Skip to main content
SpringerLink
Log in

Reduced sympathetic neurite outgrowth on uterine tissue sections from rats treated with estrogen

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

In order to evaluate the contribution of substrate-bound factors to the extent and patterning of the sympathetic innervation of rat uterus following estrogen treatment, superior cervical ganglion explants from neonatal and adult ovariectomized rats were cultured on tissue sections of fresh frozen uterus from adult ovariectomized rats treated with estrogen or a vehicle. The main findings were: (1) neurite growth was greatly influenced by histological features of the underlying section; (2) on myometrial sections, neurites followed the orientation of the main axis of the longitudinally sectioned muscle cells; (3) neurites showed limited growth on transversally sectioned smooth muscle; (4) neuritic patterning was unaffected by a reduction in migrating ganglionic non-neuronal cells; (5) neurite outgrowth, but not non-neural cell migration, was markedly reduced on myometrial sections from rats treated with estrogen. These results suggest that adult myometrium continues to provide signals allowing the organotypic patterning and growth of sympathetic axons, that estrogen treatment modifies myometrial substrate properties so that it is less supportive for sympathetic neurite growth, and that adult sympathetic neurons retain their ability to recognize substrate-bound cues present in the myometrium. On endometrial sections, neurites formed radially symmetric halos, which were reduced in size on estrogen-treated endometrial substrates. Thus, changes in the neuritogenic capacity of the uterus underlie plasticity in uterine sympathetic nerves, and alterations in substrate-bound factors contribute to the diminished receptivity of the estrogenized uterus to its sympathetic innervation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  • Adolfsson PI, Huang I, Berg G, Svensson SPS (2000) Changes in β2-adrenoceptors expression and in adenylyl cyclase and phosphodiesterase activity in human uterine leiomyomas. Mol Hum Reprod 6:835–842

    Article  CAS  PubMed  Google Scholar 

  • Akira S, Iwasaki N, Ichikawa M, Mine K, Kuwabara Y, Takeshita T, Tajima H (2009) Successful long-term management of adenomyosis associated with deep thrombosis by low-dose gonadotropin-releasing hormone agonist therapy. Clin Exp Obstet Gynecol 36:123–125

    CAS  PubMed  Google Scholar 

  • Anaf V, Simon P, Nakadi I, Fay I, Simonart T, Buxant F, Noel JC (2002) Hyperalgesia, nerve infiltration and nerve growth factor expression in deep adenomyotic nodules, peritoneal and ovarian endometriosis. Hum Reprod 17:1895–1900

    Article  PubMed  Google Scholar 

  • Anton E, Weskamp G, Reichardt LF, Matthew WD (1994) Nerve growth factor and its low–affinity receptor promotes Schwann cell migration. Proc Natl Acad Sci USA 91:2795–2799

    Article  CAS  PubMed  Google Scholar 

  • Berkley KJ, Dmitrieva N, Curtis KS, Papka RE (2004) Innervation of ectopic endometrium in a rat model of endometriosis. Proc Natl Acad Sci USA 101:11094–11098

    Article  CAS  PubMed  Google Scholar 

  • Berkley KJ, McAllister SL, Accius BE, Winnard KP (2007) Endometriosis-induced vaginal hyperalgesia in the rat: effect of estropause, ovariectomy, and estradiol replacement. Pain 132:S150–S159

    Article  CAS  PubMed  Google Scholar 

  • Bianchimano P, Richeri A, Blasina F, Vaamonde L, Crutcher K, Brauer MM (2008) Substrate contributes to patterning of uterine sympathetic nerves (abstract). 4th International Meeting of the Latin American Society of Developmental Biology. Buenos Aires, Argentine, p 25

  • Borel JP (1991) Uterine collagens. General review. Rev Fr Gynecol Obstet 86:715–722

    CAS  PubMed  Google Scholar 

  • Brauer MM (2008) Cellular and molecular mechanisms underlying plasticity in uterine sympathetic nerves. Auton Neurosci 140:1–16

    Article  CAS  PubMed  Google Scholar 

  • Brauer MM, Burnstock G, Thrasivoulou C, Cowen T (1998) In oculo transplants of myometrium from postpartum guinea pigs fail to support sympathetic reinnervation. J Anat 193:509–517

    Article  PubMed  Google Scholar 

  • Brauer MM, Chávez R, Llodrá J, Richeri A, Scorza C (2000) Effects of chronic oestrogen treatment are not selective for uterine noradrenaline-containing sympathetic nerves: a transplantation study. J Anat 196:347–355

    Article  CAS  PubMed  Google Scholar 

  • Brauer MM, Chávez-Genaro R, Richeri A, Viettro L, Frias AI, Burnstock G, Cowen T (2002) The oestrogenized rat myometrium inhibits organotypic sympathetic reinneravtion. Auton Neurosci 101:13–22

