, Hinck L (2011) Navigating breast cancer: axon guidance molecules as breast cancer tumor suppressors and oncogenes. J Mammary Gland Biol Neoplasia 16(3):257–270.  https://doi.org/10.1007/s10911-011-9225-1 CrossRefGoogle Scholar
  15. Harding SD, Armit C, Armstrong J, Brennan J, Cheng Y, Haggarty B, Houghton D, Lloyd-MacGilp S, Pi X, Roochun Y et al (2011) The GUDMAP database—an online resource for genitourinary research. Development. 138(13):2845–2853.  https://doi.org/10.1242/dev.063594 CrossRefGoogle Scholar
  16. Hatch J, Mukouyama Y-S (2015) Spatiotemporal mapping of vascularization and innervation in the fetal murine intestine. Dev Dyn 244(1):56–68.  https://doi.org/10.1002/dvdy.24178 CrossRefGoogle Scholar
  17. Kapur J, Sahoo P, Wong A (1985) A new method for gray-level picture thresholding using the entropy of the histogram. Graph Models Image Process 29(3):273–285CrossRefGoogle Scholar
  18. Keast JR, Smith-Anttila CJA, Osborne PB (2015) Developing a functional urinary bladder: a neuronal context. Front Cell Dev Biol 3.  https://doi.org/10.3389/fcell.2015.00053. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4555086/. Accessed 4 Jun 2018
  19. Keil KP, Mehta V, Abler LL, Joshi PS, Schmitz CT, Vezina CM (2012) Visualization and quantification of mouse prostate development by in situ hybridization. Differentiation 84(3):232–239.  https://doi.org/10.1016/j.diff.2012.07.005 CrossRefGoogle Scholar
  20. Kepper M, Keast J (1995) Immunohistochemical properties and spinal connections of pelvic autonomic neurons that innervate the rat prostate gland. Cell Tissue Res 281(3):533–542CrossRefGoogle Scholar
  21. Kopp UC (2015) Role of renal sensory nerves in physiological and pathophysiological conditions. Am J Physiol Regul Integr Comp Physiol 308(2):R79–R95.  https://doi.org/10.1152/ajpregu.00351.2014 CrossRefGoogle Scholar
  22. Kummer W, Lips KS, Pfeil U (2008) The epithelial cholinergic system of the airways. Histochem Cell Biol 130(2):219–234.  https://doi.org/10.1007/s00418-008-0455-2 CrossRefGoogle Scholar
  23. Lamb JP, Sparrow MP (2002) Three-dimensional mapping of sensory innervation with substance p in porcine bronchial mucosa. Am J Respir Crit Care Med 166(9):1269–1281.  https://doi.org/10.1164/rccm.2112018 CrossRefGoogle Scholar
  24. Lanlua P, Decorti F, Gangula PR, Chung K, Taglialatela G, Yallampalli C (2001) Female steroid hormones modulate receptors for nerve growth factor in rat dorsal root ganglia. Biol Reprod 64(1):331–338CrossRefGoogle Scholar
  25. Lin T-M, Rasmussen NT, Moore RW, Albrecht RM, Peterson RE (2003) Region-specific inhibition of prostatic epithelial bud formation in the urogenital sinus of C57BL/6 mice exposed in utero to 2,3,7,8-tetrachlorodibenzo-p-dioxin. Toxicol Sci 76(1):171–181.  https://doi.org/10.1093/toxsci/kfg218 CrossRefGoogle Scholar
  26. Magnon C, Hall SJ, Lin J, Xue X, Gerber L, Freedland SJ, Frenette PS (2013) Autonomic nerve development contributes to prostate cancer progression. Science 341(6142):1236361.  https://doi.org/10.1126/science.1236361 CrossRefGoogle Scholar
  27. Marker PC, Donjacour AA, Dahiya R, Cunha GR (2003) Hormonal, cellular, and molecular control of prostatic development. Dev Biol 253(2):165–174CrossRefGoogle Scholar
  28. McVary KT, Razzaq A, Lee C, Venegas MF, Rademaker A, McKenna KE (1994) Growth of the rat prostate gland is facilitated by the autonomic nervous system. Biol Reprod 51(1):99–107CrossRefGoogle Scholar
  29. Nassenstein C, Kwong K, Taylor-Clark T, Kollarik M, MacGlashan DM, Braun A, Undem BJ (2008) Expression and function of the ion channel TRPA1 in vagal afferent nerves innervating mouse lungs. J Physiol 586(Pt 6):1595–1604.  https://doi.org/10.1113/jphysiol.2007.148379 CrossRefGoogle Scholar
  30. Nicholson TM, Ricke WA (2011) Androgens and estrogens in benign prostatic hyperplasia: past, present and future. Differentiation 82(4–5):184–199.  https://doi.org/10.1016/j.diff.2011.04.006 CrossRefGoogle Scholar
  31. Oliveira DSM, Dzinic S, Bonfil AI, Saliganan AD, Sheng S, Bonfil RD (2016) The mouse prostate: a basic anatomical and histological guideline. Bosn J Basic Med Sci 16(1):8–13.  https://doi.org/10.17305/bjbms.2016.917 Google Scholar
  32. Owman C, Sjöstrand NO (1965) Short adrenergic neurons and catecholamine-containing cells in vas deferens and accessory male genital glands of different mammals. Zeitschrift für Zellforschung 66(2):300–320.  https://doi.org/10.1007/BF00344342 CrossRefGoogle Scholar