    Article  CAS  PubMed  Google Scholar 

  • Chalar C, Richeri A, Viettro L, Chávez-Genaro R, Bianchimano P, Marmol NM, Crutcher K, Burnstock G, Cowen T, Brauer MM (2003) Plasticity in developing rat uterine sensory nerves: the role of NGF and TrkA. Cell Tissue Res 314:191–205

    Article  CAS  PubMed  Google Scholar 

  • Chávez-Genaro R, Crutcher K, Viettro L, Richeri A, Coirolo N, Burnstock G, Cowen T, Brauer MM (2002) Differential effects of estrogen on developing and mature uterine sympathetic nerves. Cell Tissue Res 308:61–73

    Article  PubMed  Google Scholar 

  • Ceballos D, Navarro X, Dubey N, Wendelschafer-Crabb G, Kennedy WR, Tranquillo RT (1999) Magnetically aligned collagen gel filling a collagen nerve guide improves peripheral nerve regeneration. Exp Neurol 158:290–300

    Article  CAS  PubMed  Google Scholar 

  • Cidadão AJ, Thorsteinsdóttir S, David-Ferreira JF (1990) Immunocytochemical study of tissue distribution and hormonal control of chondroitin-, dermatan- and keratan sulfates from rodent uterus. Eur J Cell Biol 52:105–116

    PubMed  Google Scholar 

  • Covault J, Cunningham JM, Sanes JR (1987) Neurite outgrowth on cryostat tissue sections of innervated and denervated skeletal muscle. J Cell Biol 105:2479–2488

    Article  CAS  PubMed  Google Scholar 

  • Crutcher KA (1993) Tissue sections as culture substrates: overview and critique. Hippocampus 3:157–164

    PubMed  Google Scholar 

  • Crutcher KA, Jayasinghe C, Yun YH, Shanov VN (2009) Progress in the use of aligned carbon nanotubes to support neuronal attachment and directional neurite growth. In: Schulz MJ, Shanov VN (ed) Nanomedicine materials, devices, and systems. Artech House, Boston London, pp 187–207

    Google Scholar 

  • Damon DH (2006) Vascular endothelial-derived semaphorin 3 inhibits sympathetic axon growth. Am J Physiol Heart Physiol 290:H1220–H1225

    Article  CAS  Google Scholar 

  • Dubey N, Letourneau PC, Tranquillo RT (1999) Guided neurite elongation and Schwann cell invasion into magnetically aligned collagen in simulated peripheral nerve regeneration. Exp Neurol 158:338–350

    Article  CAS  PubMed  Google Scholar 

  • Gavazzi I, Boyle KS, Cowen T (1996) Extracellular matrix molecules influence innervation density in rat cerebral blood vessels. Brain Res 734:167–174

    Article  CAS  PubMed  Google Scholar 

  • Hill CE, Chamley JH, Burnstock G (1974) Cell surfaces and fibre relationships in sympathetic ganglion cultures: a scanning electron-microscopic study. J Cell Sci 14:657–669

    CAS  PubMed  Google Scholar 

  • Hou ST, Jiang SX, Smith RA (2008) Permissive and repulsive cues and signaling pathways of axonal outgrowth and regeneration. Int Rev Cell Mol Biol 267:125–181

    Article  CAS  PubMed  Google Scholar 

  • Houdeau E, Rousseau A, Meusnier C, Prud’homme MJ, Rousseau JP (1998) Sympathetic innervation of the upper and lower regions of the uterus and cervix in the rat has different origins and routes. J Comp Neurol 399:403–412

    Article  CAS  PubMed  Google Scholar 

  • Hynds DL, Snow DM (2001) Fibronectin and laminin elicit differential behaviors from SH-SY5Y growth cones contacting inhibitory chondroitin sulfate proteoglycans. J Neurosci Res 66:630–642

    Article  CAS  PubMed  Google Scholar 

  • Krizsan-Agbas D, Smith PG (2002) Estrogen modulates myometrium-induced sympathetic neurite formation through actions on target and ganglion. Neuroscience 114:339–347

    Article  CAS  PubMed  Google Scholar 

  • Krizsan-Agbas D, Pedchenko T, Hassan W, Smith PG (2003) Estrogen regulates sympathetic neurite outgrowth by modulating brain-derived neurotrophic factor synthesis and release by the rodent uterus. Eur J Neurosci 18:2760–2768

    Article  CAS  PubMed  Google Scholar 

  • Krizsan-Agbas D, Pedchenko T, Smith PG (2008) Neurotrimin is an estrogen-regulated determinant of peripheral sympathetic innervation. J Neurosci Res 86:3086–3095

    Article  CAS  PubMed  Google Scholar 

  • Li GN, Liu J, Hoffman-Kim D (2008) Multi-molecular gradients of permissive and inhibitory cues direct neurite outgrowth. Ann Biomed Eng 36:889–904

    Article  PubMed  Google Scholar 

  • Lobos E, Gebhardt C, Kluge A, Spanel-Borowski K (2005) Expression of nerve growth factor (NGF) isoforms in the rat uterus during pregnancy: accumulation of precursor proNGF. Endocrinology 146:1922–1929

    Article  CAS  PubMed  Google Scholar 

  • Mark K von der, Park J, Bauer S, Schmuki P (2010) Nanoscale engineering of biomimetic surfaces: cues from the extracellular matrix. Cell Tissue Res 339:131–153

    Article  PubMed  Google Scholar 

  • Martínez G, Silveira P, Bianchimano P, Richeri A, Brauer MM (2009) El estrógeno reduce la capacidad neuritogénica del sustrato uterino. Papel de la geometría del colágeno (abstract). Sextas Jornadas de la Sociedad de Bioquímica y Biología Molecular, Montevideo, Uruguay, p 75

  • Marzioni D, Tamagnone L, Capparuccia L, Marchini C, Amici A, Todros T, Bischof P, Neidhart S, Grenningloh G, Castellucci M (2004) Restricted innervation of uterus and placenta during pregnancy: evidence for a role of the repelling signal semaphorin 3A. Dev Dyn 231:839–848

    Article  CAS  PubMed  Google Scholar 

  • Medina MG, Lebovic DI (2009) Endometriosis-associated nerve fibers and pain. Acta Obstet Gynecol Scand 88:968–975

    Article  PubMed  Google Scholar 

  • Munakata H, Isemura M, Yosizawa Z (1984) Sulfated proteoglycans synthesized in myometrium of the estrogen-treated rabbit. Tohoku J Exp Med 144:291–297

    Article  CAS  PubMed  Google Scholar 

  • Nishinaka K, Fukuda Y (1991) Changes in extracellular matrix materials in the uterine myometrium of rats during pregnancy and postparturition. Acta Pathol Jpn 41:122–132

    CAS  PubMed  Google Scholar 

  • Orike N, Thrasivoulou C, Wrigley A, Cowen T (2001) Differential regulation of survival and growth in adult sympathetic neurons: an in vitro study of neurotrophin responsiveness. J Neurobiol 47:295–305

    Article  CAS  PubMed  Google Scholar 

  • Papka RE, Cotton JP, Traurig HH (1985) Comparative distribution of neuropeptide tyrosine-, vasoactive intestinal polypeptide-, substance P-immunoreactive, acetylcholinesterase-positive and noradrenergic nerves in the reproductive tract of the female rat. Cell Tissue Res 242:475–490

    Article  CAS  PubMed  Google Scholar 

  • Pettigrew DB, Crutcher KA (1999) White matter of the CNS supports or inhibits neurite outgrowth in vitro depending on geometry. J Neurosci 19:8358–8366

    CAS  PubMed  Google Scholar 

  • Pettigrew DB, Crutcher KA (2001) Myelin contributes to the parallel orientation of axonal growth on white matter in vitro. BMC Neurosci 2:9

    Article  CAS  PubMed  Google Scholar 

  • Pettigrew DB, Shockely KP, Crutcher KA (2001) Disruption of spinal cord white matter and sciatic nerve geometry inhibits axonal growth in vitro in the absence of glial scarring. BMC Neurosci 2:8

    Article  CAS  PubMed  Google Scholar 

  • Quinn M (2007a) Uterine innervation in adenomyosis. J Obstet Gynaecol 27:287–291

    Article  CAS  PubMed  Google Scholar 

  • Quinn M (2007b) Uterine innervation in fibroids: a qualitative study. J Obstet Gynaecol 27:489–492

    Article  CAS  PubMed  Google Scholar 

  • Richeri A, Bianchimano P, Mármol NM, Viettro L, Cowen T, Brauer MM (2005) Plasticity in rat uterine sympathetic nerves: the role of TrkA and p75 nerve growth factor receptors. J Anat 207:125–134

    Article  CAS  PubMed  Google Scholar 

  • Richeri A, Chalar C, Bianchimano P, Greif G, Brauer MM (2008) Estrogen regulation of semaphorin expression in the rat uterus (abstract). EMBO Workshop Semaphorin Function and Mechamisms of Action. Abbaye des Vaulx de Cernay, France

  • Ross R, Klebanoff SJ (1971) The smooth muscle cell. 1. In vivo synthesis of connective tissue proteins. J Cell Biol 50:159–171

    Article  CAS  PubMed  Google Scholar 

  • Rydhström H, Walles B, Owman C (1989) Myometrial norepinephrine in human pregnancy—abnormally elevated levels at term related to various disorders leading to Cesarean section. Am J Reprod Med 34:901–904