  33. Pennefather JN, Lau WA, Mitchelson F, Ventura S (2000) The autonomic and sensory innervation of the smooth muscle of the prostate gland: a review of pharmacological and histological studies. J Auton Pharmacol 20(4):193–206CrossRefGoogle Scholar
  34. Pflug BR, Onoda M, Lynch JH, Djakiew D (1992) Reduced expression of the low affinity nerve growth factor receptor in benign and malignant human prostate tissue and loss of expression in four human metastatic prostate tumor cell lines. Cancer Res 52(19):5403–5406Google Scholar
  35. Pontari MA, Ruggieri MR (2004) Mechanisms in prostatitis/chronic pelvic pain syndrome. J Urol 172(3):839–845.  https://doi.org/10.1097/01.ju.0000136002.76898.04 CrossRefGoogle Scholar
  36. Premkumar LS (2014) Transient receptor potential channels as targets for phytochemicals. ACS Chem Neurosci 5(11):1117–1130.  https://doi.org/10.1021/cn500094a CrossRefGoogle Scholar
  37. Raz S, Zeigler M, Caine M (1973) The effect of progesterone on the adrenergic receptors of the urethra1. Br J Urol 45(2):131–135.  https://doi.org/10.1111/j.1464-410X.1973.tb12129.x CrossRefGoogle Scholar
  38. Sjöstrand NO (1965) The adrenergic innervation of the vas deferens and the accessory male genital glands. Acta Physiol Scand (Suppl):1–82Google Scholar
  39. Szczyrba J, Niesen A, Wagner M, Wandernoth PM, Aumüller G, Wennemuth G (2017) Neuroendocrine cells of the prostate derive from the neural crest. J Biol Chem 292(5):2021–2031.  https://doi.org/10.1074/jbc.M116.755082 CrossRefGoogle Scholar
  40. Thomson AA, Timms BG, Barton L, Cunha GR, Grace OC (2002) The role of smooth muscle in regulating prostatic induction. Development 129(8):1905–1912Google Scholar
  41. Toivanen R, Shen MM (2017) Prostate organogenesis: tissue induction, hormonal regulation and cell type specification. Development 144(8):1382–1398.  https://doi.org/10.1242/dev.148270 CrossRefGoogle Scholar
  42. Tollet J, Everett AW, Sparrow MP (2001) Spatial and temporal distribution of nerves, ganglia, and smooth muscle during the early pseudoglandular stage of fetal mouse lung development. Dev Dyn 221(1):48–60.  https://doi.org/10.1002/dvdy.1124 CrossRefGoogle Scholar
  43. Umans BD, Liberles SD (2018) Neural sensing of organ volume. Trends Neurosci 15:85.  https://doi.org/10.1016/j.tins.2018.07.008 Google Scholar
  44. Ventura S, Pennefather J, Mitchelson F (2002) Cholinergic innervation and function in the prostate gland. Pharmacol Ther 94(1–2):93–112CrossRefGoogle Scholar
  45. Vezina CM, Allgeier SH, Moore RW, Lin T-M, Bemis JC, Hardin HA, Gasiewicz TA, Peterson RE (2008) Dioxin causes ventral prostate agenesis by disrupting dorsoventral patterning in developing mouse prostate. Toxicol Sci 106(2):488–496.  https://doi.org/10.1093/toxsci/kfn183 CrossRefGoogle Scholar
  46. Wang JM, McKenna KE, McVary KT, Lee C (1991) Requirement of innervation for maintenance of structural and functional integrity in the rat prostate. Biol Reprod 44(6):1171–1176CrossRefGoogle Scholar
  47. Watanabe T, Inoue M, Sasaki K, Araki M, Uehara S, Monden K, Saika T, Nasu Y, Kumon H, Chancellor MB (2011) Nerve growth factor level in the prostatic fluid of patients with chronic prostatitis/chronic pelvic pain syndrome is correlated with symptom severity and response to treatment. BJU Int 108(2):248–251.  https://doi.org/10.1111/j.1464-410X.2010.09716.x CrossRefGoogle Scholar
  48. White CW, Xie JH, Ventura S (2013) Age-related changes in the innervation of the prostate gland. Organogenesis 9(3):206–215.  https://doi.org/10.4161/org.24843 CrossRefGoogle Scholar
  49. Yan H, Keast JR (2008) Neurturin regulates postnatal differentiation of parasympathetic pelvic ganglion neurons, initial axonal projections, and maintenance of terminal fields in male urogenital organs. J Comp Neurol 507(2):1169–1183.  https://doi.org/10.1002/cne.21593 CrossRefGoogle Scholar
  50. Zahalka AH, Arnal-Estapé A, Maryanovich M, Nakahara F, Cruz CD, Finley LWS, Frenette PS (2017) Adrenergic nerves activate an angio-metabolic switch in prostate cancer. Science 358(6361):321–326.  https://doi.org/10.1126/science.aah5072 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Molecular and Environmental Toxicology Center, University of Wisconsin-MadisonMadisonUSA
  2. 2.Pharmaceutical Sciences Division, School of PharmacyUniversity of Wisconsin-MadisonMadisonUSA
  3. 3.Comparative Biosciences Department, School of Veterinary MedicineUniversity of Wisconsin-MadisonMadisonUSA
  4. 4.Department of Anatomy and NeuroscienceUniversity of MelbourneMelbourneAustralia

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