    Google Scholar 

  • Shynlova O, Mitchell JA, Tsampalieros A, Langille AL, Lye SJ (2004) Progesterone and gravidity differentially regulate expression of extracellular matrix components in the pregnant rat myometrium. Biol Reprod 70:986–992

    Article  CAS  PubMed  Google Scholar 

  • Slater M, Murphy CR (1999) Chondroitin sulfate and heparan sulfate proteoglycans are sequentially expressed in the uterine extracellular matrix during early pregnancy in the rat. Matrix Biol 18:125–131

    Article  CAS  PubMed  Google Scholar 

  • Snow DM, Smith JD, Gurwell JA (2002) Binding characteristics of chondroitin sulphate proteoglycans and laminin-1, and correlative neurite outgrowth behaviors in standard tissue culture assays. J Neurobiol 51:285–301

    Article  CAS  PubMed  Google Scholar 

  • Stein K, Ascher-Walsh C (2009) A comprehensive approach to the treatment of uterine leiomyomata. Mt Sinai J Med 76:546–556

    Article  PubMed  Google Scholar 

  • Stewart EA, Floor AE, Jain P, Nowak RA (1995) Increased expression of messenger RNA for collagen type I, collagen type II, and fibronectin in myometrium of pregnancy. Obstet Gynecol 86:417–422

    Article  CAS  PubMed  Google Scholar 

  • Tokushige N, Markham R, Russell P, Fraser IS (2006) Nerve fibres in peritoneal endometriosis. Hum Reprod 21:3001–3007

    Article  CAS  PubMed  Google Scholar 

  • Tokushige N, Markham R, Russell P, Fraser IS (2007) Different types of small nerve fibers in eutopic endometrium and myometrium in women with endometriosis. Fertil Steril 88:795–803

    Article  PubMed  Google Scholar 

  • Tuttle R, Matthew WD (1991) An in vitro assay for neurite growth using cryostat sections of nervous tissue as substratum. J Neurosci Methods 39:193–202

    Article  CAS  PubMed  Google Scholar 

  • Vera PL, Haase EB, Schramm LP (1997) Origins of the sympathetic innervation of the cervical end of the uterus in the rat. Brain Res 747:140–143

    Article  CAS  PubMed  Google Scholar 

  • Vercellini P, Somigliana E, Viganò P, Abbiati A, Barbara G, Crosignani PG (2009) Endometriosis: current therapies and new pharmacological developments. Drugs 69:649–675

    Article  CAS  PubMed  Google Scholar 

  • Zoubina EV, Smith PG (2001) Sympathetic hyperinnervation of the uterus in the estrogen receptor alpha knock-out mouse. Neuroscience 103:237–244

    Article  CAS  PubMed  Google Scholar 

  • Zoubina EV, Fan Q, Smith PG (1998) Variation in uterine innervation during the estrous cycle. J Comp Neurol 397:561–571

    Article  CAS  PubMed  Google Scholar 

  • Zoubina EV, Mize AL, Alper RH, Smith PG (2001) Acute and chronic oestrogen supplementation decreases uterine sympathetic innervation in ovariectomised adult virgin rats. Histol Histopathol 16:989–996

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Special thanks are extended to Dr. Fernanda Blasina, Lucía Vaamonde, and the staff of the Neurochemistry Department (IIBCE) for their technical assistance and valuable suggestions. The technical support and electron micrographs provided by Gaby Martínez and Patricia Silveira are greatly appreciated. Critical reading of the manuscript by Professor Timothy Cowen and Dr. David Pettigrew is gratefully acknowledged.

Author information

Authors and Affiliations

  1. Laboratory of Cell Biology, Instituto de Investigaciones Biológicas Clemente Estable, Avenida Italia 3318, Montevideo, 11600, Uruguay

    Analía Richeri, Paola Bianchimano & M. Mónica Brauer

  2. Department of Neurosurgery, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH, 45267-0515, USA

    Keith A. Crutcher

Authors
  1. Analía Richeri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paola Bianchimano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Keith A. Crutcher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Mónica Brauer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Mónica Brauer.

Additional information

Analía Richeri and Paola Bianchimano contributed equally to this work and are postgraduate students of PEDECIBA, Uruguay. This work was supported by a Fogarty International Research Collaboration Award (FIRCA, NIH, grant no. TW007213).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Richeri, A., Bianchimano, P., Crutcher, K.A. et al. Reduced sympathetic neurite outgrowth on uterine tissue sections from rats treated with estrogen. Cell Tissue Res 340, 287–301 (2010). https://doi.org/10.1007/s00441-010-0956-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-010-0956-x

Keywords

Search

Navigation